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Cultural and Environmental Factors Influence the Performance of Angelonia augustifolia Cultivars

Permanent Link: http://ufdc.ufl.edu/UFE0022406/00001

Material Information

Title: Cultural and Environmental Factors Influence the Performance of Angelonia augustifolia Cultivars
Physical Description: 1 online resource (139 p.)
Language: english
Creator: Boldt, Jennifer
Publisher: University of Florida
Place of Publication: Gainesville, Fla.
Publication Date: 2008

Subjects

Subjects / Keywords: angelonia, angustifolia, flowering, growth, irradiance, plant, regulators, snapdragon, summer, temperature
Environmental Horticulture -- Dissertations, Academic -- UF
Genre: Horticultural Science thesis, M.S.
bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

Abstract: Thirty-one commercial Angelonia angustifolia Benth. cultivars were evaluated for phenotypic differences in growth and flowering. Number of days to first flower and marketability during spring production ranged from 41 to 64 d and 45 to 70 d, respectively. At marketability, plant height ranged from 14 to 64 cm, plant width from 28 to 54 cm, and inflorescence number from 2.3 to 4.2. At 9.5 weeks after pinch, inflorescence number ranged from 2.1 to 29.2. Flower size ranged from 20.1 to 30.7 mm tall and 12.7 to 28.4 mm wide. Cultivars differed in powdery mildew sensitivity. In a sub-sample of eight cultivars, a season x cultivar interaction was significant for all parameters except flower size. Time to first flower decreased and plant height increased from season 1 to 4, but the magnitude differed with cultivar. Eight cultivars planted in a landscape bed at three intervals differed significantly in summer performance. Ten weeks after transplant, the interaction was significant for plant height, inflorescence number, and flower size. Decreased summer flowering was influenced by environmental conditions rather than plant age. Summer flowering was poor in all cultivars except AngelMist Purple Stripe . Cultivars subirrigated with increasing fertilizer concentrations exhibited an interaction for all parameters except final pH and number of days to first flower, which were due to the main effect of fertilizer concentration and cultivar, respectively. SPAD values increased as N concentration increased. EC increased and pH decreased as N concentration increased. Optimal plant size, dry weight, and inflorescence number occurred at different concentrations. Responses to fertilization were not cultivar specific and recommended concentrations are at N levels of 100 mg/L or slightly higher. Cultivars showed varying sensitivities to paclobutrazol, daminozide, and ethephon. All effectively inhibited growth in at least one cultivar, and paclobutrazol drenches inhibited stem elongation in all cultivars. Paclobutrazol drenches of 5 mg/L applied 2 weeks after pinch inhibited stem elongation, but sprays of 100 mg/L applied 2 weeks after pinch and drenches of 8 mg/L applied at visible bud were ineffective. Ethephon inhibited stem elongation, increased lateral branching, delayed flowering, and caused phytotoxicity.
General Note: In the series University of Florida Digital Collections.
General Note: Includes vita.
Bibliography: Includes bibliographical references.
Source of Description: Description based on online resource; title from PDF title page.
Source of Description: This bibliographic record is available under the Creative Commons CC0 public domain dedication. The University of Florida Libraries, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Statement of Responsibility: by Jennifer Boldt.
Thesis: Thesis (M.S.)--University of Florida, 2008.
Local: Adviser: Barrett, James E.

Record Information

Source Institution: UFRGP
Rights Management: Applicable rights reserved.
Classification: lcc - LD1780 2008
System ID: UFE0022406:00001

Permanent Link: http://ufdc.ufl.edu/UFE0022406/00001

Material Information

Title: Cultural and Environmental Factors Influence the Performance of Angelonia augustifolia Cultivars
Physical Description: 1 online resource (139 p.)
Language: english
Creator: Boldt, Jennifer
Publisher: University of Florida
Place of Publication: Gainesville, Fla.
Publication Date: 2008

Subjects

Subjects / Keywords: angelonia, angustifolia, flowering, growth, irradiance, plant, regulators, snapdragon, summer, temperature
Environmental Horticulture -- Dissertations, Academic -- UF
Genre: Horticultural Science thesis, M.S.
bibliography   ( marcgt )
theses   ( marcgt )
government publication (state, provincial, terriorial, dependent)   ( marcgt )
born-digital   ( sobekcm )
Electronic Thesis or Dissertation

Notes

Abstract: Thirty-one commercial Angelonia angustifolia Benth. cultivars were evaluated for phenotypic differences in growth and flowering. Number of days to first flower and marketability during spring production ranged from 41 to 64 d and 45 to 70 d, respectively. At marketability, plant height ranged from 14 to 64 cm, plant width from 28 to 54 cm, and inflorescence number from 2.3 to 4.2. At 9.5 weeks after pinch, inflorescence number ranged from 2.1 to 29.2. Flower size ranged from 20.1 to 30.7 mm tall and 12.7 to 28.4 mm wide. Cultivars differed in powdery mildew sensitivity. In a sub-sample of eight cultivars, a season x cultivar interaction was significant for all parameters except flower size. Time to first flower decreased and plant height increased from season 1 to 4, but the magnitude differed with cultivar. Eight cultivars planted in a landscape bed at three intervals differed significantly in summer performance. Ten weeks after transplant, the interaction was significant for plant height, inflorescence number, and flower size. Decreased summer flowering was influenced by environmental conditions rather than plant age. Summer flowering was poor in all cultivars except AngelMist Purple Stripe . Cultivars subirrigated with increasing fertilizer concentrations exhibited an interaction for all parameters except final pH and number of days to first flower, which were due to the main effect of fertilizer concentration and cultivar, respectively. SPAD values increased as N concentration increased. EC increased and pH decreased as N concentration increased. Optimal plant size, dry weight, and inflorescence number occurred at different concentrations. Responses to fertilization were not cultivar specific and recommended concentrations are at N levels of 100 mg/L or slightly higher. Cultivars showed varying sensitivities to paclobutrazol, daminozide, and ethephon. All effectively inhibited growth in at least one cultivar, and paclobutrazol drenches inhibited stem elongation in all cultivars. Paclobutrazol drenches of 5 mg/L applied 2 weeks after pinch inhibited stem elongation, but sprays of 100 mg/L applied 2 weeks after pinch and drenches of 8 mg/L applied at visible bud were ineffective. Ethephon inhibited stem elongation, increased lateral branching, delayed flowering, and caused phytotoxicity.
General Note: In the series University of Florida Digital Collections.
General Note: Includes vita.
Bibliography: Includes bibliographical references.
Source of Description: Description based on online resource; title from PDF title page.
Source of Description: This bibliographic record is available under the Creative Commons CC0 public domain dedication. The University of Florida Libraries, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Statement of Responsibility: by Jennifer Boldt.
Thesis: Thesis (M.S.)--University of Florida, 2008.
Local: Adviser: Barrett, James E.

Record Information

Source Institution: UFRGP
Rights Management: Applicable rights reserved.
Classification: lcc - LD1780 2008
System ID: UFE0022406:00001


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CULTURAL AND ENVIRONMENTAL FACTORS INFLUENCE THE PERFORMANCE OF
Angelonia angustifolia CULTIVARS




















By

JENNIFER KAY BOLDT


A THESIS PRESENTED TO THE GRADUATE SCHOOL
OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT
OF THE REQUIREMENTS FOR THE DEGREE OF
MASTER OF SCIENCE

UNIVERSITY OF FLORIDA

2008


































2008 Jennifer Kay Boldt





























To Mom and Dad









ACKNOWLEDGMENTS

I would like to thank my advisor and committee chair, Dr. James Barrett, for all of his

guidance and support during my years as an undergraduate and graduate student at the University

of Florida. Without his nudging, I may not have decided to pursue a master's program. During

the past two and a half years, he listened to all of my ideas (both the good and the bad), provided

unwavering support for my research, and kept me well-fed during long days of data collection

and trial garden planting. I would also like to thank my committee members, Drs. James Gibson,

Rosanna Freyre, and Kenneth Quesenberry, for their support and guidance.

I thank Ms. Carolyn Bartuska for patiently answering all of my many statistical

questions. More importantly, she managed to survive working in the same office as my twin

sister and me, which was no easy task when we both got stressed! Weekly lunch outings

somehow managed to keep us all sane. Thanks also go to Bob Weidman for taking care of my

research plants when I was out of town; without alive plants, I would not have had any results.

Much appreciation is given to my twin sister Jessica for continually offering to help me

collect data in a hot, humid greenhouse when no one else was available to help. She was always

the behind-the-scenes person making sure everything was taken care of so that I could finish

experiments and write this thesis.

Finally, I would like to thank my parents for all of their support and guidance. They

provided me with my first glimpse into the world of floriculture, taught me the importance of

hard work and finishing a task, and patiently listened to all of my joys and frustrations.









TABLE OF CONTENTS

page

A CK N O W LED G M EN T S ................................................................. ........... ............. .....

L IST O F T A B L E S ......................................... ..............................8

LIST OF FIGURES .................................. .. .... ...... ........... ...... 10

LIST OF FIGURES .................................. .. .... ...... ................... 10

A B S T R A C T ......... ....................... ............................................................ 12

CHAPTER

1 L IT E R A TU R E R E V IE W ............................................................................... .................. 14

In tro d u c tio n ............................................ .......................................................................... 1 4
A n g e lo n ia ....................... .........................................................................................................1 5
T ax o n o m y ........................................................................................................1 5
B otany .......................................................................................................... ............ 15
P ollination and R production .................................................................................... 16
D described Species.................................................17
C u ltiv a tio n .......................................................................................................................1 8
Commercial Cultivars..............................................19
F low erin g ..................................... ........................................................19
Irradiance Influences Time to Flower ....................................................... 21
Temperature Influences Time to Flower ...................... ........ 22
Temperature x Irradiance Interactions Influence Time to Flower...............................23
F low erin g in A n g elon ia ............................................................................................. 2 4
Factors Influencing Continued Flowering ................................ ...............................25
P la n t G ro w th ..................................................................................................................... 2 7
T e m p e ratu re ............................................................................................................... 2 7
Irra d ia n c e .............................. .............................................................2 8
Temperature x Irradiance Interactions ..................................... .....................29
F e rtiliz e rs .............. .... ...............................................................3 0
P lan t G row th ............................................... .........................................3 0
Use of a Hand-held Meter to Measure Chlorophyll Content ........................................ 31
P lant G row th R egu lators................................................................................................... 3 3
In tro d u ctio n ..............................................................................3 3
P aclo b u traz o l .............................................................................3 4
D am inozide ............................................. 37
E th e p h o n ................................................................................3 8
O bje ctiv e s ................... ...................4...................1..........









2 Angelonia angustifolia CULTIVAR SCREEN ............................. .................... 44

Intro du action ......... ............................................................................... 4 4
Materials and Methods ................................. ............ ...... .............. 44
Experiment 2-1 Cultivar Screen of 31 Commercial Cultivars .....................................44
Experiment 2-2 Cultivar Comparison of Seven Cultivars across Multiple Seasons.......46
Results and Discussion ..................................... ................. ........ ..... 47
E x p erim ent 2 -1 ................................................................4 7
D ay s to first flow er............ ................................................................ ....... . ...... 47
D ay s to m marketability ................................................... .............. ... ............ 48
Plant height and width at marketability........................ ........................ 49
Inflorescence number at marketability ................. ............................................. 50
Inflorescence num ber at 9.5 w eeks ........................................ ....... ............... 51
F lo w e r siz e ............................................................................................................... 5 1
Pow dery m ildew index .............................................. ........ ......................... 52
E x p erim en t 2 -2 ................................................................................5 3
D ay s to first flow er............ ................................................................ ....... . ...... 53
P lant height...................................................................................... 55
F lo w er siz e .........................................................................5 6
Pow dery m ildew susceptibility ........................................ .......................... 57
C onclu sions.......... ............................... ................................................57

3 PLANT DATE x CULTIVAR INTERACTIONS INFLUENCE SUMMER
LANDSCAPE PERFORMANCE OF Angelonia angustifolia............... ... .............. 69

Introduction ............ ........ .................................69
M materials and M methods ................. .................................... .. ........ .. .............69
Results and Discussion ................. ........ ................................. .. 71
10-W eek D ata ........................................................................................... 71
P lan t h eig h t.....................................................7 1
Infl orescence num ber ................................................................... .... ..................72
F lo w e r h e ig h t...................................................................................................... 7 3
Flow er Ratings by Plant Age........................................................................ 74
Flow er R atings by Evaluation D ate........................................................ ............... 75
C onclu sions.......... ............................... ................................................77

4 CULTIVAR BY FERTILIZER INTERACTIONS AFFECT GROWTH OF Angelonia
a ng u stif o lia .........................................................................8 3

Introduction ............ ........ .................................83
M materials and M methods ...................................... .. .......... ....... ...... 84
Results and Discussion ....................... .. .. ..... .. .... ........ ........ .. 85
Substrate pH and Electrical Conductivity ............................................ ............... 85
L eaf G reenn ess ................................................................87
P la n t G ro w th .............................................................................................................. 8 9
F lo w e rin g ........................................................ ... .............................................9 0
C onclu sions.......... ..........................................................9 1


6









5 Angelonia angustifolia CULTIVARS DIFFER IN RESPONSE TO
PACLOBUTRAZOL, DAMINOZIDE, AND ETHEPHON............................102

Introduction ..................................................................102
M materials and M methods ........................ .................. ...... ... .......... ............... 103
Experiment 5-1 Paclobutrazol, Daminozide, and Ethephon Response Curves............103
Experiment 5-2 Daminozide....................... ........ ........................ 104
Experiment 5-3 Paclobutrazol Spray and Drench Applications.................................. 105
E xperim ent 5-4 E thephon ................................................................. .................... 105
R results and D discussion ..................................... ....................... .... .............. 107
E x p erim en t 5 -1 .............................................................................................................. 10 7
Paclobutrazol ......................................... .................. .. .... ........ 107
D am in ozide ....................................................... 10 8
E th e p h o n ........................................................................................................... 1 0 9
E x p erim en t 5 -2 .............................................................................................................. 10 9
E x p erim en t 5 -3 ............................................................ ................................... 1 1 1
E x p e rim e n t 5 -4 ................................................................... ...................................1 12
Phytotoxicity ......................................... .................. ... .... ........ 112
Stem elongation ............................................................................................. 113
Lateral num ber ................................................................... ... ......... 114
F lo w e rin g .....................................................................................................1 14
C onclu sions.......... .........................................................115

6 C O N C L U S IO N S ............................................................................................................ 12 5

L IST O F R E F E R E N C E S ...................................................................................................129

BIOGRAPHICAL SKETCH .........................................................................139
























7









LIST OF TABLES


Table page

1-1 Current taxonomic classification ofAngelonia angustifolia. .........................................42

1-2 Com m ercially available angelonia series...................................... ......................... 42

1-3 Cultivar names of angelonia used in experiments. ................................. .................43

2-1 Plant growth and flowering characteristics for seven Angelonia angustifolia cultivars
u sed in E xpt. 2-2 ........................................................ ................. 6 1

2-2 Temperature and light data for Expt. 2-2............... .......... ......... .................... 62

2-3 Angelonia angustifolia flowering data for 31 cultivars (Expt. 2-1)..............................63

2-4 Plant data for 31 cultivars of Angelonia angustifolia collected at marketability (Expt.
2-1).......................................................... 64

2-5 Angelonia angustifolia plant data for 31 cultivars collected 9.5 weeks after pinch
(E xpt. 2-1). ..................................................................................65

2-6 Split plot analysis of variance for seven Angelonia angustifolia cultivars grown at
four different seasons from January to July 2007 (Expt. 2-2). ........................................66

2-7 Number of days to first flower in seven Angelonia angustifolia cultivars across four
seasons (E xpt. 2-2) ........................................................................ 66

2-8 Plant height (cm) at marketability in seven Angelonia angustifolia cultivars across
four seasons (Expt. 2-2)................... .......................... ........... 67

2-9 Flower height (mm) in six Angelonia angustifolia cultivars across four seasons
(E xpt. 2-2). .................................................................................67

2-10 Powdery mildew index in seven Angelonia angustifolia cultivars across four seasons
(E xpt. 2-2). .................................................................................68

3-1 Monthly temperature and DLI data for Gainesville, FL for summer 2007. ....................80

3-2 Split-plot analysis of variance for Angelonia angustifolia growth and flowering data
collected at 10 weeks after planting for three plant dates ............................................80

3-3 Mean separation for plant height (cm) of eight Angelonia angustifolia cultivars
collected at 10 weeks after transplant for three plant dates. ............................................81

3-4 Mean separation for number of inflorescences per plant of eight Angelonia
angustifolia cultivars collected at 10 weeks after transplant for three plant dates. ...........81









3-5 Mean separation for flower height (mm) of eight Angelonia angustifolia cultivars
collected at 10 weeks after transplant for three plant dates. ............................................81

3-6 Split-plot analysis of variance for Angelonia angustifolia flower ratings by plant age
(w weeks after transplant)......... ......................................................... ........ .. 82

3-7 Split-plot analysis of variance for Angelonia angustifolia flower ratings by
evaluation date. ............................................................................82

4-1 Analysis of variance for Angelonia angustifolia cultivars in response to fertilization......98

4-2 Regression equations for pH and EC measurements collected at 6 weeks after
treatm ent ........................................................ .................................98

4-3 Regression equations for SPAD readings collected at 6 weeks after treatment ...............99

4-4 Regression equations for plant size, dry weight, and number of inflorescences at 6
weeks after treatment .............................................. ......... 100

4-5 Number of days to first open flower in Angelonia angustifolia. ..................................... 101

5-1 Regression equations for stem elongation for Angelonia angustifolia cultivars in
response to PGR concentration (Expt. 5-1). ....................................... ............... 122

5-2 Stem elongation (cm) in Angelonia angustifolia four weeks after application of
dam inozide (E xpt. 5-2). .......................... ...... ..................................... .. .... .. 123

5-3 Stem elongation (cm) in Angelonia angustifolia in response to paclobutrazol
applications (E xpt. 5-3) ................................................................... 123

5-4 Analysis of variance for Angelonia angustifolia treated with ethephon (Expt. 5-4). ......123

5-5 Mean separation for severity of phytotoxicity symptoms in Angelonia angustifolia
one week after treatment with ethephon (Expt. 5-4)...............................................124

5-6 Regression equations for Angelonia angustifolia cultivars treated with ethephon
(E xpt. 5-4). ...............................................................................124

5-7 Mean separation for number of laterals in Angelonia angustifolia 4 weeks after
ethephon treatm ent (Expt. 5-4). .............................................. ............................. 124









LIST OF FIGURES


Figure page

2-1 Range of plant height present in commercial Angelonia angustifolia cultivars ................59

2-2 Examples of the range on inflorescence number present in Angelonia angustifolia
cultivars 9.5 weeks after pinching ................................. .....................................59

2-3 Range of flower size present in commercial Angelonia angustifolia cultivars .................60

2-4 Comparison between inflorescences in Angelonia angustifolia 'Angelface White'
and 'Serena W white' .........................................................................60

3-1 Flower ratings of Angelonia angustifolia graphed by weeks after transplant (WAT).......78

3-2 Flower ratings of Angelonia angustifolia graphed by date of data collection. ................79

4-1 Effect of increasing fertilization with 20.0N-4.4P-16.6K on substrate pH in
Angelonia angustifolia 6 weeks after start of treatments.................... .. ............... 93

4-2 Effect of increasing fertilization with 20.0N-4.4P-16.6K on substrate electrical
conductivity in Angelonia angustifolia 6 weeks after start of treatments........................93

4-3 Effect of increasing fertilization with 20.0N-4.4P-16.6K on lower leaf SPAD values
in Angelonia angustifolia 6 weeks after start of treatments.................. ............... 94

4-4 Effect of increasing fertilization with 20.0N-4.4P-16.6K on upper leaf SPAD values
in Angelonia angustifolia 6 weeks after start of treatments.................. ............... 94

4-5 Response of Angelonia angustifolia 'AngelMist Dark Lavender' to increasing rates
of fertilization with 20.0N-4.4P-16.6K.................. .. ......... ............... ............... 95

4-6 Effect of increasing fertilization with 20.0N-4.4P-16.6K on plant size in Angelonia
angustifolia 6 weeks after start of treatments ............. ............................... .............. 96

4-7 Effect of increasing fertilization with 20.0N-4.4P-16.6K on plant dry weight in
Angelonia angustifolia 6 weeks after start of treatments.................... .. ............... 96

4-8 Effect of increasing fertilization with 20.0N-4.4P-16.6K on number of inflorescences
in Angelonia angustifolia 6 weeks after start of treatments.................. ............... 97

5-1 Effect of plant growth regulators on stem elongation in Angelonia angustifolia 23
days after treatment ........... .... .......... ... ................ ........ .... 17

5-2 Examples of the response of Angelonia angustifolia cultivars to increasing
concentrations of paclobutrazol (mg-L1) (Expt. 5-1) ......................... ..................... 118









5-3 Examples of the response of Angelonia angustifolia cultivars to increasing
concentrations of daminozide (mg-L1) (Expt. 5-1) ....................... .................. 119

5-4 Examples of the response of Angelonia angustifolia cultivars to increasing
concentrations of ethephon (mg-L1) (Expt. 5-1) ................................................. 120

5-5 Stem elongation in Angelonia angustifolia cultivars treated with ethephon (Expt.
5 -4 ) ................... ............................................................................ 12 1

5-6 Days to first open flower in Angelonia angustifolia cultivars treated with ethephon
(E x p t. 5 -4 ) ........................................................................ ................ 12 1









Abstract of Thesis Presented to the Graduate School
of the University of Florida in Partial Fulfillment of the
Requirements for the Degree of Master of Science

CULTURAL AND ENVIRONMENTAL FACTORS INFLUENCE THE PERFORMANCE OF
ANGELONIA ANGUSTIFOLIA CULTIVARS

By

Jennifer Kay Boldt

August 2008

Chair: James Barrett
Major: Horticultural Sciences

Thirty-one commercial Angelonia angustifolia Benth. cultivars were evaluated for

phenotypic differences in growth and flowering. Number of days to first flower and

marketability during spring production ranged from 41 to 64 d and 45 to 70 d, respectively. At

marketability, plant height ranged from 14 to 64 cm, plant width from 28 to 54 cm, and

inflorescence number from 2.3 to 4.2. At 9.5 weeks after pinch, inflorescence number ranged

from 2.1 to 29.2. Flower size ranged from 20.1 to 30.7 mm tall and 12.7 to 28.4 mm wide.

Cultivars differed in powdery mildew sensitivity. In a sub-sample of eight cultivars, a season x

cultivar interaction was significant for all parameters except flower size. Time to first flower

decreased and plant height increased from season 1 to 4, but the magnitude differed with

cultivar.

Eight cultivars planted in a landscape bed at three intervals differed significantly in

summer performance. Ten weeks after transplant, the interaction was significant for plant

height, inflorescence number, and flower size. Decreased summer flowering was influenced by

environmental conditions rather than plant age. Summer flowering was poor in all cultivars

except 'AngelMist Purple Stripe'.









Cultivars subirrigated with increasing fertilizer concentrations exhibited an interaction for

all parameters except final pH and number of days to first flower, which were due to the main

effect of fertilizer concentration and cultivar, respectively. SPAD values increased as N

concentration increased. EC increased and pH decreased as N concentration increased. Optimal

plant size, dry weight, and inflorescence number occurred at different concentrations. Responses

to fertilization were not cultivar specific and recommended concentrations are at N levels of 100

mg/L or slightly higher.

Cultivars showed varying sensitivities to paclobutrazol, daminozide, and ethephon. All

effectively inhibited growth in at least one cultivar, and paclobutrazol drenches inhibited stem

elongation in all cultivars. Paclobutrazol drenches of 5 mg/L applied 2 weeks after pinch

inhibited stem elongation, but sprays of 100 mg/L applied 2 weeks after pinch and drenches of 8

mg/L applied at visible bud were ineffective. Ethephon inhibited stem elongation, increased

lateral branching, delayed flowering, and caused phytotoxicity.









CHAPTER 1
LITERATURE REVIEW

Introduction

The floriculture industry in the late 1980s saw a rise in popularity of new, vegetatively-

propagated annual crops. Consumer interest in new crops and a shift towards container

gardening has helped vegetative annuals become an important product for the industry. In a

15-state survey of growers having at least $10,000 in sales, their total crop value in 2007 was

$4.1 billion and bedding plants accounted for $1.76 billion of the total (NASS, 2008).

Approximately 50% of the bedding plants sold were classified as "other" and not as one of the

staple landscape crops: begonias, geraniums, impatiens, marigolds, pansies, petunias, or violas.

This "other" group is a combination of seed and vegetatively-propagated crops, but it reflects

consumer preference towards selecting a combination of older, stand-by crops and newer, lesser

known crops for gardens.

To catch a consumer's attention in the garden center and entice him or her to purchase a

particular plant, a new crop must have some trait that makes it stand out from the others. Thus,

breeders have focused on developing new cultivars with novel growth habits, flower forms,

and/or flower colors. This is most easily done through the development of vegetatively-

propagated crops due to the quicker selection cycle needed relative to developing uniform seed-

produced cultivars.

Angelonia angustifolia, a vegetatively-propagated or seed-propagated crop, is available

from multiple young plant suppliers. Within the range of cultivars available from each company,

commonly referred to as a series, there are differences in growth habit and flowering time. This

can lead to problems in production, where similar schedules and production requirements are

ideal for large-scale production. Having a range of variability within a species can be both









positive and negative; it provides the consumer with options at retail, but it can make it difficult

for a grower to efficiently produce all of the cultivars. Thus a dilemma arises during new

cultivar selection as to whether or not to release a cultivar and place it within an existing series

even if it is not uniform with the rest of the other cultivars already present in that collection.

Currently, the range of variability present in angelonia cultivars is unknown, both between

different series and within a series. Within the variability present, are there certain traits that are

beneficial to this crop or are there traits that are hindering its success in production and in the

landscape? It will be beneficial to quantify the variability present and determine if current

production recommendations are crop-specific or cultivar-specific.

Angelonia

Taxonomy

Angelonia Humb. & Bonpl., a genus containing approximately 30 species, is native to

Mexico, Central and South America, and the West Indies (Bailey and Bailey, 1976; Huxley,

1992). It originally was placed in Scrophulariaceae (complete taxonomic classification found in

Table 1-1), but taxonomists have reorganized this family in recent years and currently place

Angelonia in Plantaginaceae (Albach et al., 2005). It is diploid with a base chromosome number

of 10 (2n=2x=20) (Steiner, 1996).

Botany

Angelonia species are subshrubs or perennial herbs and range in height from 0.3 to 0.8 m

tall (Bailey and Bailey, 1976). Leaves are opposite in orientation, and may be either glabrous or

pubescent (Bailey and Bailey, 1976; Huxley, 1992). The flowers, borne by themselves in leaf

axils or on a terminal raceme inflorescence (Huxley, 1992), are zygomorphic with a two-lipped

corolla. The upper lip is two-lobed and the lower lip is three-lobed (Griffiths, 1994). The

corolla does not contain a spur, like in other closely-related species, and the corolla tube is highly









reduced or absent entirely (Huxley, 1994). The stigma and four anthers are located on the top of

the "gullet" created by the reduced corolla tube (Vogel and Machado, 1991). The corolla tends

to be blue, lavender, or white-colored (Griffiths, 1994; Vogel and Machado, 1991). Seeds are

produced within a fruit capsule (Bailey and Bailey, 1976), and a distinguishing characteristic of

this genus is its lack of an endosperm in the seed (Steiner, 1996).

Angelonia flowers reward pollinators with oil rather than nectar, first described by Vogel

(1974) to exist in 1260 species, 50 genera, and five families: Iridaceae, Krameriaceae,

Malphigiaceae, Orchidaceae, and Scrophulariaceae. He used the term 'elaiophore' to describe

the glands where the oil is produced. Angelonia flowers have trichome elaiophores, collections

of hundreds of thousands of tiny glands that secrete oil. The oil is absorbed by the tarsal pads of

visiting female bees (Kampny, 1995; Vogel, 1974). Bees, specifically Centris species, are the

common pollinators of Angelonia and use the oil as a food source for larvae (Buchman, 1987;

Vogel and Machado, 1991).

Pollination and Reproduction

The flowers of Angelonia are zygomorphic and designed for insect pollination. Upon

alighting, a bee places its front legs into the flower to collect oil from the pouches. In doing so,

its head comes in contact with the reproductive organs (Vogel and Machado, 1991).

Not surprisingly, due to its close relationship with the oil-collecting bees, the majority of

Angelonia are outcrossing species. Through a combination of field observations and greenhouse

experiments, Machado et al. (2006) determined that among five species of Angelonia observed

from the Caatinga in northeastern Brazil, the four perennial species were self-incompatible and

the annual A. pubescens was self-compatible. Angelonia flowers are protandrous and senesce

five to seven days after opening. The anthers dehisce during the second to fourth days after

flower opening, and the stigma does not become receptive until the fourth or fifth day (Vogel









and Machado, 1991). In its native habitat, flowering occurs from February or March until June

or July and coincides with the rainy season (Vogel and Machado, 1991).

Described Species

Approximately 30 species of Angelonia are known,, but very few are mentioned in the

literature and even fewer have been cultivated. It is believed that A. angustifolia and A.

integerrima are the only cultivated species (Schoellhorn, 2002). Common names for this genus

include angelonia, summer snapdragon, and summer orchid (Hamrick, 2003). The following are

brief descriptions of species mentioned in the literature:

A. angustifolia Benth., a perennial species native to Mexico and the West Indies, grows

approximately 0.3 to 0.5 m tall and has glabrous stems, lanceolate leaves, and racemes

containing deep mauve to violet flowers. It flowers in the summer, has been naturalized in many

areas of the neotropics, and is the species most widely cultivated in commercial horticulture.

Two of the old, non-patented cultivars sold commercially are 'Alba', a white-flowered selection,

and 'Blue Pacific', a blue and white flowered selection (Bailey and Bailey, 1976; Griffiths, 1994;

Huxley, 1992; USDA, 2006).

A. cornigera Hook., an annual species native to Brazil, will grow about 0.3 m tall. Flowers

are located in the leaf axils as solitary flowers rather than as an inflorescence (Bailey, 1922).

A. gardneri Hook., a perennial species native to Brazil that will grow up to 1 m tall, has

glandular-pubescent stems and lanceolate-serrate leaves. It flowers in the summer and has

purple flowers with white centers (Griffiths, 1994; Huxley, 1992).

A. grandiflora Hort. was first introduced to gardeners in 1897 by Benary (Bailey, 1906). It

is believed to be a horticultural selection of A. salicariifolia.

A. integerrima Spreng., a perennial species native to southern Brazil and Paraguay, has

entire, lanceolate leaves and sky blue flowers containing purple spots. The upper corolla lip is









reduced and the lower corolla lip has ovate, coarsely-serrated lobes. It flowers in the summer

(Griffiths, 1994; Huxley, 1992).

A. procumbens, a native to eastern Brazil, has not been described in much detail except that

it shares similar characteristics with other species in this genus (Barringer, 1985).

A. salicariifolia Humb. & Bonpl., a perennial species native to Central America and the

West Indies, grows approximately 0.5 to 0.8 m (Bailey and Bailey, 1976). It has extremely

glandular, sticky-pubescent stems; broad, lanceolate, serrulate leaves; and mauve or bluish-

purple flowers. The only described cultivar is 'Grandiflora' (also known as A. grandiflora),

which has large white flowers that can reach 25 mm in size (Bailey and Bailey, 1976; Griffiths,

1994; Huxley, 1992; Pennell, 1920).

Cultivation

Angelonia is commonly grown outdoors as a perennial in tropical and subtropical regions

and as a summer annual in cooler climates (Huxley, 1992). It is commonly found growing along

damp edges of savannahs or in open, sunny locations in its native habitat. Multiple species are

located in the Caatinga, a tropical dry forest in northeastern Brazil, where rainfall is seasonal and

temperatures consistently range between 23 and 27 C (Machado et al., 1991).

The first known reference to its commercial availability is from Benary in 1897 (Bailey,

1906), but it was not widely popularized until the late 1990s. The high incidence of infection

with Cucumber Mosaic Virus limited its availability due to a lack of growers willing to grow

plants that could be virus infected (Hamrick, 2003). In the late 1990s, Ball FloraPlant released

multiple cultivars in the AngelMist series. These cuttings were touted as coming from 'clean'

stock plants that had undergone virus-indexing and subsequent virus eradication (Schoellhorn,

2002). Since then, multiple companies have released commercial series. A 'series' was a term

originally used to denote seed lines offered by one company that were very similar to each other









and varied only by one trait, such as flower color (Armitage, 1994). This term has since been

extended to describe a line of vegetatively-propagated cultivars available from a company.

Commercial Cultivars

Commercial cultivars are currently available from at least six different companies (Table

1-2). All of them are vegetatively-propagated except for the seed-propagated Serena series.

Through breeding advances, the range of flower colors available include white, pink, lavender,

dark blue, purple, and bi-colored. It is a versatile plant and can be used in mass landscape

plantings, hanging baskets, and mixed containers or as a cut flower. Variability is present in

angelonia for growth habit and flowering, including plant height, plant width, amount of leaf

pubescence, degree of leaf stickiness, number of days to first flower, flower color, flower size,

degree of reflexing in the two side lobes of the lower lip, inflorescence length, inflorescence

number, disease resistance, and length of flowering in landscape (Boldt, personal observation).

Angelonia is a warm season crop grown primarily for the spring and summer seasons in

southern states and for the summer season in northern states (Armitage, 1997). From a rooted

cutting, crop time will range from 6 to 10 weeks depending upon cultivar and greenhouse

conditions (Schoellhorn and Alvarez, 2002). Optimal growth occurs when plants are provided

with high irradiance levels, warm temperatures, and moderate fertilizer concentrations. Cultural

problems tend to be few as long as adequate greenhouse conditions are maintained. A more

detailed review of factors influencing angelonia production will be discussed in the following

sections.

Flowering

For most greenhouse crops, the onset of flowering is one of the most important steps in

determining that it is ready to be shipped to retail stores and garden centers. In nature, the timing

of flowering is important for plants so that they maximize their potential for successful









pollination and future survival (Bernier et al., 1993). In most species, environmental cues such

as photoperiod, temperature, and water availability are major factors that cue the transition from

a vegetative phase to a reproductive phase (Bernier et al., 1993). The major genetic pathways

controlling flowering in plants are the photoperiodic, vernalization, gibberellic acid (GA), and

autonomous pathways.

Plants are classified into three major groups with respect to their photoperiod response:

short-day plants flower when the night length is longer than a critical period, long-day plants

flower when the night length is shorter than a critical period, and day-neutral plants flower

regardless of the length of the night period (Roberts and Summerfield, 1987). Day neutral plants

are also termed 'photoperiod-insensitive' or 'autonomous-flowering' (Bernier et al., 1993;

Halevy, 1984).

The number of floricultural crops classified as day-neutral is small compared to the

number of photoperiodic-sensitive crops. Some of the most important crops are photoperiodic,

including Euphorbia pulcherrima (poinsettia), Dendranthema x grandiflorum (chrysanthemum),

and Kalanchoe blossfeldiana kalanchoee). In two recent studies, 68 percent of bedding plants

and 86 percent of Hibiscus species trialed were photoperiodic (Mattson and Erwin,2005; Warner

and Erwin, 2001 a).

A response to photoperiod, in general, is related to the latitude and elevation from which

the species originates. Day length changes consistently from year to year at a particular location

and can provide a consistent and accurate signal that the environment is favorable or unfavorable

for reproduction. The magnitude of the change in day length between seasons decreases the

closer one is to the equator (Vince-Prue, 1984). Thus, while photoperiod sensitive plants can be









found at all latitudes, it is common to find day-neutral plants near the equator (Wareing and

Galston, 1963).

In day-neutral plants, irradiance and temperature, as well as the interaction of the two, are

important factors that influence flowering (Bernier et al., 1993; Halevy, 1984). Light intensity

and quantity appear to be involved in floral initiation and portions of floral development, while

temperature is involved in the rate of floral development from initiation to anthesis (Adams,

1999; Kinet and Sachs, 1984).

Irradiance Influences Time to Flower

Light quantity refers to the total amount of light energy captured by a plant in a specified

time period and is often quantified as the daily light integral, or DLI (mol-m-2-d-1). Increasing the

light intensity will increase the DLI and amount of photosynthetically active radiation available

to the plant. Many day-neutral plants tend to have a slight quantitative long-day response since

increasing the daylength will increase the length of time per day that a plant can capture light

energy (Kinet and Sachs, 1984; Korczynski et al., 2002).

Across the United States, DLI varies from less than 5 molm-2-d-1 in northern states in

December to 55 to 60 molm-2-d-1 in the Southwest in July and August (Korczynski et al., 2002).

This wide range can greatly impact the time to flower for a crop and the length of time it

occupies greenhouse space. The influence of light intensity and light quantity has been observed

in a wide range of crops, and the general trend is that an increase in DLI will decrease the

number of days to anthesis. In irradiance-responsive crops, the impact of an increase in

irradiance will follow the law of diminishing returns. An identical increase in irradiance will

have a much greater impact on reducing time to flower on plants grown under a low DLI

compared to those grown under a higher DLI (Adams et al., 1997; Karlsson et al., 1989).









One way to quantify the effect of irradiance on flowering is to observe the number of days

to first flower in a species. Pelargonium x hortorum 'Radio' grown at 22 W-m-2 reached 100%

anthesis faster than those grown at 4 W-m-2 (Welander, 1983). Warner and Erwin (2005a)

observed a decrease in the number of days to first flower in Antirrhinum majus 'Rocket Rose',

Calendula officinalis 'Calypso Orange', Mimulus x hybridus 'Mystic Yellow', and Torenia

fournieri 'Clown Burgundy' as DLI increased. Pietsch et al. (1995) observed a decrease in

number of days to flower in Catharanthus roseus 'Grape Cooler' plants supplied with

supplemental lighting. Fausey et al. (2005) reported that an increase in DLI had a minimal

impact on the time to flower in three perennials: Achillea and Gaura plants grown as 22 C

flowered 5 and 7 d earlier, respectively, as DLI increased from 5 to 20 mol-m-2d-1 but no

difference was observed in Lavandula.

A second way to quantify time to flower is to report the rate of progress towards flowering,

defined as the reciprocal of the number of days to first flower (1/days to flower). Tagetes erecta

'Bonanza Yellow' had an increased rate of progress towards flowering as irradiance increased

from 5 to 25 mol-m-2d-1 (Moccaldi and Runkle, 2007), and chrysanthemum 'Resiliance' had a

higher rate of floral development under increasing photosynthetic photon flux (PPF) levels

(Warrington and Norton, 1991).

Temperature Influences Time to Flower

Floral development is a metabolically-driven, temperature-dependent process. From a base

temperature to an optimal temperature, any increase in temperature will increase the rate of

progress towards flowering. Above the optimal temperature, an increase in temperature will

decrease the rate of progress towards flowering (Armitage, 1994; Roberts and Summerfield,

1987). Karlsson and Werner (2001) observed that cyclamen grown at a constant temperature for

eight weeks following formation of visible buds flowered progressively earlier as temperatures









increased from 8 to 20 C, but later as temperatures increased from 20 to 24 C. The rate of

progress towards flowering increased linearly up to an optimum of 21.7 C in Viola x

wittrockiana 'Universal Violet' (Adams et al., 1997) and increased quadratically up to

approximately 25 C in Celosia 'Gloria Mix' (Pramuk and Runkle, 2005).

With many crops, the specific combination of day and night temperatures does not affect

time to flower, but rather average daily temperature (ADT) is a better indicator of the rate of

floral development. The optimal ADT temperature for flowering in chrysanthemum ranges

between 17 and 22 C (Van der Ploeg and Heuvelink, 2006). Campanula carpatica 'Blue Clips'

flowered approximately 20 d earlier as ADT increased from 15 to 25 C (Niu et al., 2001),

Catharanthus roseus 'Grape Cooler' flowered 30 d earlier between 18 and 35 C (Pietsch et al.,

1995), Platycodon grandiflorus 'Astra Blue' flowered 63 d earlier between 14 C and

29 C (Park et al., 1998), and Petunia xhybrida 'Wave Purple' and Viola x wittrockiana 'Sorbet

Blackberry Cream' flowered 67 and 23 d earlier between 12 and 24 C, respectively (Mattson

and Erwin, 2003). The rate of progress towards flowering in Salvia splendens 'Vista Red' and

Tagetes erecta 'Bonanza Yellow' increased under higher temperatures (Moccaldi and Runkle,

2007).

Temperature x Irradiance Interactions Influence Time to Flower

Temperature and light may interact to influence the time to flower. This interaction may

hasten or delay flowering, or the effects of each other may be negated. In general, an increase in

irradiance at lower temperatures is often more pronounced than at higher temperatures. In

Petunia xhybrida 'Snow Cloud' plants grown under either 6.5 or 13 mol-m-2-d-1, the higher DLI

resulted in earlier flowering at all temperatures except at the two highest temperature treatments

(25 and 30 C) (Kaczperski et al., 1991).









In Salvia splendens 'Vista Red', as temperature increased from 15 to 25 C, the number of

days to flower decreased from 42 to 24 d at 10 mol-m-2d-1 and from 37 to 21 d at 20 mol-m-2d-1

(Moccaldi and Runkle, 2007). In contrast, Impatiens wallerana 'Accent Red' plants grown with

less than 15 mol.m-2-d1 flowered earlier as temperature increased from 14 to 28 C, but no

difference between temperatures was observed at the highest DLI treatments (Pramuk and

Runkle, 2005). These differences may be attributed to the fact that salvia is a full-sun plant and

impatiens prefer partial-shade locations in the landscape, and at the highest light treatments, the

impatiens were exposed to supra-optimal DLI levels.

Flowering in Angelonia

Angelonia has been classified as a day-neutral crop. In a study by Miller and Armitage

(2002), five AngelMist cultivars were grown under natural days, continuous short-days (SD),

continuous long-days (LD), or various combinations of SD and LD (2 wk SD then LD, 4 wk SD

then LD, 6 wk SD then LD, 2 wk LD then SD, 4 wk LD then SD, or 6 wk LD then SD), and the

number of days to visible bud and anthesis were not affected by photoperiod treatment. It a

preliminary report, they noted that all cultivars consistently flowered between the 9th and 11th

nodes (Armitage et al., 2000). Starman (2001) reported that 'Blue Pacific' was also a day-

neutral cultivar. These results are consistent with the fact that the native range for angelonia is

tropical and subtropical regions. In the Caatinga (northeastern Brazil, 363'W 804'S), where a

number of field studies were conducted on flowering time and type of reproduction (Machado et

al., 2006; Vogel and Machado, 1991), the natural photoperiod range is from 11.5 h in mid-June

to 12.5 h in mid-December (U.S. Naval Observatory, 2008).

Time to flower is angelonia is influenced by both temperature and irradiance. Angelonia is

responsive to supplemental irradiance (Starman, 2001) and exhibits a quadratic response (Miller

and Armitage, 2002). 'AngelMist Pink' grown under natural days or natural days plus either 900









or 1200 gmol-m-2's-1 continuous supplemental lighting from high intensity discharge (HID) lights

flowered in 57, 49, and 45 d, respectively. In production, recommended light levels range

between 6,000 to 10,000 foot candles (approximately 1200 to 2000 gmol.m-2.S-1) (Ball

FloraPlant, 2006). An increase in temperature, up to an optimum, hastens flowering.

'AngelMist Pink' grown at constant temperatures of 15, 22 or 30 C flowered in 58, 47, and 50

d, respectively, and exhibited a quadratic response (Miller and Armitage, 2002). This would

indicate that the optimal temperature for flowering is between 22 and 30 C. It is unknown if

other cultivars have similar responses to temperature as 'AngelMist Pink.' In production, the

recommended growing temperatures are 17 to 20 OC night and 25 to 30 OC day (Ball FloraPlant,

2006). No studies have been published to date on the combined influence of temperature and

irradiance on time to flower in angelonia.

Factors Influencing Continued Flowering

In most warm-season crops, temperatures above 30 C will result in a decrease in flower

number (Armitage, 1994), but this threshold value will vary depending upon genus, species, and

even cultivar. Also, the length of the high temperature stress will influence the degree of

response observed.

This decline in flowering has been attributed to a carbon shortage within the plant. At

higher temperatures, especially high night temperatures, plants will have a higher rate of

photorespiration and a lower daily net photosynthesis. Flowers and developing inflorescences

will compete with other active sinks in the plant for a limited carbon supply, and over time,

fewer flowers will be able to develop. High temperature stress on flowering has also been shown

to influence signaling within the floral pathways. In Arabidopsis thaliana, floral abortion

occurred when whole plants or inflorescences only were subjected to temperatures above 33 C

(Warner and Erwin, 2005b). Based on a degree-hours (C-h) model, flower buds began aborting









between 200 and 300 C-h and complete inflorescences aborted above 300 C-h. Thus, while

less carbon assimilation at high temperatures influences flowering in many crops, there are most

likely other crops in which the inflorescence detects changes in temperature and translocates

signals within the plant leading to floral abortion.

Most of the published research on the influence of high temperatures on flowering has

been conducted on vegetable and field crops, in which a reduction in flowering or fruit set results

in decreased yield. In three field-grown summer brassica (canola) species (Brassica napus, B.

rapa, and B. juncea), an increase in temperature resulted in lower flower number, and

temperatures above 29.5 C resulted in significant yield loss (Morrison and Stewart, 2002). In

pepper (Capsicum annuum), a comparison of plants grown at 25 and 33 C indicated that the

number of flower buds and mature flowers was not affected by the increase in temperature, but

fruit set was less at 33 C (Erickson and Markhart, 2001). This would indicate that while flowers

developed, there was a subsequent problem with pollen development, ovary development, and/or

fertilization. In tomato (Lycopersicon esculentum), chronic high temperatures influence pollen

grain development and leads to reduced fruit set (Sato et al., 2000).

Studies looking at the impact of temperature on flower number or flower bud abortion in

greenhouse crops have been limited to time of first flower. This is of importance because

decreased flowering will impact plant quality and crop marketability. Kaczperski et al. (1991)

attributed a delay in flowering in Petunia xhybrida at temperatures above 25 C to floral bud

abortion. In a study conducted by Warner and Erwin (2005a) on five ornamental plant species,

the number of flower buds present at time of first flower was less for plants grown at 32 C

relative to 20 C. Impatiens wallerana was the least sensitive, with a 30 percent decrease in

flower bud number, and Toreniafournieri was the most sensitive, with a 95 percent decrease. In









a screen of 12 Viola x wittrockiana cultivars, the number of flower buds present at time of first

flower was 20 to 77 percent less in those plants grown at 30 rather than 20 C (Warner and

Erwin, 2006).

Angelonia cultivars were observed to stop flowering or have significantly fewer

inflorescences per plant in the University of Florida trial garden during the summer 2005 and

2006 seasons (Boldt, personal observation). This reduction in flowering has not been observed

in northern states, where it is planted in early summer and flowering is continuous until first

frost. It is unknown if the reduction in or cessation of flowering in angelonia cultivars is due to

chronic high temperature stress or advanced plant age since it is planted outside much earlier in

the year in the southern states relative to northern states. In a study by Miller et al. (2001), net

photosynthesis in two angelonia cultivars was optimal at 20.8 for 'AngelMist Purple Stripe' and

19.8 C for 'AngelMist Deep Plum'. In locations where both the day and night temperatures

exceed this range during the summer months, it could be expected that flowering may be reduced

due to lower net carbon assimilation.

Plant Growth

Temperature

Temperature has an impact on plant growth, flowering, and plant quality. It is known to

affect plant height, number of lateral shoots, flowering time, flower number, and flower size. In

many warm season crops, temperatures below 10 C will significantly slow plant growth and

temperatures above 30 C will cause a reduction in plant quality (Armitage, 1994).

Temperature impacts plant height by altering the number of nodes produced in a specified

time period and by altering internode length (Warner and Erwin, 200 b). As temperature

increases within the optimal range, the leaf unfolding rate will increase and number of nodes

produced by a plant will increase. This process is dependent upon average daily temperature, not









day or night temperatures, and it will reach a plateau at approximately 24.4 to 26.6 C (76 to 80

F). Intemode length in plants is affected not by average day and night temperatures, but rather

the magnitude of the difference between them. This is termed DIF, and the greater the DIF

(higher day relative to night temperature), the greater the intemode length (Erwin et al., 1989).

A study by Miller and Armitage (2002) reported that angelonia plant quality was greatest

at 22 C when plants were grown at a constant 15, 22, or 30 C. Plants grown at 15 oC were

chlorotic and stunted, while plants grown at 30 oC were tall with thin, brittle stems. Plant height

was 31, 52, and 68 cm, respectively, and responded linearly to temperature. This is similar to

results observed in Petunia xhybrida 'Snow Cloud' by Kaczperski et al. (1991), in which plant

height increased linearly and internode length increased quadratically as temperature increased.

In addition to plant height, temperature can affect flower size and flower number. A

decrease in flower size in response to increased temperatures has been observed in

Chn ytani/he'un morifolium (Willits and Bailey, 2000); Calendula officinalis, Impatiens

wallerana, Mimulus x hybridus, and Toreniafournieri (Warner and Erwin, 2005a); and Viola x

wittrockiana (Warner and Erwin, 2006). The number of flowers per plant has been shown to

decrease in Viola x wittrockiana and Petunia x hybrida, increase in Impatiens wallerana, and not

change in Viola x wittrockiana in response to increased temperatures (Mattson and Erwin, 2003;

Warner and Erwin, 2006).

Irradiance

Increased irradiance is generally associated with an increase in plant quality due to

improved plant architecture and an increase in flower size. Fausey et al. (2005) observed an

increase in lateral branching ofAchillea, Gaura, and Lavandula as DLI increased from 5 to 20

molm-2-d-1 and a two to three-fold increase in the number of inflorescences on Achillea and

Gaura. A linear increase in flower size was reported for cyclamen plants grown at 20 oC as DLI









increased from 1.8 to 21.6 mol-m-2-d- (Karlsson et al., 1989). An increase in flower diameter

has been reported for Calendula officinalis, Impatiens wallerana, Mimulus x hybridus, and

Toreniafournieri when grown at 21.8 versus 10.5 mol-m-2d-1 (Warner and Erwin, 2005).

Supplemental lighting or a natural increase in irradiance has been shown to increase

angelonia plant quality. 'Blue Pacific' plants grown under HID supplemental lighting were more

compact and better branched (Starman, 2001). Holcombe et al. (2001) have reported that

angelonia plants grown under 6 mol-m-2d-1 had very weak lateral branches, plants grown at 12

mol-m-2d-1 were of acceptable quality, but 18 mol-m-2d-1 was necessary for an adequate number

of inflorescences.

Temperature x Irradiance Interactions

The interaction of temperature and light can have a major impact on plant quality. In

Gypsophilapaniculata L., the temperature corresponding to the highest plant quality decreased

under higher irradiance levels. At a light level of 450 tmol-m-2'-1, the temperature optimum

was 20 C, but at 710 tmol-m-2'-1, the temperature optimum was between 12 and 20 C

(Hinkleton et al., 1993). In Salvia splendens 'Vista Red', however, the temperature

corresponding to maximum plant height increased as irradiance increased. At 5 mol-m-2-d1,

plant height was greatest at 20 OC, while at 25 mol-m-2-d1, plant height was greatest at 24 C

(Moccaldi and Pramuk, 2007). It is unknown whether or not this increase in plant height was a

desirable or undesirable characteristic for this crop.

In angelonia, no research has been published on whether temperature and light have an

interactive effect on plant height or overall plant quality. Angelonia plants grown under natural

days in the spring and summer months will be exposed to increasing temperature and light levels,

but by varying amounts.









Fertilizers


Plant Growth

Use of the correct fertilizer and concentration during crop production can significantly

impact final quality of the finished product. Plant height, plant size or fullness, leaf color,

number of flowers or inflorescences, time to flower, and dry weight can be impacted by the type

and concentration of fertilizer applied. Growth parameters, such as height and dry weight, tend

to increase in response to increasing fertilization up to an optimum, and then decrease (James

and van lersal, 2001). Flowering has been shown to be delayed at low nutrient concentrations in

Salvia splendens (Kang and van lersal, 2004).

The optimal fertilizer concentration to use in production varies by crop, cultivar, irrigation

method, frequency of application, current nutritional status of the crop, and environmental

conditions during crop production. It has been shown that the light level during production does

not influence the optimal fertilizer concentration for Begonia semperflorens and Petunia

xhybrida, examples of low-fertility and high-fertility requiring crops (Nemali and van lersal,

2004), but temperature influences optimal fertilization in petunia (Kang and van lersal, 2001).

As temperature increased, the optimal concentration necessary for plant growth decreased.

Fertilizer recommendations vary depending upon irrigation method. Leaching does not

occur in a subirrigation system and the soluble salts applied are either used by the plant or

accumulated in the potting medium. However, leaching can occur with overhead irrigation and

excess salts may be leached as needed to maintain the correct electrical conductivity (EC) range

for optimal plant growth. It has been recommended that subirrigated crops are fertilized with a

half-strength solution relative to the suggested concentration for overhead watering (Nelson,

1994). Klock-Moore and Broschat (2001) observed that subirrigated petunias at fertilizer









concentrations of 50, 100, or 150 mg-L-1 had final EC levels that were twice as high as overhead

irrigated plants at 50 and 100 mg-L-1 and five times as high at 150 mg-L1.

Crops vary in their sensitivity to fertilization. Optimal growth has been observed to

range from N at approximately 100 mg-L-1 for Celosia argentea, Impatiens xhawkeri, Impatiens

wallerana, and Zinnia elegans (Kang and van lersal, 2002; Kent and Reed, 1996; Whipker et al.,

1999) to up to 400 mg-L-1 in Gomphrena globosa and Matthiola incana (Kang and van lersal,

2002). Different plant quality parameters may have different optimal concentrations and a

grower will need to determine which factor is of most importance. For instance, Petunia

xhybrida dry weight was maximized when supplied with a complete fertilizer with N at 355

mg-L-1, height at 255 mg-L-1, and flower number at 165 mg-L-1 (James and van lersal, 2001).

Within a crop, cultivars may have different fertilizer optimums. Double impatiens (Impatiens

wallerana) 'Purple Magic', a green-leaved cultivar, required a higher concentration than

'Blackberry Ice', a variegated-leaved cultivar (Whipker et al., 1999). Cultivar differences have

been noted in poinsettia (Euphorbiapulcherimma) related with leaf color. Dark-green leaved

cultivars require an electrical conductivity (EC) of 1.5 to 2.0 mmhos-cm-1 (mS-cm-1) using the

saturated media extract (SME) procedure, whereas medium-green leaved cultivars require 2.0 to

2.5 mmhos-cm-1 (Ecke et al., 2004).

Use of a Hand-held Meter to Measure Chlorophyll Content

The SPAD meter (Minolta Co., Osaka, Japan) is an instrument used to measure leaf

chlorophyll content of a crop quickly and non-destructively. Output values do not have a unit

associated with them, but they can be correlated with leaf chlorophyll content, with increasing

values indicating higher chlorophyll levels. The nature of the relationship varies. It has been

reported to be linear for ornamental foliage plants (Wang et al., 2005); non-linear for Oryza

sativa, Glycine max, Triticum aestivum, and Solanum tuberosum (Monje and Bugbee, 1992;









Uddling et al., 2007); and exponential for Glycine max and Zea mays (Markwell et al., 1995).

SPAD values have been shown to correlate strongly with the actual chlorophyll content of leaves

as determined through destructive sampling (r2 values of approximately 0.9 or greater) (Monje

and Bugbee, 1992; Uddling et al., 2007; Wang et al., 2005).

SPAD meters were initially used in field crops to determine if supplemental N was

necessary for optimal yield. Nitrogen deficiencies show up as lighter green foliage coloration

due to a decrease in the chlorophyll content of the leaf. Greenhouse and bedding crops grown in

soilless media have their fertilizer requirements met through the application of a water-soluble or

slow-release fertilizer. These tend to be complete fertilizers, but their concentrations are

reported as N concentration in mg-L-1 or ppm. Increasing the fertilizer N concentration applied

will result in an identical increase in concentration of all other elements present and an increase

in the total concentration of soluble salts available to the crop. In angelonia, when single

elements were withheld from an otherwise complete fertilizer, deficiency symptoms for N, P,

and K all appeared at the same number of days after start of treatment (Williams, 2004). Thus,

the use of SPAD meter can indicate that plants with lower values will likely have lower tissue

concentrations of other elements as well.

Cultivar and environmental factors can influence SPAD readings, including normal leaf

coloration under adequate N fertilization, relative water status of the plant, temperature, and

sunlight (Peterson et al., 1993). Leaf size should not result in significant variation between

samples since the measurement area for the meter is very small, 2 mm by 3 mm (Minolta, 1989).

Leaf thickness ranging from 0.19 to 0.66 mm did not affect SPAD readings (Wang et al., 2005)

and Minolta states that the meter can determine a value for leaves up to 1.2 mm thick (Minolta,

1989). Markwell et al. (1995) reported that the exponential equation developed for soybeans and









corn fit data collected for sorghum and Arabidopsis and suggested that multiple crops may have

very similar correlations between the SPAD value and actual chlorophyll content. Wang et al.

(2005), however, reported that linear models developed for 10 different foliage crops were not

identical and that individual models should be developed for use across crops, but may be useful

for comparing cultivars.

Published fertilizer guidelines for angelonia vary by company. All recommend constant

liquid feeding with a complete fertilizer, but the level of fertilization ranges from N at 150 to 250

mg-L-1 (Ball FloraPlant, 2006; Fischer, 2008; PanAmerican Seed, 2006; Proven Winners, 2006).

Fertilizer guidelines published in trade press articles recommend N at concentrations ranging

from 75 to 200 mg-L-1 (Armitage, 1997; Schoellhorn and Alvarez, 2002; Smith, 2007). It is

unknown whether these differences in recommended levels are due to cultivar, series, or

environmental conditions.

Plant Growth Regulators

Introduction

Plant growth regulators (PGRs) are used in the floriculture industry to control plant growth

during production, improve quality, and increase longevity at retail (Arteca, 1996). These

chemicals inhibit stem elongation without killing the meristem (Cathey, 1964), and plants treated

with a PGR look identical to untreated plants except for the fact that they are shorter. The first

PGRs were introduced in the 1960s, and since then two major groups have arisen based on mode

of action. One group inhibits stem elongation by inhibiting various steps in the gibberellin

synthesis pathway and this includes daminozide and paclobutrazol. The second group releases

ethylene within the plant, resulting in an inhibition of stem elongation and an increase in lateral

branching in certain crops, and this group includes the chemical ethephon (Rademacher, 2000).









The effect that a PGR will have on a plant is dependent upon multiple factors, including

species sensitivity, cultivar sensitivity, plant age, PGR concentration, method of application, and

environmental conditions under which the plant is grown (Barrett, 2001; Fletcher et al., 2000;

Rademacher, 2000). Crops may exhibit sensitivity to some, all, or none of the PGRs typically

used in production. For example, paclobutrazol inhibited stem elongation in Hibiscus coccineus,

H radiatus, and H trionum but daminozide was effective only on H trionum (Warner and

Erwin, 2003). In a trial of 26 ornamental cabbage and kale cultivars conducted by Gibson and

Whipker (2001a), only two cultivars were responsive at 2500 mg-L-1 and eight at 5000 mg-L1.

Plant age can impact responsiveness to a PGR application, especially for foliar-applied

chemicals, since more chemical is taken up by younger leaves due to the presence of a thinner

cuticle (Sachs and Hackett, 1972).

The environmental conditions in which the crop is grown can greatly affect efficacy. In

warmer temperatures, plants are growing at a faster rate than those grown at cooler temperatures.

They usually will require the application of a higher concentration or multiple applications in

order to see a similar amount of height control relative to those grown at cooler temperatures

(Barrett, 2001). Growers typically adjust the concentration of a PGR application between winter

and summer applications.

The following sections will describe in greater detail each of the three PGRs to be used in

this research: paclobutrazol, daminozide, and ethephon. Reported similarities and differences

among crops and between cultivars will be discussed, as well as any known results pertaining to

angelonia cultivars.

Paclobutrazol

Paclobutrazol [(2RS,3RS)-1 -(4-chlorophenyl)-4,4-dimethyl-2-(1H-1,2,4-triazol- 1-

yl)pentan-3-ol], discovered in the 1960s during trialing for new fungicides (Fletcher et al., 2000),









is a member of the triazole family. Trade names for paclobutrazol include Bonzi, Downsize,

Paczol, Piccolo, and PPP-333. It inhibits gibberellin (GA) synthesis by inhibiting the function of

cytochrome P450 and affecting the formation of ent-kaurenoic acid from ent-kaurene, GA

precursors (Fletcher, 2000; Rademacher, 2000). Treated plants will exhibit shorter internodes

and thicker, darker green leaves. In chrysanthemum, this increased leaf thickness has been

reported to be due to the formation of an additional layer of palisade cells (Burrows et al., 1992).

Paclobutrazol, effective as a media drench or foliar spray, is translocated through the

xylem only (Barrett, 2001). The application of a foliar spray will be effective at inhibiting

internode elongation only if it comes in contact with stem tissue and developing meristems, so

good spray coverage is essential (Barrett, 2001). Media drenches are highly effective and very

low concentrations are needed compared to foliar sprays. Paclobutrazol is not very water-soluble

and will rapidly move into the wax layers present in the leaf. In a study by Chamel et al. (1991),

paclobutrazol uptake by leaves was determined but no distinction was made as to whether it was

adsorbed and/or absorbed, thus they referred it as 'soprtion'. Sorption was rapid during the first

hour following application and almost all was sorbed within 24 h of application. These results

concur with findings from Barrett et al. (1987) in which the efficacy of a spray application was

not reduced when plants were overhead irrigated as soon as 30 minutes after application. While

paclobutrazol rapidly moves out of solution to the cuticle, Chamel et al. (1991) determined that

only a very small amount, approximately 5 percent, penetrates the cuticle and into the leaf.

The timing of an application will have an impact on the amount of efficacy observed.

When Chlyii/wi'uniii 'Bright Golden Anne' plants were sprayed at 0, 2, or 4 weeks following

pinch, the earlier applications resulted in increased efficacy and shorter plants at flowering

relative to the later applications and untreated plants (Gilbertz, 1992). This increased efficacy









early in the crop may be due to a combination of factors, but most likely is due to the reduced

canopy size at the time of application. When applied at pinch, it is easier to get good spray

contact with the stem and more soil media will be sprayed, resulting in a slight drench effect.

Recommended concentrations for crops range between 5 and 90 mg-L-1 for foliar sprays

and 1 mg-L-1 for media drenches (Syngenta, 2005) and may be adjusted depending upon species

or cultivar sensitivity. The triazoles are very active and usually only very low concentrations are

necessary for efficacy (Barrett, 2001). Paclobutrazol is less active than uniconazole [(E)(S)-1(4-

chlorophenyl)4,4-dimethyl-2(1,2,4-triazol- -yl)pent- 1-ene-3-01] and two to four times as much

is needed for similar efficacy, but it is much more active than daminozide. In Scaevola aemula,

a media drench of 1 mg-L-1 uniconazole had similar efficacy as a 4 mg-L-1 paclobutrazol drench

(Starman and Williams, 2000). Paclobutrazol and uniconazole sprays at 30 and 15

mg-L-1, respectively, provided similar height control in Ch/y ,lwmii/,u,n, 'Bright Golden Anne'

(Gilbertz, 1992).

Cultivar sensitivity has been reported in ornamental cabbage and kale, with 'Osaka

White' exhibiting a linear response and 'Nagoya Red' exhibiting a linear-plateau response to

increasing concentrations of paclobutrazol (Gibson and Whipker, 200 b). This indicates that

'Osaka White' is less sensitive than 'Nagoya Red.' Potential differences in cultivar sensitivity

have been reported in angelonia. A spray concentration of 50 mg-L-1 was effective for

'AngelMist Pink' (Miller and Armitage, 2002), but concentrations up to 80 mg-L-1 were

ineffective for 'Blue Pacific' (Starman, 2001). However, it is not known whether these

differences can be explained by cultivar sensitivity or differences in environmental conditions

during production. No recommendations for foliar sprays have been published in cultural

guidelines or the trade press, but drench recommendations vary from 1 to 16 mg-L-1 (Armitage,









1997; PanAmerican Seed, 2006; Schoellhorn and Alvarez, 2002). Proven Winners (2006)

recommends the application of 2 to 5 mg-L1 uniconazole on 'Angelface Blue' and 5 to 10 mg-L-1

on 'Angelface White' and 'Angelface Blue Bicolor', indicating that differences in cultivar

sensitivity likely exist in this series. However, it is not known if other cultivars differ within

series and/or across series.

Daminozide

Daminozide [butanedioic acid mono (2,2-dimethylhydrazide)] was first described in 1962

and is known by the trade name B-Nine (Plant Growth Regulator Working Group, 1977).

Treated plants exhibit shorter internodes and are smaller than untreated plants due to its effect on

inhibiting the synthesis of GA (Rademacher, 2000). It is much less active than the triazoles and

in responsive species, concentrations of 2500 to 5000 mg-L-1 are recommended (Chemtura,

2001). Daminozide is applied as a spray and is very mobile within the plant (Barrett, 2001).

Unlike paclobutrazol, it is very water soluble and moves slowly from the aqueous solution to the

cuticle. It can be washed off if leaves are watered within 4 h following application (Barrett et al.,

1987). Due to the high concentration needed for efficacy, daminozide is more effective when

used in winter months or combined in a tank mix with chlormequat chloride [chlormequat (2-

chloroethyl)trimethylammonium chloride; Cycocel] (Barrett 2001; Cathey, 1975).

Plant response to daminozide varies with crop and cultivar. In a study by Gibson and

Whipker (2000a) on 26 ornamental cabbage and kale cultivars, two were responsive at 2500 and

eight at 5000 mg-L-1. Lewis et al. (2004) observed a cultivar differences in pansy in which

'Colossus Yellow Blotch' and 'Delta Pure Yellow' were responsive at 4500 mg-L-1 but 'Majestic

Giants Purple' was unresponsive at all concentrations. In contrast, osteospermum

(Osteospermum ecklonis) cultivars 'Congo' and 'Wildside' both were insensitive at

concentrations of up to 10,000 mg-L-1 (Gibson and Whipker, 2003).









Angelonia 'AngelMist Pink' has shown a response to daminozide at 2500 and 5000

mg-L-1 and 'Blue Pacific' at 5000 mg-L-1 (Miller and Armitage, 2002; Starman, 2001). Selecta

First Class (2008) recommends daminozide at 2500 mg-L-1, and PanAmerican Seed (2006) and

Ball FloraPlant (2006) recommend tank mixes of daminozide and chlormequat chloride for

height control.

Ethephon

Ethephon (2-chlorophenylphosphonic acid) was first synthesized in 1946, but its

mechanism of decomposition was not reported until 1963 (Beaudry and Kays, 1988). Under

high pH conditions, it naturally breaks down to form chlorine, phosphonic acid, and ethylene

(Beaudry and Kays, 1988). The release of ethylene is the key by-product of this reaction which

affects plant growth. Ethylene, a simple hydrocarbon, is the smallest known plant hormone in

the plant and is involved in a myriad of plant processes throughout its life cycle, from seed

germination to plant senescence (Arteca, 1996). Ethephon is considered a plant growth regulator

because the release of ethylene in the plant can result in an inhibition of stem elongation,

increased lateral branching, and/or delayed flowering. It is marketed under the trade names of

Florel, Pistill, and Ethrel and applied at concentrations ranging from 250 to 1000 mg-L-1 (Barrett,

2001).

Ethylene is known to inhibit cell elongation and interact with auxins to affect the

outgrowth of lateral buds. Cells treated with ethylene will have a similar total volume relative to

untreated cells but their elongation is inhibited (Osborne, 1974). Burg (1973) reported that this

is due to a reorientation of the microfibrils in the cell wall such that longitudinal growth but not

lateral growth is restricted, explaining why ethylene-treated plants may have thicker stems. The

mechanism underlying the release of lateral buds from apical dominance and their subsequent

growth is believed to be due to a shift in hormone ratios within the plant. In Petunia xhybrida,









the application of ethephon to plants resulted in a 20 percent decrease in IAA (indole-3-acetic

acid) and a 24 percent decrease in the auxin:cytokinin ratio relative to untreated plants; no

changes were observed in cytokinins between treated and untreated plants (Haver and Schuch,

2001).

The efficacy of an ethephon application depends upon the plant and the conditions at the

time of application, including solution pH, temperature, relative humidity, plant species and

cultivar. Ethephon is stable below pH 5.0 but will begin to decompose and release ethylene in

higher pH environments (Warner and Leopold, 1969), with 98 percent decomposition seen in an

ethephon solution maintained at pH 7.4 at 25 C (Segall et al., 1991). In a study examining the

rate of decomposition of ethephon, it increased significantly between pH 6.0 and 8.0 at 25 C.

However, temperature greatly influenced the magnitude of the decomposition, with similar rates

observed between a solution with pH 9.1 kept at 30 oC and a solution with a pH of 6.1 kept at 40

C (Biddle et al., 1976). Differences between species and cultivars may be due to differences in

ethylene sensitivity or due to differences in the leaf morphology (Beaudry and Kays, 1988). In

leaves with a thinner cuticle, penetration of the chemical is greater than those with a thicker

cuticle.

Ethephon or ethylene can be translocated through the plant. Puech and Crane (1975)

observed the presence of 14C in the translocation stream within 2 h of an ethephon application,

but it was not reported if it was ethephon or the released ethylene that was being translocated.

Kwong and Lagerstedt (1977) observed the accumulation of 14C in apical meristems and nodes

1 h following application, but again, no distinction was made as to whether it was ethylene or

ethephon. In chrysanthemum, the application of a single drop of ethephon to a leaf was

significant enough to affect stem elongation and delay flowering, and this effect was still









observed if the treated leaves were removed 12 h following application, indicating that

translocation or signaling occurred within that time interval (Stanley and Cockshull, 1989).

Common responses to ethephon applications include a reduction in stem elongation, an

increase in lateral branching, and delayed flowering. Decreased plant height has been observed

in Achillea, Echinacea, Leucanthemum, Monarda, and Phytostegia (Hayashi et al., 2001). Foley

and Keever (1992) and Carpenter and Carlson (1972) observed an increase in the number of

laterals in geranium 'Hollywood Star' and 'Dark Red Irene' plants treated with 500 and 1000

mg-L-1, respectively. Glady et al. (2007) reported increased branching of Coreopsis verticillata

'Moonbeam', Dianthus caryophyllus 'Cinnamon Red Hots', and Veronica longifolia 'Sunny

Blue Border' following ethephon applications. Delayed flowering of more than 2 weeks has

been reported for ethephon-treated plants. Scaevola treated with ethephon at 500 and 1000

mg-L-1 flowered 8 and 11 d later than untreated plants, respectively (Starman and Williams,

2000), and geranium 'Dark Red Irene' treated with 1000 mg-L-1 flowered 16 d later (Carpenter

and Carlson, 1972).

One drawback to the use of ethephon is the appearance of phytotoxicity symptoms

following application in some, but not all crops. Hayashi et al. (2001) trialed eight species of

perennials and only Monarda exhibited leaf necrosis. Glady et al. (2007) observed deformed

growth and leaf necrosis in Dianthus caryophyllus 'Cinnamon Red Hots', but not in the two

other crops also in the experiment.

Cultivar differences have been reported following ethephon sprays. Hammond et al.

(2007) observed more compact growth and delayed flowering in Gaillardiapulchella 'Torch

Flame' but no difference in plant growth between treated and untreated plants of a native Florida

ecotype. In a study of 23 cultivars from 12 different genera, Faust and Lewis (2005) indicated









that crops differed in response, but all cultivars within a particular genus responded similarly. In

contrast, Starman et al. (2004), trialed 27 cultivars across 16 genera and observed cultivar

interactions for plant height, plant width, and number of days to flower. For example, treated

plants were significantly shorter than untreated plants for all cultivars ofDiascia xhybrida,

Impatiens wallerana, Lantana camera, and Nemesia xhybrida. In Antirrhinum majus,

Calibrachoa hybrid, and Petunia xhybrida, certain cultivars were responsive and others were

nonresponsive.

Ethephon has been reported to be an effective PGR for angelonia 'Blue Pacific' and no

phytotoxicity symptoms were reported (Starman, 2001). Proven Winners (2006) and Ball

FloraPlant (2006), however, recommend against the use of ethephon due to its potential to cause

leaf tip bum and distorted growth, but this has not been mentioned in the production guidelines

of other companies.

Objectives

1. Conduct a cultivar screen of commercial Angelonia angustifolia cultivars and determine the
range of variability present within this species for growth and flowering traits. Of interest
will be how variable the cultivars are within each series and across series. From this set of
cultivars, a sub-sample of cultivars representing this range of variability will be selected for
use in subsequent experiments.

2. Determine the effect of season on time to flower and plant growth on select cultivars grown
under natural days in the greenhouse during the main production window (late winter
through early summer).

3. Determine the effect of three plant dates on the summer landscape performance of select
cultivars.

4. Determine the effect of increasing fertilizer concentrations on time to flower, plant growth,
and flowering characteristics of select cultivars grown in the greenhouse.

5. Determine the effect of paclobutrazol, daminozide, and ethephon on growth of select
cultivars during greenhouse production and identify if they differ in their response to
chemical and/or concentration applied










Table 1-1. Current taxonomic classification ofAngelonia angustifolia.
Kingdom Planta
Division Magnoliophyta (Angiosperms)
Class Magnoliopsida (Dicots)
Subclass Asteridae
Order Scrophulariales
Family Plantaginaceae
Genus Angelonia Humb. & Bonpl.


Commercially available angelonia series.
Company
Danziger
Proven Winners
Selecta First Class
Ball Horticultural Company
Syngenta Flowers
Ball Horticultural Company


Propagation method
Vegetative cuttings
Vegetative cuttings
Vegetative cuttings
Vegetative cuttings
Vegetative cuttings
Seed


Table 1-2.
Series
Alonia
Angelface
Angelina
AngelMist
Carita
Serena









Table 1-3. Cultivar names of angelonia used in experiments.


Cultivar
Angelface Blue
Angelface Blue Bicolor
Angelface Dresden Blue
Angelface Pink
Angelface Wedgewood
Blue
Angelface White

Angelina Blue
Angelina Dark Blue
Angelina Pink
Angelina Pink and White
Angelina Violet and White
Angelina White

AngelMist Basket Pink

AngelMist Basket Purple

AngelMist Basket White

AngelMist Dark Lavender
AngelMist Dark Pink
AngelMist Dark Rose
AngelMist Deep Plum Imp.
AngelMist Lavender
AngelMist Lavender Stripe
AngelMist Pink
AngelMist Plum
AngelMist Purple
Improved
AngelMist Purple Stripe
AngelMist White Cloud
AngelMist White Improved


Status
Commercially available


Commercially
Commercially
Commercially

Discontinued

Commercially
Commercially
Commercially


available
available
available


Plant code
ANBLAUZWEI

ANSKY
ANPINK
ANWEDG

ANWHIT


Plant patent
PP 14,189

PP 17,155
PP 16,818
PPAF

PP 13,179


available
available
available


Commercially available
Commercially available

AngelMist Spreading
Pink
AngelMist Spreading
Purple
AngelMist Spreading
White
AngelMist Purple


Commercially
Commercially
Commercially
Commercially
Commercially
Commercially
Commerciallynued
Discontinued


available
available

available
available
available
available


Commercially available
Commercially available
Commercially available


BALANGBAKIN

BALANGBEKE

BALANGSPRI

BALANGDARLA
BALANGDARPI
BALANGDAROS

BALANGLADER
BALANGLAST
BALANGPIKIM
BALANGPLUM
BALANGIMPU

BALANGPRIPE

BALANGWITIM


PPAF

PP 15,546

PPAF


17,516
17,071
17,243

17,909
16,668
17,515
17,232
13,921


PP 16,501


Serena Lavender
Serena Lavender Pink
Serena Purple
Serena White
z Commercially available,
Y Seed propagated cultivar


Commercially
Commercially
Commercially
Commercially


discontinued, or renamed.


available
available
available
available


Noney
Noney
NoneY
Noney









CHAPTER 2
Angelonia angustifolia CULTIVAR SCREEN

Introduction

Angelonia, a genus native to Central and South America (Huxley, 1992), contains

approximately 30 species and Angelonia angustifolia is the only species grown commercially.

initially it was commercially available as non-patented cultivars, but its popularity was limited

due to the incidence of viruses. In the late 1990s, the first virus-indexed cultivars were

introduced and this crop saw a resurgence in popularity. Since then, multiple companies have

developed angelonia breeding programs and have released cultivars with a range of growth and

flowering characteristics and landscape performance.

The objective of the first experiment was to conduct a cultivar screen on 31 commercially

available cultivars from three companies and quantify the range of phenotypic variability present

in growth habit, time to first flower, flower size, and powdery mildew susceptibility. The second

experiment was designed to look at differences in flowering and growth habit in a small set of

cultivars over multiple production seasons. These cultivars were selected based upon an

observational cultivar screen conducted in summer 2006 and were selected for their range of

flowering and growth habits.

Materials and Methods

Experiment 2-1 Cultivar Screen of 31 Commercial Cultivars

Rooted cuttings or plugs (seedlings) of 31 commercially available Angelonia angustifolia

cultivars were received the second and third week of January 2007 (Ball FloraPlant, West

Chicago, IL; Costa Farms, Miami, FL; EuroAmerican Propagators, Bonsall, CA; Sunshine

Greenhouses, Provo, UT). Nine plants of each cultivar were planted on 25 Jan. into









2.2 L (16 cm diameter) pots filled with Fafard 2 soilless medium (Conrad Fafard, Inc., Apopka,

FL) and pinched to three nodes on 30 Jan. Plants were watered as needed and continuously

liquid fed with N at 150 mg-L-1 using a 20.0N-4.4P-16.6K fertilizer (Peters 20-10-20 Florida

Special, The Scotts Co., Marysville, OH). The experimental design was a complete randomized

block design, with three blocks and three plants of each cultivar per block. Over the length of

the experiment the average daily minimum, maximum, and 24-h temperatures were 17.0, 28.0,

and 21.4 C, respectively. The average daily light level was 20.2 mol-m-2-day-.

Plants were observed daily and the time from planting to first open flower and

marketability were recorded. An open flower was defined as having the corolla (both upper and

lower lips) unfurled. Marketability was defined as when a plant had at least 20 open flowers,

regardless of the number of inflorescences with open flowers. When each plant reached

marketability, plant height and width were measured and the number of inflorescences with open

flowers was determined. Plant height was measured from the top of the pot to the tallest growing

tip. Plant width was measured at the widest point in the plant canopy and then again

perpendicular to the first measurement. Average plant width was used for analysis. To provide

an additional comparison of the characteristics for the cultivars at the same point in time, on 6

Apr. (9.5 weeks after pinch), the number of inflorescences with at least one open flower was

counted, flower height and width was measured with calipers, and plants were evaluated for

powdery mildew susceptibility. Powdery mildew susceptibility was rated on a 1-5 scale with 1 =

no powdery mildew present, 2 = slight presence of powdery mildew, 3 = moderate presence of

powdery mildew, 4 = severe presence of powdery mildew, and 5 = leaves completely covered

with powdery mildew.









Data were analyzed using SAS 9.1 (SAS Institute, Cary, NC) for analysis of variance and

mean separation using Waller-Duncan at a=0.05.

Experiment 2-2 Cultivar Comparison of Seven Cultivars across Multiple Seasons

Seven Angelonia angustifolia cultivars were selected from a preliminary screen of 25

cultivars conducted in July and August 2006 in order to encompass a wide range of growth

habits, flower size, and time to flower (Table 2-1). These cultivars are the following: 'Angelina

Violet and White', 'AngelMist Dark Lavender', 'AngelMist Lavender Stripe', 'AngelMist

Purple Improved', 'AngelMist Purple Stripe', 'Angelface Blue', and 'Angelface White'. Each

cultivar was propagated from unrooted cuttings and grown to finished size at four different

seasons: late winter, early spring, late spring, and early summer.

Plants for the late winter experiment were the same as those used in Expt. 2-1. For the

other three seasons, stock plants were grown in 2.2 L (16 cm diameter) pots at the University of

Florida (Gainesville, FL) on a subirrigation bench and continuously liquid fed with a 20.ON-

4.4P-16.6K fertilizer (Peter's 20-10-20 Florida Special, The Scotts Co., Marysville, OH) with N

at 75 mg-L-1. Stock plants were replaced every three months with new virus-indexed plants

obtained from the breeder companies in order to maintain clean stock. Cuttings of each cultivar

were harvested and stuck in 25 mm moistened Ellepots (Knox Nursery, Winter Garden, Florida)

on the following dates: 19 Feb., 28 Mar., and 9 May 2007. They were placed under intermittent

mist for 2 to 3 weeks and acclimatized in a natural ventilated greenhouse for 1 week. During

acclimatization, rooted liners were watered as needed and continuously liquid fed with N at 150

mg-L-1 using 20.0N-4.4P-16.6K. Rooted liners (one plant per pot) were planted on 25 Jan., 14

Mar., 19 Apr., and 31 May into 2.2 L pots filled with Fafard 2 soilless medium (Conrad Fafard,

Inc., Apopka, FL) and pinched to three nodes 5 to 7 days later. Day 0 for each of the four

seasons was 30 Jan., 21 Mar., 25 Apr., and 6 June. Plants were watered as needed and









continuously liquid fed with N at 150 mg-L-1 using a 20.0N-4.4P-16.6K fertilizer (Peters 20-10-

20 Florida Special, The Scotts Co., Marysville, OH). Plants were observed daily and the date for

first open flower and marketability were recorded. At marketability, plant height was measured.

Once all plants had reached first flower, flower height and width was measured using calipers,

and plants were evaluated for presence of powdery mildew (see exp. 1 for explanation of data

collection). Temperature and light levels were recorded continuously over the length of the

experiment using a HOBO Microstation Data Logger (Onset Computer Corp., Bourne, MA), and

average daily minimum, maximum, and 24-hr temperatures and daily light integral were

determined for each season (Table 2-2).

The experimental design was a split plot, with season as the main plot and cultivar as the

sub-plot. Within each season, all cultivars were grown on a single bench in a complete random

block design with three blocks. The late winter season had 9 plants and the other three seasons

had 12 plants as the experimental unit. Data were analyzed using PROC MIXED in SAS 9.1

(SAS Institute, Cary, NC) and mean separation was performed using Tukey's at a=0.05.

Results and Discussion

Experiment 2-1

Days to first flower

Angelonia angustifolia cultivars flowered, on average, 52.5 days after pinch (DAP). The

earliest-flowering cultivar was 'AngelMist Dark Lavender' at 41.4 d and the latest was

'Angelface Blue Bicolor' at 63.8 d (Table 2-3), a difference of almost 23 d.

Cultivars within the Serena series were the most uniform and the range between the earliest

and latest-flowering cultivar was approximately 1 d. The range between the first and last cultivar

to flower in the Angelface and Angelina series were 10 d each. The AngelMist series had the

widest range of flowering times, 18 d from the first to the last cultivar to flower. All Angelface









and Serena cultivars took longer to flower than the average, and Angelina cultivars flowered

more quickly than the average.

The uniformity observed in the Serena series is consistent with the fact that it is seed

propagated and the other three series are vegetatively propagated. Plants of a vegetatively-

propagated cultivar are very uniform, but cultivars within a vegetatively-propagated series

usually have more genetic variability than seed propagated cultivars within a series (personal

observation). These differences can have a great effect on crop production. Miller and Armitage

(2002) reported that five cultivars within the AngelMist series responded similarly to

temperature, irradiance, photoperiod, and growth retardants. However, Expt. 2-1 contained 15

cultivars from that series and they all did not flower uniformly. The wide variability seen

between cultivars in the AngelMist series relative to the other two vegetatively-propagated series

may be due to the fact that 15 cultivars were trialed in this experiment while the others had six

cultivars each.

Days to marketability

Cultivars reached marketability between 45.4 and 69.8 DAP, a range of more than 24 d

(Table 2-3) and averaged 58.3 d. 'AngelMist Dark Lavender' and 'Angelface Blue Bicolor'

were the earliest and latest to marketability, respectively. This is the same as observed for time

to flower. The degree of uniformity within each series followed the same trends observed with

time to first flower.

All cultivars progressed from first flower to marketability in 4 to 8 d. Serena cultivars

flowered the quickest after reaching first flower, and 'Angelina Blue', 'Angelina Violet and

White', 'AngelMist Dark Rose', and 'AngelMist Pink' required the longest length of time to

reach marketability after starting to flower. This information will allow growers to monitor crop









development and schedule shipment of the finished product for approximately 1 week after the

start of flowering.

Production guidelines for angelonia recommend scheduling 7 to 8 weeks for Angelface

cultivars (PW, 2006), 8 to 9 weeks for Angelina cultivars (Selecta, 2008), 7 to 9 weeks for

AngelMist cultivars (BFP, 2006), and 9 to 10 weeks for Serena cultivars (PanAm, 2006) grown

in 2.2 L (1 gal.) pots. However, this length of time may vary from between 6 and 10 weeks

depending upon time of year and greenhouse environment (Schoellhorn and Alvarez, 2002). In

this experiment, crop production time varied from 6.5 to 10 weeks. AngelMist and Serena

cultivars reached marketability within their anticipated crop windows. Angelina cultivars

finished one week earlier and Angelface cultivars finished two weeks later than anticipated.

These variations from the published guidelines may be due to differences in the climate and

environment where cultivars were trialed before commercial release. Angelonia will finish more

quickly when grown under warmer temperatures and higher light, up to an optimum, and the

reverse when grown cooler and under lower light (Miller and Armitage, 2002).

Plant height and width at marketability

Plant height ranged from 14.0 cm for 'AngelMist Basket Pink' to 64.5 cm for 'Angelface

Blue Bicolor' (Table 2-4), and the average height was 39.0 cm. The three shortest were

AngelMist Basket cultivars, a sub-group of cultivars within the AngelMist series with trailing

growth habits rather than the upright or mounding habits observed in most other cultivars (Fig. 2-
4-
1).

Plant width ranged from 28.0 cm for 'AngelMist Dark Lavender' to 54.0 cm for

'Angelface Blue Bicolor' and the average width was 41.5 cm. 'Angelface Blue Bicolor' was

both the tallest and widest cultivar. It would have had a much narrower width except for the fact

that it began to lodge in the latter stages of production. It was also the latest cultivar to flower









and thus had the longest number of days to grow before data collection. The other very wide

cultivars had mounding to semi-trailing growth habits, for example in 'AngelMist Basket Pink'

and 'Angelina Pink and White'. The narrowest cultivars can be grouped into two categories: 1)

cultivars that flower extremely early and thus did not have as much growing time before data

collection, such as 'AngelMist Dark Lavender', and 2) cultivars with very tall, upright growth

habits, such as 'Angelface Pink'.

The growth habit of a plant is an important consideration during crop production. If a

plant is extremely tall or wide, it will be difficult to grow it successfully. Plants with tall growth

habits will not fit well on shipping racks and plants with wide, sprawling growth habits will be

difficult to grow on tight spacing in a greenhouse. Cultivars with one or both of these

characteristics will likely benefit from the application of a plant growth regulator during

production. However, these are traits which a breeder may be able to select against when

developing new cultivars.

Inflorescence number at marketability

At marketability, the cultivars with the least and greatest number of inflorescences were

'Angelina Violet and White' and 'AngelMist Lavender' with 2.3 and 4.2, respectively (Table 2-

4). The average number of inflorescences across all cultivars was 3.3. This measurement gives

an idea of the appearance of the plant at marketability. Cultivars that have fewer inflorescences

with open flowers at marketability have a couple of well-developed inflorescences. Cultivars

with more inflorescences at marketability have fewer open flowers per inflorescence. Growers

will require longer production times to produce cultivars like 'Angelface White' and will require

a longer period of time for it to develop a sufficient number of inflorescences and a good display

of color. However, the one advantage seen in 'Angelface White' relative to 'AngelMist Dark









Lavender' is that it has larger flowers on its inflorescences and will require fewer flowers to

produce a similar display of color.

Inflorescence number at 9.5 weeks

The number of inflorescences per plant was determined at 9.5 weeks following pinch.

This is different from the determination of the number of inflorescences at market because this

will give an idea of its display of color after a specified point in time. The range for number of

inflorescences per plant at 9.5 weeks after pinch was from 2.1 for 'Angelface Dresden Blue

Bicolor' to 29.2 for 'AngelMist Dark Lavender' (Table 2-5), and the average was 12.6

inflorescences. Both time to flower and genotype are important components that help determine

the number of inflorescences with open flowers at a specific point in time. Cultivars that

naturally take longer to flower, such as 'Angelface Pink' (refer to Table 2-3) will have fewer

inflorescences in color than cultivars that naturally flower earlier, such as 'AngelMist Dark

Lavender' given the same set of environmental conditions (Fig. 2-2). Also, the taller cultivars

that have naturally upright growth habits, such as 'Angelface White' and 'Angelface Purple

Stripe', tend to have fewer inflorescences per plant than those with compact or spreading growth

habits, such as 'AngelMist Dark Lavender' and 'AngelMist Lavender Stripe.' Cultivars that

have naturally tall, narrow growth habits and take longer to flower, such as 'Angelface White',

relative to compact, early-flowering cultivars, such as 'AngelMist Dark Lavender', have the

undesirable characteristics for both traits.

Flower size

Flower size was measured at 9.5 weeks following pinch. Flower height and flower width

are positively related to one another, and taller flowers tend to have larger widths than shorter

flowers. Flowers, in general, are round or oval in shape (Fig. 2-3). In certain flowers, the side

petals reflex backward more than others. This backward bending of the flower causes a decrease









in overall flower width and decreases the amount of visual impact created by the flower. Larger

flowers tend to have a rounded shape and smaller flowers tend to have an oval shape. Very little

petal reflex is observed in the Angelface series and the AngelMist Basket cultivars. Medium

sized flowers, such as those on 'AngelMist Purple Stripe', tend to also be round in shape, but

will have a slight more degree of petal reflex than seen in the largest-flowering cultivars.

Cultivars with small flowers, such as the Serena series or 'AngelMist Dark Lavender', tend to

exhibit the greatest degree of petal reflexing.

Flower height ranged from 20.1 mm in 'AngelMist White Improved' to 30.7 mm in

'Angelface Dresden Blue'. Flower width ranged from 12.7 mm in 'AngelMist Pink' to 28.4 mm

in 'Angelface Blue'. Average flower height and width were 23.8 and 20.0 mm, respectively.

Cultivars in the Angelface series had the tallest and widest flowers. Cultivars in the other three

series did not group together very well, but all of the Serena cultivars had flowers shorter and

narrower than the overall averages (Fig. 2-4).

In general, cultivars with tall growth habits tend to have larger flowers than the cultivars

with compact growth habits. Exceptions include the AngelMist Basket cultivars, with trailing

growth habits, and 'AngelMist Purple Improved', which has a medium growth habit but

relatively large flowers.

Powdery mildew index

Powdery mildew susceptibility was not one of the initial traits of interest in this

experiment. However, during the cultivar screen, it was a natural occurrence in the greenhouse,

and was added as a trait of interest since cultivars exhibited varying levels of sensitivity. Plants

were not inoculated for this study, and no fungicides were applied for the duration of the

experiment so that the degree of susceptibility could be quantified. Plants were evaluated on a 1-

5 rating at the end of the experiment, and susceptibility ranged from highly resistant to highly









susceptible. The most susceptible cultivar, with an average rating of 5.0, was 'AngelMist White

Improved' (Table 2-5). The least susceptible cultivars, with ratings of 1.0, were 'Angelface

Blue', 'Angelface Blue Bicolor', 'Angelina Violet and White', 'AngelMist Basket Pink',

'AngelMist Pink', 'AngelMist Dark Rose', and 'Serena Lavender Pink'.

Over half of the cultivars in this study had ratings less than 2.0, which would be

considered saleable. Anything with greater than a 2.0 rating would have benefited from a

fungicide spray in order to retard the spread of the pathogen from the lower leaves up through

the plant canopy. Cultivars with a high degree of susceptibility should be closely monitored

during production to prevent an outbreak from affecting plant quality. The range of

susceptibility observed indicates that powdery mildew tolerance might be a trait that breeders

could select for in cultivar development.

Experiment 2-2

Days to first flower

The season x cultivar interaction was significant for number of days to first open flower

and number of days to marketability (Table 2-6). The mean separations for each parameter were

very similar, so the discussion will focus on number of days to first flower.

Generally, the number of days to first open flower decreased as the seasons progressed

from late winter (season 1) to early summer (season 4) (Table 2-7). The difference in days to

flower between season 1 and season 4 was the least (17 d) in 'AngelMist Dark Lavender' and the

greatest (almost 29 d) in 'Angelface Blue'. This is a difference of 2 to 4 weeks in production

from late winter to early summer.

'AngelMist Dark Lavender' was the quickest to flower in all four seasons. 'Angelface

White' and 'AngelMist Purple Improved' were the slowest to flower in seasons 1 and 2, and

'Angelface White' was the slowest in seasons 3 and 4. The range between the first and last









cultivar to flower was smallest in season 4, at 16.2 days. A 2-week difference in production time

can greatly impact the number of turns in a greenhouse during the spring production season if a

late-flowering cultivar is grown instead of an early-flowering cultivar. This additional

production time may be justified, however, if it is a unique cultivar and can be sold at a higher

price.

Temperature and light, both photoperiod and irradiance, has been shown to play

important roles in flower initiation and development. Angelonia is a day-neutral species (Miller

and Armitage, 2002; Starman, 2001) so irradiance and total light are more important variables

than photoperiod for this crop. Irradiance plays a role in floral initiation, and increased

irradiance, up to an optimum, will hasten floral initiation (Kaczperski et al., 1991). As daylength

increases, the DLI, or total irradiance captured in a 24-h period, will increase due to a longer day

length from which to capture light energy (Korczynski et al., 2002). Temperature is involved in

the rate of development of the floral buds. Since floral development is a metabolically-

controlled process, an increase in temperature will hasten development up to an optimum, and

then retard development at supraoptimal temperatures (Kaczperski et al., 1991). Increased

irradiance hastened flowering by 5 d in Achillea and 7 days in Gaura when grown at a constant

22 C (Fausey et al., 2005). Increased average daily temperature hastened flowering in

Campanula carpatica 'Blue Clips' (Nui et al., 2001), and pansy 'Universal Violet' exhibited a

linear increase in the rate of progress to flowering as temperature increased up to an optimum of

21.7 C, resulting in earlier flowering at higher temperatures (Adams et al., 1997). Increases in

both temperature and irradiance hastened flowering in two Gypsophilapaniculata cultivars

(Hinkleton et al., 1993) and in Pelargonium xhortorum 'Radio' (Welander, 1983).









These results for angelonia are consistent with results from Miller and Armitage (2002), in

which the number of days to visible bud and days to flower decreased with an increase in either

irradiance or temperature. In their work each variable was examined in separate experiments so

it is unknown if there is an interaction between temperature and irradiance in angelonia. For the

work reported here, the average daily temperature increased from 21.4 C in season 1 to 28.0 C

in season 4. The daily light integral (DLI) increased from 20.2 mol-m-2d-1 in season 1 to 21.5

mol-m-2d-1 in season 2, then declined to 20.6 and 17.9 mol-m-2d-1 in seasons 3 and 4,

respectively. Although irradiance declined in seasons 3 and 4, the number of days to flower still

decreased. This may indicate that temperature has more influence on flowering than light above

a minimum light level. The light levels observed in this experiment (17.9 to 21.5 mol-m-2-d-1)

were relatively high compared to those observed in greenhouses in northern states in early spring

(typically less than 10 mol-m-2-d-1) (Korczynski et al., 2002) and may not have been the critical

factor influencing flower time.

Plant height

The cultivar x season interaction was significant, and the general trend observed was an

increase in plant height at marketability from season 1 to season 4 in all cultivars (Table 2-8).

'AngelMist Purple Improved' had the least difference in height between seasons (7 cm) and was

the only cultivar in which plant height was not significantly different across season. 'AngelMist

Purple Stripe' had the greatest increase in plant height between seasons (26 cm).

Within the different seasons, 'AngelMist Dark Lavender' was the shortest cultivar in

every season. 'AngelMist Purple Stripe', 'Angelface White', and 'Angelface Blue' were the

tallest cultivars in season 1, and 'AngelMist Purple Stripe' and 'Angelface White' were the

tallest in seasons 2 through 4. The range between the tallest and shortest cultivars was 33 to 35

cm in seasons 1 through 3, but increased to 45 cm in season 4.









In the study conducted by Miller and Armitage (2002), looking at the effects of

increasing temperature or irradiance separately, it was reported that an increase in temperature

had a greater influence on plant height than an increase in irradiance. In their experiments, plant

height increased by almost 17 cm when grown at a constant 30 OC compared to 22 C, and plants

grown at 30 C had longer intemodes and more brittle stems than those grown at 22 C. Plant

height in the highest irradiance level was only 3 cm shorter than those grown at the lowest level.

An increase in both temperature and irradiance should have opposing influences on plant height,

but temperature appears to have a greater influence on final plant height than irradiance,

especially under higher light levels. Increased plant growth in the later seasons will require

growers to use additional height control measures.

Flower size

The analysis for this parameter was based on only six cultivars due to missing data for

'Angelface White' in season 4. At the time of data collection, not enough plants had mature,

open flowers for accurate measurement of the flower height. Since the results for flower height

and flower width were very similar, only the results for flower height will be presented.

The interaction was not significant, but the main effect of season and the main effect of

cultivar were significant. The lack of an interaction indicates that flower size in all cultivars

responded similarly to environmental changes with season while other growth parameters

showed differences in cultivar responses to season. The differences in flower size for the

cultivars are consistent with the results observed in Expt. 2-1, and is not surprising considering

that the flower size was one parameter used to select the cultivars for this study.

Flower size has been shown to decrease as temperature increases in chrysanthemum

(Willits and Bailey, 2000); calendula, impatiens, mimulus, and torenia (Warner and Erwin,

2005a); and pansy (Warner and Erwin, 2006). However, in this study on angelonia, the range









between seasons was only 1.5 mm, a very small amount. Flower height was greatest in season 1,

but no significant differences were observed between seasons 2 through 4 (Table 2-9).

'Angelface Blue' had the tallest flowers (average of 29.4 mm for the 4 seasons), followed by

'AngelMist Purple Improved'. 'AngelMist Dark Lavender' had the shortest flowers (21.1 mm)

and was significantly different from all cultivars except 'AngelMist Lavender Stripe'.

Powdery mildew susceptibility

The season by cultivar interaction was significant for powdery mildew susceptibility

(Table 2-6). The level of susceptibility observed in Expt. 2-2 is similar to the level of

susceptibility observed for the same cultivars in Expt. 2-1. The level of susceptibility for five of

the cultivars was similar across season, and was low relative to the other cultivars (Table 2-10).

'AngelMist Lavender Stripe' had a greater incidence of powdery mildew in seasons 2 and 4

relative to plants of the same cultivar grown in seasons 1 and 3. 'AngelMist Dark Lavender' had

a greater incidence of powdery mildew in seasons 1 through 3 relative to plants of the same

cultivar grown in season 4. Under these experimental conditions, 'AngelMist Dark Lavender'

was the most susceptible cultivar.

In Florida, powdery mildew tends to be a more common occurrence in the spring months

and less of an issue in the summer months. This is due to the fact that warm and dry

environments are most favorable for spore germination and hyphal growth (Agrios, 2005).

Conclusions

In Expt. 2-1, Angelonia angustifolia cultivars exhibited a wide range of variability in all of

the growth and flowering traits quantified. The number of days to first flower spanned a period

of more than 3 weeks, a long time for a crop with a total production time of 6.5 to 10 weeks.

Cultivars varied in plant height and width at marketability by more than 50 and 26 cm,

respectively. Plants with shorter, wider habits tended to have more inflorescences open at









marketability. At 9.5 weeks after pinch, once all the cultivars had started flowering, those with

wider, spreading habits had more inflorescences than those with narrower, upright growth habits.

Cultivars with narrow, upright growth habits tended to have fewer inflorescences per plant but

larger flowers. Powdery mildew susceptibility was highly variable amongst cultivars and ranged

from highly resistant to highly susceptible.

In Expt. 2-2, number of days to first open flower, plant height, and powdery mildew

susceptibility were all subject to season x cultivar interactions. Flower size was influenced by

the main effects of cultivar and temperature. In the warmest season (season 4, early summer),

plants flowered quickly, were tall, had the least susceptibility to powdery mildew, and had

slightly smaller flowers than plants grown in late winter. Cultivars varied widely in their

response to changing temperature and irradiance levels. 'AngelMist Dark Lavender' did not

flower any quicker between seasons 2 and 4, but other cultivars, like 'Angelface Blue' exhibited

a continual decrease in the number of days to flower with each successive season. These results

indicate that there is variability present between cultivars that may be useful in new cultivar

selection.

The results from Expt. 2-2 indicate that seasonality is an important factor to consider in

commercial production. In southern states, early to late spring would be ideal for production. In

late winter, this crop would need extra time to reach maturity and in early summer, plant quality

would likely decrease. In northern states, the use of supplemental heating and/or lighting will

help hasten crop maturity for the spring season.


























Figure 2-1. Range of plant height present in commercial Angelonia angustifolia cultivars. A)
'AngelMist Purple Stripe' B) 'Angelina Pink' C) 'AngelMist Basket Pink.' Height
data (Table 2-4) collected at marketability was 55.5, 38.5 and 14.0 cm, respectively.
Pictures were taken at 9.5 weeks after pinch, and plant heights were 66.0, 39.0, and
20.5 cm, respectively.

...... .. .........














Figure 2-2. Examples of the range on inflorescence number present in Angelonia angustifolia
cultivars 9.5 weeks after pinching. Plants were grown from January to April 2007.
The cultivars and average number of inflorescences are as follows: A) 'AngelMist
Dark Lavender' with 29.2 inflorescences B) 'Angelina Pink and White' with 15.7 C)
'Angelface Pink' with 2.3.



















Figure 2-3. Range of flower size present in commercial Angelonia angustifolia cultivars. From
left to right, the cultivars are 'Angelface Blue', 'AngelMist Purple Stripe', and
'Serena White'. In many of the small-flowered cultivars, the side petals reflex
backwards to a greater degree than in larger flowers.


Figure 2-4. Comparison between inflorescences in Angelonia angustifolia 'Angelface White'
(left) and 'Serena White' (right). Note differences in inflorescence length, flower
size, and the spacing of the flowers along the inflorescence.










Table 2-1. Plant growth and flowering characteristics for seven Angelonia angustifolia cultivars used in Expt. 2-2.


Growth habit
Plant height
Plant width
Internode length
Branching
Stem thickness
Leaf thickness
Leaf pubescence
Leaf stickiness


Flowering
Flower size
Time to flower
Internode length
Inflorescence number per plant
Flower number per inflorescence
Petal rolling of side petals
Flower color


Angelina
Violet and
White

Medium
Wide
Medium
Medium
Medium
Thin
Glabrous
Medium


Medium
Medium
Medium
Medium
Medium
High
Bicolored -
purple upper
petals and
white lower
petals


AngelMist
Dark
Lavender

Short
Medium
Short
High
Thin
Thin
Glabrous
Low


Small
Early
Short
High
High
High
Lavender


AngelMist
Lavender
Stripe

Medium
Wide
Medium
High
Medium
Thin
Glabrous
Medium


Medium
Medium
Medium
Medium
Medium
Medium
Bicolored -
white petals
with light
lavender
stripe down
center


AngelMist
Purple
Improved

Medium
Medium
Medium
Medium
Medium
Medium
Glabrous
Medium


Large
Late
Medium
Low
Medium
Medium
Purple


AngelMist
Purple
Stripe

Tall
Narrow
Long
Medium
Thick
Thick
Pubescent
Very high


Large
Medium
Long
Low
High
Low
Bicolored -
white petals
with purple
stripe down
center


Angelface Angelface
Blue White


Tall
Narrow
Long
Medium
Medium
Medium
Glabrous
Medium


Large
Medium
Long
Medium
High
Low
Blue with
purple
specks


Tall
Narrow
Long
Low
Thick
Thick
Pubescent
Very high


Large
Late
Long
Low
High
Medium
White










Table 2-2. Temperature and light data for Expt. 2-2.
Season 1 Season 2
Late winter Early spring
(30 Jan 20 Apr) (21 Mar 18 May)
Daily max T (C) 28.0 31.7
Daily min T (C) 17.0 17.7
24-hr average (C) 21.4 23.5


Season 3
Late spring
(25 Apr 20 Jun)
34.4
19.6
25.7


Season 4
Early summer
(6 Jun 23 Jul)
36.2
22.7
28.0


DLI (molm-2-d-1)z 20.2
z DLI = daily light integral


21.5


20.6


17.9









Table 2-3. Angelonia angustifolia flowering data for 31 cultivars (Expt. 2-1). Marketability was
defined as 20 open flowers per plant. Analysis of variance for both parameters were
significant at P<0.001.


Cultivar


Angelface Blue
Angelface Blue Bicolor
Angelface Dresden Blue
Angelface Pink
Angelface Wedgewood Blue
Angelface White

Angelina Blue
Angelina Dark Blue
Angelina Pink
Angelina Pink and White
Angelina Violet and White
Angelina White

AngelMist Basket Pink
AngelMist Basket Purple
AngelMist Basket White
AngelMist Dark Lavender
AngelMist Dark Pink
AngelMist Dark Rose
AngelMist Deep Plum Imp.
AngelMist Lavender
AngelMist Lavender Stripe
AngelMist Pink
AngelMist Plum
AngelMist Purple Imp.
AngelMist Purple Stripe
AngelMist White Cloud
AngelMist White Imp.

Serena Lavender
Serena Lavender Pink
Serena Purple
Serena White

Analysis of variance
Waller-Duncan MSD,=o.05


First open
flower
(days from
pinch)
54.0
63.8
60.8
59.8
56.1
59.4


49.3
42.5
47.4
52.4
49.1
47.9

54.1
52.4
47.4
41.4
52.6
48.3
53.9
47.4
44.6
49.6
57.4
59.6
54.3
44.1
50.7

57.2
56.1
57.2
56.4


2.5


Marketability
(days from
pinch)


58.8
69.8
66.0
66.4
59.9
64.9

56.5
49.4
53.5
57.3
57.0
53.9

58.6
57.7
53.4
45.4
57.0
55.8
57.4
53.0
53.1
57.1
62.8
64.6
58.8
50.4
57.0

63.9
61.4
63.8
62.8


2.4









Table 2-4. Plant data for 31 cultivars ofAngelonia angustifolia collected at marketability (Expt.
2-1). Marketability was defined as the first date at which a plant had 20 or more open
flowers. Analysis of variance for all parameters were significant at P<0.001.
Cultivar Plant height Plant width Inflorescence
(cm) (cm) number
Angelface Blue 52.0 41.5 3.2
Angelface Blue Bicolor 64.5 54.0 3.2
Angelface Dresden Blue 56.0 36.5 3.0
Angelface Pink 48.5 36.5 2.7
Angelface Wedgewood Blue 49.5 36.5 3.0
Angelface White 59.5 39.5 2.6

Angelina Blue 40.0 44.0 3.3
Angelina Dark Blue 31.0 42.0 3.1
Angelina Pink 38.5 39.0 3.5
Angelina Pink and White 42.5 44.5 3.6
Angelina Violet and White 44.0 43.0 2.3
Angelina White 32.0 35.5 3.6

AngelMist Basket Pink 14.0 53.5 3.2
AngelMist Basket Purple 20.0 53.0 3.2
AngelMist Basket White 23.5 47.0 2.7
AngelMist Dark Lavender 26.0 28.0 3.6
AngelMist Dark Pink 39.5 46.5 3.6
AngelMist Dark Rose 35.0 36.5 4.0
AngelMist Deep Plum Imp. 45.5 36.5 2.4
AngelMist Lavender 31.0 36.0 4.2
AngelMist Lavender Stripe 36.0 42.5 3.6
AngelMist Pink 33.5 40.0 3.8
AngelMist Plum 45.0 37.0 3.4
AngelMist Purple Imp. 44.0 38.5 3.2
AngelMist Purple Stripe 55.5 43.5 3.2
AngelMist White Cloud 32.0 36.0 2.9
AngelMist White Imp. 39.0 44.0 3.4

Serena Lavender 34.5 41.0 3.8
Serena Lavender Pink 33.0 41.0 3.9
Serena Purple 32.5 39.5 4.0
Serena White 36.0 47.0 3.9

Analysis of variance *** *** ***
Waller-Duncan MSD, o.05 3.5 4.5 0.8









Table 2-5. Angelonia angustifolia plant data for 31 cultivars collected 9.5 weeks after pinch
(Expt. 2-1). Powdery mildew was rated from 1 to 5, with 1 = least incidence of
powdery mildew and 5 = most severe incidence of powdery mildew. Analysis of
variance for all parameters was significant at P<0.001.
Cultivar Number of Flower height Flower width Powder
inflorescences (mm) (mm) mildew in
Angelface Blue 4.2 30.3 28.4 1.0
Angelface Blue Bicolor 2.1 24.6 23.2 1.0
Angelface Dresden Blue 2.9 30.7 27.3 1.1
Angelface Pink 2.3 26.4 24.5 1.1
Angelface Wedgewood Blue 4.1 28.7 25.9 3.6
Angelface White 2.3 29.4 26.3 1.1


Angelina Blue
Angelina Dark Blue
Angelina Pink
Angelina Pink and White
Angelina Violet and White
Angelina White

AngelMist Basket Pink
AngelMist Basket Purple
AngelMist Basket White
AngelMist Dark Lavender
AngelMist Dark Pink
AngelMist Dark Rose
AngelMist Deep Plum Imp.
AngelMist Lavender
AngelMist Lavender Stripe
AngelMist Pink
AngelMist Plum
AngelMist Purple Imp.
AngelMist Purple Stripe
AngelMist White Cloud
AngelMist White Imp.

Serena Lavender
Serena Lavender Pink
Serena Purple
Serena White

Analysis of variancey
Waller-Duncan MSD-=o.05


17.3
16.1
19.6
15.7
20.3
18.6

8.7
10.9
12.2
29.2
11.6
19.2
5.6
27.6
24.7
21.1
4.1
2.9
4.8
26.9
24.3

5.3
8.2
7.2
9.7

***
3.4


22.0
24.5
21.6
21.7
22.6
24.1

23.6
26.3
24.3
20.7
25.3
22.6
23.7
21.0
22.5
21.9
22.5
25.0
24.7
21.1
20.1

21.7
22.6
22.0
20.6


1.6


y
dexz


18.8
21.0
18.7
19.6
19.6
16.6

21.0
24.3
21.3
18.4
19.3
14.5
22.1
15.3
19.3
12.7
13.2
22.6
22.1
14.6
14.4

19.6
19.1
18.0
17.1


1.4









Table 2-6. Split plot analysis of variance for seven Angelonia angustifolia cultivars grown at four different seasons from January to
July 2007 (Expt. 2-2). Plant height was collected as each plant reached marketability. Powdery mildew index and flower
height data were collected on the same day once all cultivars reached marketability.
Days to first Days to Plant height Powdery Flower height
flowery marketability (cm) mildew index (mm)
Season **
Cultivar *** *** *** *** ***
Seas. x Cv. *** *** *** *** NS
y NS *, **, and *** non-significant or significant at P<0.05, 0.01, and 0.001, respectively.

Table 2-7. Number of days to first flower in seven Angelonia angustifolia cultivars across four seasons (Expt. 2-2). The dates for
each season were the following: late winter 30 Jan to 20 Apr; early spring 21 Mar to 18 May; late spring 25 Apr to 20
Jun; and early summer 6 Jun to 23 July 2007. Uppercase letters indicate mean separation within each column (between
seasons within each cultivar) and lowercase letters indicate mean separation within each row (between cultivars within
each season).


Season
1 Late winter
2 Early spring
3 Late spring
4 Early summer


Angelina
Violet and
White
49.1 Ac
41.3 Bbc
35.7 Cc
28.8 Dbc


AngelMist
Dark
Lavender
41.4 Ad
26.3 B e
27.6 B e
24.2 B d


AngelMist
Purple Imp.

59.6 A a
46.8 B a
39.5 Cb
32.3 Db


Cultivar
AngelMist
Lavender
Stripe
44.6 Ad
38.3 B cd
31.8 Cd
26.1 D cd


AngelMist
Purple Stripe

54.3 Ab
41.9 Bb
39.5 Bb
32.1 Cb


Angelface
Blue


Angelface
White

59.4 A a
49.5 B a
46.9 Ba
40.4 C a


54.0
36.4
29.5
25.2


Ab
Bd
C de
Dd


I









Table 2-8. Plant height (cm) at marketability in seven Angelonia angustifolia cultivars across four seasons (Expt. 2-2). Marketability
was defined as the first date at which a plant had 20 or more open flowers. The dates for each season were the following:
late winter 30 Jan to 20 Apr; early spring 21 Mar to 18 May; late spring 25 Apr to 20 Jun; and early summer 6 Jun
to 23 July 2007. Uppercase letters indicate mean separation within each column (between seasons within each cultivar)
and lowercase letters indicate mean separation within each row (between cultivars within each season).
Cultivar
Angelina AngelMist AngelMist AngelMist AngelMist Angelface Angelface
Violet and Dark Purple Imp. Lavender Purple Stripe White Blue
Season White Lavender Stripe
1 -Late winter 44.0 B b 26.0 B d 44.0 Ab 36.0 B c 55.5 C a 59.0 C a 52.0 AB a
2 Early spring 50.0 AB b 32.5 AB c 48.5 Ab 46.5 Ab 63.0 BC a 67.0 B a 51.5 B b
3 -Late spring 52.0 A bc 38.0 A d 47.0 Ac 45.5 Ac 69.0 B a 73.5 AB a 58.0 AB b
4 Early summer 53.0 A bc 37.0 A d 51.0 Ac 51.0 Ac 81.5 A a 78.0 A a 59.0 A b

Table 2-9. Flower height (mm) in six Angelonia angustifolia cultivars across four seasons (Expt. 2-2). Data was collected once all
cultivars reached marketability. 'Angelface White' had missing data for the early summer season and was non-estimable,
creating an unbalanced factorial. Data were analyzed for the remaining six cultivars with complete data sets for all
seasons. The dates for each season were the following: late winter 30 Jan to 20 Apr; early spring 21 Mar to 18 May;
late spring 25 Apr to 20 Jun; and early summer 6 Jun to 23 July 2007.
Cultivar
Angelina AngelMist AngelMist AngelMist AngelMist Angelface Pooled
Violet and Dark Purple Imp. Lavender Purple Blue mean across
Season White Lavender Stripe Stripe cultivar
1 Late winter 22.6 23.0 25.0 22.5 24.7 30.3 24.7 a
2 Early spring 22.1 21.0 24.3 21.1 23.4 29.7 23.6 b
3 Late spring 21.3 20.0 24.2 20.8 22.6 28.6 22.9 b
4 Early summer 21.3 20.4 23.6 20.3 22.3 29.1 22.9 b


Pooled mean by
cultivar


24.3 b 21.2 de 23.3 c


21.8 d 21.1 e


29.4 a









Table 2-10. Powdery mildew index in seven Angelonia angustifolia cultivars across four seasons (Expt. 2-2). The index ranged from
1 to 5, with 1 indicating the least incidence of powdery mildew and 5 indicating a severe incidence of powdery mildew.
The dates for each season were the following: late winter 30 Jan to 20 Apr; early spring 21 Mar to 18 May; late spring
25 Apr to 20 Jun; and early summer 6 Jun to 23 July 2007. Uppercase letters indicate mean separation within each
column (between seasons within each cultivar) and lowercase letters indicate mean separation within each row (between
cultivars within each season).


Season
1 Late winter
2 Early spring
3 Late spring
4 Early summer


Angelina
Violet and
White
1.0 Ac
1.1 Ab
1.2 Ac
1.0 Ab


AngelMist
Dark
Lavender
4.7 Aa
5.0 Aa
4.6 Aa
1.9 Ba


AngelMist
Purple Imp.

1.3 Ac
1.4 Ab
1.2 Ac
1.0 Ab


Cultivar
AngelMist
Lavender
Stripe
2.6 Ab
1.7 Bb
2.2 Ab
1.0 Bb


AngelMist
Purple Stripe

1.1 Ac
1.3 Ab
1.2 Ac
1.0 Ab


Angelface
White

1.1 Ac
1.0 Ab
1.0 Ac
1.0 Ab


Angelface
Blue

1.0 Ac
1.0 Ab
1.5 Ac
1.0 Ab









CHAPTER 3
PLANT DATE X CULTIVAR INTERACTIONS INFLUENCE SUMMER LANDSCAPE
PERFORMANCE OF Angelonia angustifolia



Introduction

High temperature stress in plants can lead to a reduction in flower number, an increase in

floral bud abortion, or complete cessation of flowering. The mechanism underlying this

temperature response may be related to either acute or chronic high temperature stress depending

upon species. In greenhouse production, decreased flower bud number has been observed in

pansy and petunia (Kaczperski et al., 1991; Warner and Erwin, 2006) In field crops, high

temperature stress has been shown to reduce flower number in Capsicum annuum (Erickson and

Markhart, 2001) and reduce yield in tomato and canola (Morrison and Stewart; Sato et al., 2000).

Warner and Erwin (2005b) published a model for floral abortion in Arabidopsis thaliana

genotypes and indicated that plant response was related to a threshold number of hours above a

critical temperature.

For landscape beds and other large-scale plantings, it is critical to select plants that will

flower throughout the season. Angelonia angustifolia is a plant with reported heat tolerance but

has been observed to go out of flower during the summer months in Florida (personal

observation). Different cultivars will stop flowering at different times during the summer and

remain without flowers for varying lengths of time. The objective of this research was to

determine the effect of plant date and cultivar on landscape performance and flowering of eight

angelonia cultivars.

Materials and Methods

See Experiment 2-2 for a description of materials and methods related to the production of

rooted cuttings. This experiment consisted of the seven cultivars shown in Table 2-1 and 'Serena









White' planted at three different times into a landscape bed. Rooted cuttings of all cultivars were

planted into 80 mm Ellepots (Knox Nursery, Winter Park, Florida) on 14 Mar., 18 Apr., and 16

May 2007, respectively, and grown in a natural ventilated greenhouse for 3 weeks. They were

pinched two weeks after planting, and twelve plants of each cultivar were transplanted into an in-

ground landscape bed (Gainesville, FL) on three plant dates spaced at 4-week intervals, on 4

-2
Apr., 2 May, and 30 May 2007, respectively. The bed was amended with 122 gm-2 Osmocote

18-6-12 (18.0N-2.6P-10.0K) slow-release fertilizer with 6 to 9 month release rate (The Scotts

Co., Marysville, OH), and plants were irrigated as needed with drip irrigation. This experiment

was designed as a split plot, with plant date as the main plot and cultivar as the subplot. Within

each plant date, cultivars were planted in a complete random block design consisting of three

blocks and four plants per block. Plants were planted 46 cm apart within each row and 61 cm

between rows.

Data were collected at 10 weeks after transplant (WAT) on two plants of each cultivar per

block (13 Jun., 11 July, and 8 Aug., respectively) and included plant height, inflorescence

number, and flower height. Plant height was measured from the soil surface to the tallest point

of the plant. Inflorescence number was determined by counting all inflorescences containing at

least one open flower. Flower height was measured with calipers. Beginning 5 WAT, plants

were evaluated every two weeks and a flower rating of 0-10 was determined for each row since

all plants within each cultivar row flowered uniformly. A flower rating of 3 was determined to

be the minimum critical value for acceptable landscape performance. The final evaluation date

for all plants was 10 Oct. Temperature and DLI was recorded every 15 min. by a weather station

(Hortimax USA, Inc., Rancho Santa Margarita, CA) and daily average minimum, maximum, and

24-h temperatures and DLI were calculated. Monthly averages can be found in Table 3-1.









Data were analyzed using SAS 9.1 (SAS Institute, Cary, NC) using the PROC MIXED

procedure. The flower ratings were analyzed two different ways: 1) a comparison based upon

plant age, and 2) a comparison based upon the date when evaluations were collected.

Evaluations based upon plant age were used to determine if plants stopped flowering at a similar

physiological age, and evaluations based upon date collected helped determine the amount of

flowering present in cultivars at the same point in time.

Results and Discussion

10-Week Data

Plant height

The plant date x cultivar interaction was significant for plant height as measured 10 weeks

after transplant (WAT) (Table 3-2). Plants were evaluated on 13 Jun., 11 July, and 8 Aug. 2007,

respectively. This evaluation interval was chosen because it provided adequate time for all

cultivars to begin flowering (see Chapter 2). 'Angelina Violet and White' and 'AngelMist

Purple Improved' were the only two cultivars to have significant height differences between

plant dates (Table 3-3). In these two cultivars, plant height increased from plant date 1 (PD 1) to

PD 2, then decreased from PD 2 to PD 3, but only PD 2 and PD 3 were significantly different

from each other.

The difference in height between plant dates can be attributed to a combination of factors,

including temperature and the number of inflorescences per plant. In PD 1, average temperatures

were cooler than in PD 2 and PD 3, so the plants developed slower. Also, angelonia plants have

been shown to develop longer intemodes under higher temperatures (Miller and Armitage,

2002). In PD 3, these two cultivars, 'Angelina Violet and White' and 'AngelMist Purple

Improved', had very few inflorescences per plant, and the inflorescence can represent a

significant portion of the total plant height due to its raceme architecture (Chapter 1). When a









cultivar has no inflorescences, or very few developed ones, the plant will have a shorter overall

plant height than plants with well-developed inflorescences.

In PD1, 'AngelMist Purple Stripe' and 'Angelface White' were taller than all other

cultivars except for 'Angelface Blue'. In PD2 and PD3, 'AngelMist Purple Stripe' and

'Angelface White' were the tallest cultivars. Cultivars that are tall in production will tend to be

tall landscape plants and compact plants in production will be compact plants in the landscape.

Inflorescence number

The plant date x cultivar interaction was significant for inflorescence number (Table 3-4).

'Angelina Violet and White', 'AngelMist Dark Lavender', and 'AngelMist Lavender Stripe' had

significantly less flowers in PD 2 and 3 relative to PD 1. 'Serena White' and 'AngelMist Purple

Improved' had significantly less flowers in PD 3 compared to PD 1 and 2. 'AngelMist Purple

Stripe', 'Angelface White', and 'Angelface Blue' showed no significant difference in

inflorescence number across plant dates. 'Serena White' had the highest number of

inflorescences in PD 1 and 2, and 'Angelface Blue' and 'AngelMist Purple Improved' had the

least number of inflorescences, respectively, in PD 1 and 2. In PD 3, there were no significant

differences between cultivars.

These results indicate that all cultivars except 'AngelMist Purple Stripe', 'Angelface

White', and 'Angelface Blue' had a lower number of inflorescences at the later plant dates and

were more sensitive to the increase in temperature. In addition to having fewer inflorescences,

the plants also had fewer open flowers per inflorescence in the later plantings (data not

presented). It has been reported that plants subjected to high temperature stress will have fewer

flowers than those not subjected to it (Morrison and Stewart, 2002; Sato, 2000). Although

'Angelface Blue' did not have a significant difference between plant dates, it never had a large









display of color relative to the other two cultivars that were unaffected by plant date. 'Angelface

Blue' had 11, 5, and 1 inflorescences, respectively, across plant dates.

The greatest amount of variability between cultivars occurred in PD 1, and the least

amount of variability occurred in PD 3. The range between cultivars with the greatest and least

number of inflorescences was 160, 127, and 29 in PD 1 through PD 3 In PD 1, growing

temperatures were ideal for cultivars to flower at their phenotypic potential, and peak flowering

occurred approximately 10 WAT (see discussion below). In successive plantings, the average

daily temperatures increased while the DLI remained relatively consistent. However, in the later

plantings, the increase in temperature resulted in plants in which flowering was inhibited due to

higher than optimal temperatures for continual inflorescence development. A study by Miller et

al. (2001) indicated that the optimal temperature for net photosynthesis in two angelonia

cultivars was approximately 20 C. Above that temperature, gross photosynthesis continued to

increase but was negated by an increase in dark respiration. Under our conditions during the

summer, it can be expected that flowering will be affected by the high average temperatures

observed relative to the optimal temperature for photosynthesis in this species.

Flower height

The plant date x cultivar interaction was significant for flower height (Table 3-5). The

only cultivar with a significant change in flower size across plantings was 'AngelMist Purple

Improved'. The mean flower height for this cultivar in each PD were 21.5, 16.6, and 17.9 mm,

respectively, with PD2 significantly smaller than PD 1.

In PD 1, 'Angelface Blue' had taller flowers than all cultivars except 'Angelface White'.

In PD 2, 'Angelface Blue', 'Angelface White', and 'AngelMist Purple Stripe' had the tallest

flowers, and in PD 3, 'Angelface White' had the tallest flowers. The range between the largest

and smallest flowers was 8 to 9.2 mm between plant dates.









In many species a decrease in flower size has been observed with an increase in

temperature, including chrysanthemum, calendula, impatiens, mimulus, torenia, and pansy

(Warner and Erwin, 2005a, 2006; Willits and Bailey, 2000). In angelonia, it appears that the

primary response to high temperature is a decrease in the number of inflorescences and not a

decrease in flower size, whereas in pansy, the tradeoff under high temperatures is a similar

number of flowers, but a decrease in flower size (Warner and Erwin, 2006).

Flower Ratings by Plant Age

Plants were evaluated every two weeks, starting at 5 WAT, for degree of flower coverage

using a 10 point scale, with 0 indicating no inflorescences and 10 indicating 100 percent flower

coverage. A rating of 3 was considered to be the minimum value for acceptable landscape

flowering. Ratings were compared 1) on the basis of plant age and 2) by evaluation date. A

comparison based upon plant age (similar WAT) allows for a comparison of cultivars at the same

physiological age, but exposed to different environmental conditions. At each WAT

comparison, successive plantings were grown under higher average temperatures.

Plants ratings were analyzed from 5 to 19 WAT and the plant date by cultivar interaction

was significant for all intervals except for 19 WAT (Table 3-6). A graphical representation of

each cultivar for the 3 plant dates is given in Figure 3-1. Cultivars varied in their response to the

environmental conditions. For example, 'AngelMist Dark Lavender', a heat-sensitive cultivar

had ratings of 7.7, 2.7, and 1.3 at 9 WAT for PD 1 to 3. In contrast, 'AngelMist Purple Stripe', a

heat-tolerant cultivar had ratings of 6.0, 5.0, and 4.7, respectively. Other cultivars, such as

'AngelMist Purple Improved' had poor landscape performance regardless of plant date.

In all cultivars, the highest flower ratings achieved by each cultivar occurred within PD 1.

In PD 1, cultivars tended to maintain peak flowering for a longer period of time than those from









PD 2 or 3. An exception to this trend was 'AngelMist Purple Stripe' in which all plant dates

maintained flower ratings >3 for all evaluations after 5 WAT.

In PD 1, most cultivars reached peak flowering around 11 WAT and then ratings

generally declined after that. Plants in PD 3, however, never reached a peak flowering window,

and maintained a relatively low amount of flowering (less than or equal to 4) all summer.

Regardless of the actual ratings, all plant dates reach a maximum flowering around 3 months

after transplanting into the gardens, which coincides with their designation as an annual crop.

However, the temperature during initial establishment will have a major impact on the flowering

capability of a cultivar, with higher temperatures inhibiting inflorescence development.

Flower Ratings by Evaluation Date

Flower ratings were also compared by evaluation date. The first evaluation date for all

three plantings was 4 July, once PD 3 had been in-ground for 5 weeks, and ended on 10 Oct.

This provided a comparison of how the different plant dates responded in the landscape at

specific points in time, regardless of plant age. The plant date by cultivar interaction was

significant on 4 July, 18 July, and 29 Aug. (Table 3-7). For the weeks in which the interaction

was not significant, the main effect of cultivar was significant but the main effect of plant date

was not. Cultivars differed significantly in their response to high temperatures.

The interactions observed in 4 and 18 July were due to some cultivars having variable

rating across plant dates while others had consistent ratings. 'Serena White', 'Angelface White',

and 'AngelMist Purple Stripe' had variable ratings (Figure 3-2) due to the fact that PD 3 was

continuing to increase in inflorescence number and PD 1 and 2 still had good flower coverage.

In the other cultivars, however, the high temperatures had inhibited flowering and ratings were

consistently low for all three plant dates. On 29 Aug., the deviation from the general trend was

due to 'Serena White' in PD 3 finally reaching its peak flowering, which coincided with reduced









flowering in the other two plant dates and no variability in other cultivars with respect to plant

date.

In general, cultivars were either heat tolerant or heat sensitive, regardless of plant date.

Temperature throughout the season impacted the amount of flowering observed in each cultivar.

'AngelMist Purple Stripe' had the most consistent flowering and never had ratings <3.0 after 18

July. 'AngelMist Purple Improved', in comparison, did not have any ratings >2.0 all summer.

The decrease in flowering during the summer months is most likely due to changes in

temperature and not changes in DLI. Monthly DLI levels were 35, 37, and 30 mol-m-2-d- for

July, Aug., and Sept. respectively, and average temperatures were 26.8, 27.7, 25.7 C. A

decrease in summer flowering and yield has been reported in canola at temperatures above 29.5

C (Morrison and Stewart, 2002) and at 33 C in tomato (Sato et al., 2000). Warner and Erwin

(2005b) observed in Arabidopsis thaliana that floral buds began to abort at between 200 and 300

h of 33 C, and complete inflorescence failure occurred above 300 h of 33 C. In petunia,

Kaczperski et al. (1991) observed flower abortion at temperatures above 25 C. In our study,

many of the inflorescences formed flower buds but they failed to fully develop and open. On

existing inflorescences, flowers opened sporadically along its length and were frequently

separated by multiple sets of aborted buds.

This decline in flowering is consistent with a decrease in net photosynthesis levels

observed in angelonia at temperatures above 21 C, with 'AngelMist Purple Stripe' having a

higher optimal temperature than 'AngelMist Deep Plum' (Miller et al., 2001). This elevated net

photosynthesis level may possibly indicate that it can tolerate higher temperatures better than

other cultivars.









Conclusions

Angelonia angustifolia cultivars differed in their flowering response to summer conditions.

Plant height, inflorescence number, and flower size were affected by plant date x cultivar

interactions. Inflorescence number in all cultivars decreased with each successive planting, but

the magnitude of the difference was cultivar dependent. For example, 'Serena White' had a

reduction in inflorescence number of 83 percent between PD 1 and 3 while 'AngelMist Purple

Stripe' had only a 53 percent decline.

Plant date 1 had the highest peak floral display value and a longer sustained length of

flower display relative to the later plantings. The decline in flowering was primarily influenced

by chronic high temperature stress during the summer months and not plant age. For spring

landscape plantings (April and early May) in North Florida, all cultivars will provide an

acceptable amount of flower coverage. For plantings after late May, only 'AngelMist Purple

Stripe' will likely provide acceptable flower coverage.
























.6 -


S4-

2e



5 7 9 11 13 15 17 19
Weeks after planting


4 -



0 -
5
5


7 9 11 13 15 17 19
Weeks after planting


".



E0 r -

5 7 9 11 13 15 17 19
Weeks after planting


5 7 9 11 13 15 17 19
Weeks after planting

Plant date 1 Plant date 2 Plant date 3 r n m rn mum flowerna


S


4

0




5


7 9 11 13 15 17 19
Weeks after planting


r




5 7 9 11 13 15 17 19
Weeks after planting

Plant date 1 Plant date 2 Plant date 3 .... I mn mum flowenng


p.


t


U- -


5 7 9 11 13 15 17 19


5 7 9 11 13 15 17 19


Weeks after planting D Weeks after planting H


Figure 3-1. Flower ratings of Angelonia angustifolia graphed by weeks after transplant (WAT).

The plant dates were 4 Apr., 2 May, and 30 May 2007, and the cultivars are A)

'AngelMist Dark Lavender' B) 'AngelMist Purple Improved' C) 'AngelMist

Lavender Stripe' D) 'AngelMist Purple Stripe' E) 'Serena White' F) 'Angelina Violet

and White' G) 'Angelface Blue' H) 'Angelface White'.


I- Plant date 1 Plant date 2 Plant date 3 .... r


I Plant date 1 -Plant date 2 -- P


-- Plant date 1 -- Plant date 2 --Plant date 3 ....


--Plant date 1 --Plant date 2 -- Plant date 3 ....


- Plant date 1 Plant date 2 Plant date 3 .. ..


- -Plant date 1 -- Plant date 2 -- Plant date 3 ....















I- Pant date 1 --P ant date 2 -- Plant date 3 ....


-0 t dt -P dat 2 -Pat dt
10


2

0 -
4 July 18 July 1 Aug 15 Aug 29 Aug 12 Sept 26 Sept 10 Oct
Date


Plant date 1 Plant date 2 --


2
2- -

0
4 July 18 July 1 Aug 15 Aug 29 Aug 12 Sept 26 Sept 10 Oct
Date


- Plant date 1 Plant date 2 Plant date 3 ....


L


4 July 18 July 1 Aug 15 Aug 29 Aug 12 Sept 26 Sept 10 Oct
Date

lt dt -1 2 --P tdt 3 fowe
101


20.


0 ,- '1 .
4 July 18 July 1 Aug 15 Aug 29 Aug 12 Sept 26 Sept 10 Oct


Plant date 1 Plant date 2 -IF-


Ui




4 July 18 July 1 Aug 15 Aug 29 Aug 12 Sept 26 Sept 10 Oct
Date

I- Plant date Plant date 2 IPlant date 3 .... -n-ur flowering
10


E





,'U


u_

4 July 18 July 1 Aug 15 Aug 29 Aug 12 Sept 26 Sept 10 Oct
Date


Plant date Plant date 2 P


6 *- .


I *
S4-
o


0 A 2
4 July 18 July 1 Aug 15 Aug 29 Aug 12 Sept 26 Sept 10 Oct


0ly y et
4 July 18 July 1 Aug 15 Aug 29 Aug 12 Sept 26 Sept 10 Oct


Date D Date H


Figure 3-2. Flower ratings ofAngelonia angustifolia graphed by date of data collection. The

plant dates were 4 Apr., 2 May, and 30 May 2007. The cultivars are A) 'AngelMist

Dark Lavender' B) 'AngelMist Purple Improved' C) 'AngelMist Lavender Stripe' D)

'AngelMist Purple Stripe' E) 'Serena White' F) 'Angelina Violet and White' G)

'Angelface Blue' H) 'Angelface White'.

















79









Table 3-1. Monthly temperature and DLI data for Gainesville, FL for summer 2007.
Average daily Average daily Average 24-hr Average DLI
maximum minimum temperature (C) (tmol-m-2 d-)


Apr
May
Jun
July
Aug
Sept
Oct (1-10)


temperature (C) temperature (C)
25.7 12.9
29.3 17.4
31.2 21.1
32.3 22.7
33.1 23.5
31.2 21.9
29.1 20.5


19.4
23.1
25.5
26.8
27.8
25.7
24.1


36.5
39.2
37.1
35.2
36.9
30.4
25.8


Table 3-2. Split-plot analysis of variance for Angelonia angustifolia growth and flowering data
collected at 10 weeks after planting for three plant dates. Significant levels for
analysis of variance were P <0.05, 0.01, and 0.001.
Plant height Number of Flower number Flower height
(cm) inflorescences per


inflorescence
***


Plant date
Cultivar
Plant date x cultivar










Table 3-3. Mean separation for plant height (cm) of eight Angelonia angustifolia cultivars collected at 10 weeks after transplant for
three plant dates. Uppercase letters indicate mean separation in each column and lowercase letters indicate mean
separation in each row.
Cultivar
Serena Angelina AngelMist AngelMist AngelMist AngelMist Angelface Angelface
White Violet and Dark Purple Lavender Purple White Blue
Plant date White Lavender Improved Stripe Stripe
4 Apr 2007 (PD 1) 43.5 Ac 56.0 AB bc 47.0 A c 49.2 AB c 54.2 A c 75.8 A a 77.0 A a 68.7 A ab
2May2007 (PD2) 46.7 Ae 63.2 A cd 51.0 A de 55.3 A de 64.3 A cd 81.7 Aa 87.3 Aa 69.5 A bc
30 May 2007 (PD 3) 32.7 Ac 41.8 A bc 44.8 A bc 36.3 B c 56.3 Ab 84.0 Aa 74.5 Aa 55.3 Ab

Table 3-4. Mean separation for number of inflorescences per plant of eight Angelonia angustifolia cultivars collected at 10 weeks
after transplant for three plant dates. Uppercase letters indicate mean separation in each column and lowercase letters
indicate mean separation in each row.
Cultivar
Serena Angelina AngelMist AngelMist AngelMist AngelMist Angelface Angelface
White Violet and Dark Purple Lavender Purple White Blue
Plant date White Lavender Improved Stripe Stripe


4 Apr 2007 (PD 1)
2 May 2007 (PD 2)
30 May 2007 (PD 3)


171 Aa
130 A a
30 Ba


79 A cd
11 Bbc
2 Ba


125 Ab
26 B bc
13 Ba


25 A e
3Ac
1 Ba


111 Abc
10 Bbc
9Ba


60 A de
46 Ab
28 Aa


22 A ef
17 Abc
5 Aa


11 Af
5 Abc
1Aa


Table 3-5. Mean separation for flower height (mm) of eight Angelonia angustifolia cultivars collected at 10 weeks after transplant for
three plant dates. Uppercase letters indicate mean separation in each column and lowercase letters indicate mean
separation in each row.


Plant date
4 April 2007 (PD 1)
2 May 2007 (PD 2)
30 May 2007 (PD 3)


Serena
White

17.2 Ae
16.4 Ab
15.9 Ad


Angelina
Violet and
White
19.6 A cd
15.9 Ab
19.6 A bcd


AngelMist AngelMi
Dark Purple


Lavender
17.5 A de
18.5 Ab
18.4 Ad


Improve
21.5 A
16.6 B
17.9 AB


Cultivar
st AngelMist
Lavender
S Stripe
cd 19.5 A cde
b 16.9 Ab
cd 18.1 A cd


AngelMist
Purple
Stripe
22.3 A bc
21.9 Aa
21.5 Ab


Angelface Angelface
White Blue


25.0 A ab
23.6 A a
25.1 Aa


25.6 Aa
23.3 Aa
22.4 A ab


00









Table 3-6. Split-plot analysis of variance for Angelonia angustifolia flower ratings by plant age (weeks after transplant).
5 WATz 7 WAT 9 WAT 11 WAT 13 WAT 15 WAT 17 WAT 19 WAT
Plant date ** *** *** *** ** **
Cultivar
Plant date x cultivar *** *** *** *** *** *** NS *
z WAT = weeks after transplant

Table 3-7. Split-plot analysis of variance for Angelonia angustifolia flower ratings by evaluation date.
Week 27 Week 29 Week 31 Week 33 Week 35 Week 37 Week 39 Week 41
4 July 18 July 1 Aug. 15 Aug. 29 Aug. 12 Sept. 26 Sept. 10 Oct.
Plant date *** NS NS NS *** NS NS NS
Cultivar *** *** *
Plant date x cultivar *** *** NS NS NS NS NS









CHAPTER 4
CULTIVAR BY FERTILIZER INTERACTIONS AFFECT GROWTH OF Angelonia
angustifolia

Introduction

Angelonia angustifolia is an herbaceous perennial native to the neotropics (USDA, 2006).

Its popularity as a floriculture crop has risen in recent years due to virus indexing of unpatented

plant material and to breeding efforts focused on more compact plants and an increased range of

flower colors (Schoellhorn, 2002). Cultivars vary in growth habit, flower size and color, and

flowering time (see Chapter 2). It is not known whether genotypes also vary in response to level

of fertilization.

Published production guidelines for angelonia vary by company. All recommend

constant liquid feed with a complete fertilizer, but the level of fertilization ranges from N levels

of 150 to 250 mg-L-1 (Ball FloraPlant, 2006; PanAmerican Seed, 2006; Proven Winners, 2006;

Fischer, 2008). Recommended N fertilizer guidelines published in the trade press range from 75

to 200 mg-L-1 (Armitage, 1997; Schoellhorn and Alvarez, 2002; Smith, 2007). It is unknown

whether these differences in recommended levels are due to cultivar, series, or environmental

conditions.

Cultivar differences have been noted with poinsettia (Euphorbia pulcherimma), with

optimal fertilization related to leaf color. Dark-green leaved cultivars require an electrical

conductivity (EC) of 1.5 to 2.0 mmhos-cm-1 (mS-cm-1) using the saturated media extract

procedure (SVME), whereas medium-green leaved cultivars require 2.0 to 2.5 mmhos-cm-1 (Ecke,

III et al., 2004). Temperature also influences fertilizer recommendations. The optimal fertilizer

rate for petunia (Petunia xhybrida) decreases as temperature increases (Kang and van lersel,

2001). It is unknown whether or not cultivars within or across commercial angelonia series will

respond similarly to changes in the level of fertilization. The objectives of this experiment were









to determine the response of six angelonia cultivars to varying levels of fertilization and to

determine the optimal concentration of fertilizer for angelonia production.

Materials and Methods

The six angelonia cultivars, 'Serena White', 'AngelMist Dark Lavender', 'AngelMist

Lavender Stripe', 'AngelMist Purple Stripe', 'Angelface White', and 'Angelface Blue', were

selected for this experiment due to their range of growth habits and time to flower. See Expt. 2-2

for maintenance of stock plants and harvesting of unrooted cuttings. Forty-eight rooted cuttings

of each cultivar were planted on 5 June 2007 into 2.2 L (16 cm diameter) pots filled with Fafard

2 soiless medium (Conrad Fafard, Inc., Apopka, FL), watered with regular water for 1 week.

Plants were not fertilized to prevent a buildup of salts in the medium before plants were

transferred to subirrigation benches on 12 July. Treatments were N at 12.5, 25, 50, 100, 200, or

400

mg-L^using a 20.0N-4.4P-16.6K fertilizer (Peters 20-10-20 Florida Special, The Scotts Co.,

Marysville, OH) (0.08, 0.16, 0.33, 0.65, 1.30, and 2.60 mS-m-1) applied at each irrigation. The

electrical conductivity (EC) of the irrigation water was 0.38 mS-cm-1. The experiment was set up

as a split-plot design, with fertilizer concentration as the main plot and cultivar as the sub-plot.

One bench constituted a fertilizer treatment. Within each bench, cultivars were randomized in a

complete random block design with four blocks and two plants of each cultivar per block. Plants

were subirrigated as needed, every 2 to 3 days for the first 3 weeks and then daily for the

remaining 3 weeks. Over the length of the experiment the average daily minimum, maximum,

and 24-hour temperatures were 22.7, 36.2, and 28.00C, respectively. The average daily light

level was 18 mol-m-2-day1.

The number of days to first open flower were recorded. SPAD readings and leachate pH

and electrical conductivity (EC) were collected after 3 and 6 weeks. Plant height, plant width,









number of inflorescences, and dry weight (g) were collected after 6 weeks. SPAD readings

indicate leaf greenness and were taken using a SPAD 502 Minolta chlorophyll meter (Spectrum

Technologies Inc., Plainfield, IL). Four readings per plant were recorded and averaged for both

the upper leaves and the lower leaves. Readings for the upper leaves were taken on recently

mature leaves and readings for the lower leaves were taken from the bottom 25% of the plant.

Leachate pH and EC were collected using one plant per block for all cultivars on each bench.

Leachate was collected 1 hour after irrigation using the PourThru method (Cavins et al., 2000).

Leachate pH and EC were measured using a hand-held HANNA 98130 combination pH/EC

meter (HANNA instruments, Woonsocket, RI). Plant height was measured from the top of the

pot to the tallest growing tip. Plant width was measured at the widest point in the plant canopy

and then again perpendicular the first measurement. Average plant width was used for analysis.

Plant size was calculated as (plant height + average width)/2. Plants were harvested at the soil

surface, bagged by block, and dried at 70 OC for 48 hrs. Dry weight on a per plant basis was

used for analysis. Data were analyzed using SAS 9.1 (SAS Institute, Cary, NC) PROC MIXED

and then regression analysis where appropriate.

Results and Discussion

Substrate pH and Electrical Conductivity

The interaction and the main effects of cultivar were not significant for pH, but the main

effects of fertilizer concentration were significant at 6 weeks (Table 4-1). As a result, the

regression equation for substrate pH relative to fertilizer concentration was pooled over all

cultivars. As the fertilizer concentration increased, the general trend was a decrease in the pH

value of the leachate. At 6 weeks, pH values ranged from 6.4 with N at 12.5 mg-L-1 to 4.3 at 400

mg-L-1 (Fig. 4-1). The6 week values were very similar to the 3 week results, which ranged from

6.7 at 12.5 mg-L-1 to 4.9 at 400 mg-L-1. These results are consistent with the fact that Peter's 20-









10-20 (20N-4.4P-16.6K) has a potential acidity of 410 lbs CaCO3 equivalent per ton (The Scotts

Company, 2004). Over time the pH of the medium and leachate will decrease, with the rate of

decrease quicker as the fertilizer concentration applied is increased.

The recommended pH range for angelonia is 5.5 to 6.2 (Ball, 2006). This range falls

between the 25 and 50 mg-L-1 treatments in this experiment. In this range, however, plants

exhibited visual chlorotic symptoms. It is unknown why the soil solution pH fell below than the

recommended range for plant growth in those treatments supplied with N at 100 mg-L-1 and

higher. The irrigation water used in this experiment had an alkalinity of 1.2-1.4 meq CaCO3,

which should have provided some buffering capacity against sudden drops in pH.

The fertilizer x cultivar interaction was significant for leachate EC at 6 weeks. As the

fertilizer concentration increased, the general trend was an increase in leachate EC (Fig. 4-2),

with the range between cultivars greater as fertilizer concentration increased. The EC range at

the lowest and highest concentrations, 12.5 and 400 mg-L-1, was 0.6 to 0.7 and 3.5 to 6.2

mS-cm-1, respectively. All cultivars except 'Angelface White' and 'AngelMist Purple Stripe'

exhibited a quadratic response (Table 4-2). These two cultivars, which exhibited a linear

response, are the only two cultivars in this experiment with thick, very pubescent, glandular

leaves. The other cultivars had thinner, glabrous leaves. Whether or not these phenotypic

differences are indicators of additional tolerance or sensitivity to fertilization is unknown since

their growth responses were similar to the other cultivars in the range of fertilizer concentrations

tested (see discussion below).

The recommended EC range for angelonia is 0.6 to 0.9 mS-cm-1 using the 2:1 dilution

method (Proven Winners, 2006), which is approximately equivalent to 2.0 to 3.2 mS-cm-1 using









the PourThru method (Cavins et al., 2000). This range falls between the 100 and 200 mg-L-1

treatments, which is consistent with optimal plant growth and flowering (see below).

Leaf Greenness

SPAD readings, a nondestructive measure of leaf greenness, are strongly correlated

(greater than r2=0.9) with actual leaf chlorophyll content obtained using destructive sampling

(Uddling et al., 2007; Wang et al., 2005; Monje and Bugbee, 1992). A measure of the

chlorophyll content in leaves is a strong indicator of the N status of the plant. Although this

experiment did not focus on the effects of increasing N fertilization, but rather increasing levels

of a complete fertilizer, the SPAD meter can be a useful tool for measuring the leaf greenness

and the overall nutrient status of the plant. When single elements are withheld from an otherwise

complete fertilizer, deficiency symptoms in angelonia for N, P, and K all appear at the same

number of days after start of treatment (Williams, 2004). An indication of lower N content in the

leaves, therefore, can help identify the overall nutrient status of the plant.

The fertilizer x cultivar interaction was highly significant at 3 and 6 weeks for both the

upper and lower leaves (Table 4-1). The trend in all cultivars, in the upper and lower leaves, at

both sampling intervals was an increase in the SPAD value as the fertilizer concentration

increased. However, the magnitude of the change varied among cultivars. At 3 weeks, SPAD

values in the lower leaves ranged from 36 to 41 at 12.5 mg-L-1 and 52 to 59 at 400 mg-L1.

Values in the upper leaves ranged from 41 to 51 and 54 to 59 at the lowest and highest

concentrations, respectively (data not shown). Since the trends observed at 3 weeks are similar

to those observed at 6 weeks, this discussion will be limited to the 6-week data.

At 6 weeks, differences between cultivars were more pronounced. SPAD values in the

lower leaves ranged from 15 to 26 at the lowest concentration and 47 to 59 at the highest

concentration (Fig. 4-3). Values in the upper leaves ranged from 31 to 43 and 54 to 59 at the









lowest and highest concentrations, respectively (Fig. 4-4). In the lower leaves, 'Angelface

White' and 'AngelMist Purple Stripe' had the lowest and highest SPAD values at 12.5 mg-L-1,

and 'Serena White' and 'Angelface White' had the lowest and highest values at 400 mg-L1.

'Serena White' was the only cultivar with a linear response, with all others quadratic (Table 4-3).

In the upper leaves, 'AngelMist Lavender Stripe' and 'Angelface White' had the lowest and

highest values at 12.5 mg-L-1, and 'AngelMist Dark Lavender' and 'AngelMist Lavender Stripe'

had the lowest and highest values at 400 mg-L-1. 'Angelface White' had the lowest SPAD value

at 12.5 mg-L-1 in the lower leaves, but also the highest value at 400 mg-L-1 in the upper leaves,

which may be an indication of increased nitrogen partitioning or mobility in this cultivar relative

to the others.

There was little difference in the 6 week SPAD values for lower and upper leaves of

plants in the 200 and 400 mg-L-1 treatments. At 400 mg-L-1, the ranges for the lower and upper

leaves were 50 to 59 and 54 to 59, respectively. At 200 mg-L-1, the ranges for the lower and

upper leaves were 38 to 51 and 50 to 56, respectively. At 100 mg-L-1 and below, the ranges no

longer overlapped. The ranges for 100 mg-L-1 were 32 to 42 for the lower leaves and 47 to 52

for the upper leaves. Even though the lower and upper leaf values no longer overlapped, the

plants did not visually look chlorotic. At concentrations below 100 mg-L-1, however, all

cultivars at all concentrations were visually chlorotic (Fig. 4-5). SPAD values in the upper

leaves at 12.5 mg-L-1, the lowest concentration used, were comparable to the lower leaf values

recorded at 100 mg-L-1. The lower leaves of the plants subjected to the 12.5 and 25 mg-L-1

treatments exhibited symptoms of nutrient deficiencies, including purpling of the leaves, light

green or yellow coloration, and marginal necrosis. As the fertilizer concentration became a









limiting factor to plant growth, mobile nutrients were mobilized from the lower leaves to the

upper leaves and meristems in order to support continued growth.

Plant Growth

The interaction was significant for height, width, size, and dry weight, indicating that the

cultivars responded differently to changes in fertilizer levels. Trends for plant height and width

are similar to those for plant size, so only the data for plant size will be presented.

Plant size increased as the fertilizer concentration increased up to maximum value, then

decreased slightly (Fig. 4-6). Regression equations were significant for all cultivars except

'Angelface Blue' and quadratic in nature (Table 4-4). Maximum plant size was obtained

between 180 and 230 mg-L1.

'Serena White' and 'Angelface White' were the smallest plants over the range of fertilizer

concentrations and 'AngelMist Lavender Stripe' and 'AngelMist Purple Stripe' were the largest.

Both the two smallest and two largest cultivars had very different growth habits which inversely

contributed to the determination of plant size. Of the two smallest cultivars, 'Serena White' is

very short and compact in habit, while 'Angelface White is naturally tall and narrow with very

little lateral branching. Of the two largest cultivars, 'AngelMist Lavender Stripe' is the shorter

cultivar but has more of a spreading habit than 'Serena White', while 'AngelMist Purple Stripe'

is the tallest cultivar in this experiment and develops lateral branches shortly after the start of

flowering (for additional descriptions of cultivars, see Chapter 2).

Changes in plant size were due to changes in both plant height and width. As fertilizer

concentrations increase from suboptimal to optimal, a small increase in concentration will have a

large positive effect on plant growth. As fertilizer concentrations continue to increase beyond

what is required for normal plant growth, elevated soluble salts levels tend to occur in the

substrate, especially in subirrigated containers where leaching does not occur. This increase, as









monitored by EC levels, can result in decreased plant growth (Whipker et al., 2003). The

optimal fertilizer concentration for plant size in this experiment was around 200 mg-L-1 for most

cultivars.

The interaction for plant dry weight was significant. For those cultivars that had a

quadratic response to increased fertilization, dry weight (g) per plant increased as the

concentration applied increased up to a maximum between 232 and 272 g, and then decreased at

higher concentrations (Fig. 4-7). In 'AngelMist Lavender Stripe', the response was linear and

did not reach a plateau at the concentrations tested. 'Angelface Blue' did not have a significant

regression equation, and the highest observed value was at 100 mg-L-1. In general, cultivars that

were larger in plant size tended to have higher dry weights. 'AngelMist Lavender Stripe' and

'AngelMist Purple Stripe' had the highest dry weights over all concentrations except at 12.5

mg-L-1, and 'AngelMist Dark Lavender', 'Angelface White', and 'Serena White' had the lowest.

As the fertilizer concentration increased from a deficient level to an optimal level, the plant dry

weight increased. Above an optimal level, the high EC levels in the leachate inhibited growth to

some degree and plants accumulated less dry weight on a per plant basis.

Flowering

The cultivar x fertilizer interaction for number of inflorescences per plant was significant.

Large differences were observed in the responses of the cultivars to increased fertilization. The

general trend was an increase in the number of inflorescences as fertilizer concentration

increased. Between 200 and 400 mg-L-1' inflorescence number either declined slightly or

remained unchanged (Fig. 4-8). 'Angelface White' and 'Serena White' had the least and greatest

number of inflorescences, respectively, at all concentrations. The three cultivars with the lowest

number of inflorescences 'Angelface White', 'Angelface Blue', and 'AngelMist Purple Stripe'

- are cultivars that tend to have fewer inflorescences per plant and a greater number of flowers









per inflorescence (see Chapter 2). The interaction for number of days to first flower was not

significant. The differences observed were due to the main effects of cultivar, but not fertilizer

concentration (Table 4-5).

The average number of days to first flower in this experiment was very similar to the

results of a cultivar screen conducted one month earlier in the same greenhouse with these

cultivars (see Chapter 2). 'Serena White' flowered in an average of 22 d and was the earliest to

flower relative to all other cultivars except for 'AngelMist Dark Lavender'. 'Angelface White'

was the latest to flower and averaged 43 d. It was almost 2 weeks later than 'AngelMist Purple

Stripe' and nearly 3 weeks later than the other cultivars. Flowering was not significantly

enhanced or delayed by fertilizer concentration. These results are in contrast to Salvia splendens,

in which flowering was delayed by 8 d at lower fertilizer concentrations (Kang and van lersal,

2004). Thus, the length of the crop cycle in angelonia will be unaffected if slight nutritional

deficiencies are induced as a means to control crop vegetative growth during production.

Conclusions

There was not a particular fertilizer concentration that yielded maximum growth and

flowering for all parameters. The response of cultivars to increased fertilizer concentration was

not cultivar-specific, indicating that they will respond similarly to changes in the concentration

applied. Leaf greenness, a primary visual indicator of the nutritional status of a plant, increased

in both the upper and lower leaves as the fertilizer concentration applied increased. Maximum

plant size occurred between 180 and 230 mg-L-1, maximum plant dry weight (g) occurred

between 230 and 275 mg-L-1, and maximum inflorescence number occurred between 245 and

340 mg-L-1. The recommended EC range (2.0 to 3.2 mS-cm-1) fell between the 100 and 200

mg-L-1 treatments.









The selection of a fertilizer concentration is based upon many factors, including the

concentration required for acceptable plant quality, the irrigation method used, and the cost of

the fertilizer. Best management practices recommend using the least amount of fertilizer

necessary to ensure adequate growth and development of the crop. Although maximum plant

growth occurred at concentrations greater than 100 mg-L-1, it was the lowest fertilizer

concentration at which all cultivars were not visually chlorotic. Plant growth and quality were

acceptable for the retail market. Recommendations from this experiment would be to use a

fertilizer concentration of 100 mg-L-1 or slightly higher for the warm-season production of

subirrigated angelonia, regardless of cultivar. Slight adjustments may need to be made for

climate and time of year.

Previous recommendations for angelonia fertilization ranged from N at 75 to 200 mg-L1.

Differences in recommendations were not defined as being specific to any cultivar or irrigation

method. Typically, fertilizer recommendations are based upon overhead irrigation. Since no

leaching occurs in subirrigation systems, it is generally recommended that growers use a half-

strength fertilizer solution when using subirrigation instead of overhead irrigation (Nelson,

1994). The results from this study agree with production guidelines of overhead irrigating with

150 to 200 mg-L-1 as published for these cultivars by their suppliers and by Schoellhorn and

Alvarez (2002). However, this range is much narrower than what had been published in the

trade press.















6.5


6.0


5 5.5


5.0


4.5


4.0
0 50 100 150 200 250 300 350 400
Fertilizer N concentration (mg*L"1)

Figure 4-1. Effect of increasing fertilization with 20.0N-4.4P-16.6K on substrate pH in
Angelonia angustifolia 6 weeks after start of treatments. Treatment N concentrations
were 12.5, 25, 50, 100, 200, or 400 mg-L1. The regression line was generated from
individual plant data and the points on the graph are the means for each concentration
pooled over cultivar (n=24). The equation and r2 value can be found in Table 4-2.


Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White
Ser White AM Dk Lav A AM Lav Stripe I AM Purp Stripe AF White


- AF Blue
A AF Blue


0 50 100 150 200 250 300 350 400
Fertilizer N concentration (mg*L"1)


Figure 4-2. Effect of increasing fertilization with 20.0N-4.4P-16.6K on substrate electrical
conductivity in Angelonia angustifolia 6 weeks after start of treatments. Treatment N
concentrations were 12.5, 25, 50, 100, 200, or 400 mg-L-1. The regression lines were
generated from individual plant data and the points on the graph are the means for
each individual treatment (n=8). Regression equations and r2 values can be found in
Table 4-2.











Ser White AM Dk Lav -AM La Stripe -AM Purp Stripe AF White --AF Blue
Ser White AM Dk La A AM Lav Stripe AM Purp Stripe AF White A AF Blue


60


50


Q 40-
a-

30


20


10
0 50 100 150 200 250 300 350 400
Fertilizer N concentration (mgL"1)

Figure 4-3. Effect of increasing fertilization with 20.0N-4.4P-16.6K on lower leaf SPAD values
in Angelonia angustifolia 6 weeks after start of treatments. Treatment N
concentrations were 12.5, 25, 50, 100, 200, or 400 mg-L1. SPAD units indicate
relative leaf greenness. The regression lines were generated from individual plant
data and the points on the graph are the means for each individual treatment (n=8).
Regression equations and r2 values can be found in Table 4-3.



Ser White AM Dk Lav -AM LavStripe -AM PurpStripe AF White -AF Blue
Ser White AM Dk Lav A AM Lav Stripe U AM Purp Stripe AF White A AF Blue

60

55

50




40

35

30
0 50 100 150 200 250 300 350 400
Fertilizer N concentration (mgL"1)

Figure 4-4. Effect of increasing fertilization with 20.0N-4.4P-16.6K on upper leaf SPAD values
in Angelonia angustifolia 6 weeks after start of treatments. Treatment N
concentrations were 12.5, 25, 50, 100, 200, or 400 mg-L-1. SPAD units indicate
relative leaf greenness. The regression lines were generated from individual plant
data and the points on the graph are the means for each individual treatment (n=8).
Regression equations and r2 values can be found in Table 4-3.























II A I 8 I B IC l 1













D E F


Figure 4-5. Response ofAngelonia angustifolia 'AngelMist Dark Lavender' to increasing rates
of fertilization with 20.0N-4.4P-16.6K. N concentrations listed are in mg-L1.
A) 12.5 B) 25 C) 50 D) 100 E) 200 F) 400.











Ser White AM Dk Lav -AM Lav Stripe -AM Purp Stripe AF White AF Blue
Ser White AM Dk Lav A AM Lav Stripe U AM Purp Stripe AF White A AF Blue


70


60-
E A

50-


40


30
0 50 100 150 200 250 300 350 400
Fertilizer N concentration (mg.L"1)

Figure 4-6. Effect of increasing fertilization with 20.0N-4.4P-16.6K on plant size inAngelonia
angustifolia 6 weeks after start of treatments. Treatment N concentrations were 12.5,
25, 50, 100, 200, or 400 mg-L-1. Plant size calculated as [(plant height + average
plant width)/2]. The regression lines were generated from individual plant data and
the points on the graph are the means for each individual treatment (n=8). Regression
equations and r2 values can be found in Table 4-4.



Ser White AM Dk Lav -AM Lav Stripe -AM Purp Stripe AF White AF Blue
Ser White AM Dk Lav A AM Lav Stripe U AM Purp Stripe AF White A AF Blue

70 -

60

S50

S40

30 /

20 /-

10 -

0
0 50 100 150 200 250 300 350 400
Fertilizer N concentration (mg.L"1)

Figure 4-7. Effect of increasing fertilization with 20.0N-4.4P-16.6K on plant dry weight in
Angelonia angustifolia 6 weeks after start of treatments. Treatment N concentrations
were 12.5, 25, 50, 100, 200, or 400 mg-L-1. The regression lines were generated from
individual plant data and the points on the graph are the means for each individual
treatment (n=8). Regression equations and r2 values can be found in Table 4-4.











Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue
Ser White AM Dk Lav A AM Lav Stripe I AM Purp Stripe AF White A AF Blue


A


0 50 100 150 200 250 300 350 400
Fertilizer N concentration (mg.L"1)


Figure 4-8. Effect of increasing fertilization with 20.0N-4.4P-16.6K on number of
inflorescences in Angelonia angustifolia 6 weeks after start of treatments. Treatment
N concentrations were 12.5, 25, 50, 100, 200, or 400 mg-L-1. The regression lines
were generated from individual plant data and the points on the graph are the means
for each individual treatment (n=8). Regression equations and r2 values can be found
in Table 4-4.










Table 4-1. Analysis
3 wk
SPAD
upper
leaves
1 ***


of variance for Angelonia angustifolia cultivars in response to fertilization
3 wk 6 wk 6 wk 3 wk 6 wk 3 wk 6 wk Plant Plant
SPAD SPAD SPAD pH pH EC EC height width
lower upper lower (cm) (cm)
leaves leaves leaves
*** *** *** *** *** *** *** *** ***


Plant
size
(cm)


Dry
weight
(g)


Inflor. Days
Number to
flower


I 1 L111LI1
Cultivar *** *** ** *** ** NS NS *** *** *** *
Fert x Cv *** *** *** *** *** NS NS *** *** *
Ns, **, *** nonsignificant and significant at P<0.05, 0.01, and 0.001, respectively.

Table 4-2. Regression equations for pH and EC measurements collected at 6 weeks after treatment.
Variable Cultivarz Significancey r2 Equation
pH Q*** 0.73 y = 6.4 0.01x + 0.00002x2


EC (mS-cm-1) SW Q* 0.76 y = 0.1 + 0.02x 0.00003x2
AMDL Q* 0.66 y = 0.1 + 0.03x 0.00004x2
AMLS Q* 0.94 y = 0.6 + 0.001x + 0.00001x2
AMPS L*** 0.93 y = 0.5 + 0.001x
AFW L*** 0.86 y = 0.6 + 0.01x
AFB Q* 0.81 y = 0.3 + 0.02x 0.00003x2
z Cultivar abbreviations: SW 'Serena White', AMDL 'AngelMist Dark Lavender', AMLS 'AngelMist Lavender Stripe',
AMPS 'AngelMist Purple Stripe', AFW 'Angelface White', AFB 'Angelface Blue'.
Y *, *** significant at P<0.05, 0.01, and 0.001, respectively.









Table 4-3. Regression equations for SPAD readings collected at 6 weeks after treatments.
Variable Cultivarz Significancey r2 Equation
6 week SPAD SW Q** 0.58 y = 41.7 + 0.08x 0.0001x2
(upper leaves) AMDL Q* 0.37 y = 45.3 + 0.07x 0.0001x2
AMLS Q*** 0.66 y = 35.7 + 0.15x 0.0002x2
AMPS Q*** 0.57 y = 39.9 + 0.14x 0.0003x2
AFW Q* 0.54 y = 44.8 + 0.07x 0.0001x2
AFB Q* 0.67 y = 39.0 + 0.08x 0.0001x2

6 week SPAD SW Q*** 0.76 y = 22.3 + 0.16x 0.0002x2
(lower leaves) AMDL L*** 0.32 y = 47.4 + 0.02x
AMLS Q*** 0.83 y = 20.6 + 0.23x 0.0004x2
AMPS Q*** 0.85 y = 23.2 + 0.19x 0.0003x2
AFW Q*** 0.92 y = 13.1 + 0.20x 0.0002x2
AFB Q*** 0.87 y = 25.4 + 0.16x 0.0002x2
SCultivar abbreviations: SW 'Serena White', AMDL 'AngelMist Dark Lavender', AMLS 'AngelMist Lavender Stripe',
\o AMPS 'AngelMist Purple Stripe', AFW 'Angelface White', AFB 'Angelface Blue'.
Y *, **, *** significant at P<0.05, 0.01, and 0.001, respectively.









Table 4-4. Regression equations for plant size, dry weight, and number of inflorescences at 6 weeks after treatment.
Variable Cultivarz Significancey r2 Equation


Plant size (cm)


Dry weight (g)







Inflorescence Number


SW
AMDL
AMLS
AMPS
AFW
AFB

SW
AMDL
AMLS
AMPS
AFW
AFB

SW
AMDL
AMLS
AMPS
AFW
AFB


Q***
Q***
Q*
Q***
Q*
NS

Q*
Q*
L*
Q***
Q***
NS

Q***
L**
Q***
Q*
L***
Q*


41.5 + 0.09x
47.3 +0.09x-
59.5 + 0.04x
55.0 + 0.14x
47.4 + 0.05x


13.0 + 0.15x
15.4 + 0.12x
32.2 + 0.08x
9.8 + 0.50x -
18.4 + 0.16x


- 0.0002x2
- 0.0002x2
- O.0001x2
- 0.0003x2
- O.0001x2


- 0.0003x2
- 0.0002x2

0.001x2
- 0.0003x2


0.33
0.36
0.14
0.36
0.14


0.43
0.27
0.26
0.52
0.58


0.67
0.20
0.58
0.20
0.41
0.46


z Cultivar abbreviations: SW 'Serena White', AMDL 'AngelMist Dark Lavender', AMLS -
AMPS 'AngelMist Purple Stripe', AFW 'Angelface White', AFB 'Angelface Blue'.
y NS, *** nonsignificant and significant at P<0.05, 0.01, and 0.001, respectively.


'AngelMist Lavender Stripe',


23.4 + 0.57x 0.001x2
35.4 + 0.07x
20.3 + 0.19x- 0.0003x2
4.0 + 0.04x 0.00008x2
0.6 + 0.006x
4.6 + 0.05x 0.00007x2









Table 4-5. Number of days to first open flower in Angelonia angustifolia. The fertilizer x
cultivar interaction and the main effect of cultivar were not significant at P<0.05. The
main effect of cultivar displayed in this table are pooled over all fertilizer
concentrations for each cultivar. Mean separation was performed using Tuley's at
a=0.05.
Cultivarz Number of Mean
days separation
AFW 43.0 a
AMPS 29.9 b
AMLS 26.9 c
AFB 25.5 cd
AMDL 23.8 de
SW 22.2 e
z Cultivar abbreviations: SW 'Serena White', AMDL 'AngelMist Dark Lavender', AMLS -
'AngelMist Lavender Stripe', AMPS 'AngelMist Purple Stripe', AFW 'Angelface White',
AFB 'Angelface Blue'









CHAPTER 5
Angelonia angustifolia CULTIVARS DIFFER IN RESPONSE TO PACLOBUTRAZOL,
DAMINOZIDE, AND ETHEPHON

Introduction

Commercially-available Angelonia angustifolia cultivars differ significantly in growth

habit (Chapter 2), and many tend to become excessive in height during production without the

use of plant growth regulators (PGRs). Commonly used PGRs on bedding plants include

chlormequat chloride, daminozide, ethephon, paclobutrazol, and uniconazole. With the

exception of ethephon, all inhibit cell elongation by blocking gibberellin synthesis (Gent and

McAvoy, 2002). Ethephon releases ethylene within plant tissues, which can result in shorter

internodes and increased lateral branching (Beaudry and Kays, 1988). Plant response to PGR

applications varies by chemical and concentration applied (Barrett, 2001), method of application

(Gibson and Whipker, 2003), and species or cultivar sensitivity (Gibson and Whipker, 2001a;

Lewis et al., 2004; Starman et al., 2004; Warner and Erwin, 2003). Cultivar differences in

response to PGR applications are often due to differences in sensitivity to the PGR applied, and

this may or may not correlate with plant vigor (Ecke III et al., 2004).

Research indicates that at least some angelonia cultivars are responsive to daminozide,

ethephon, and paclobutrazol applications. 'Blue Pacific' plants treated with 5000 mg-L-1

daminozide were shorter than untreated plants, but paclobutrazol sprays of up to 80 mg-L-1 were

ineffective at controlling height (Starman, 2001). Miller and Armitage (2002), however,

observed that five cultivars in the AngelMist series had similar responses to PGRs and that

paclobutrazol sprays at 50 and 100 mg-L-1, as well as ancymidol at 50 and 100 mg-L-1 and

daminozide at 2500 and 5000 mg-L-1 effectively controlled plant height. It is unknown if current

cultivars and/or series will vary in their sensitivity to the type and concentration of PGR applied.









Published recommendations for PGR applications for growth control of angelonia suggest

spraying daminozide at 2500 mg-L-1, uniconazole at 2 to 5 mg-L-1; paclobutrazol at 5 mg-L-1; or a

tank mix of chlormequat chloride plus daminozide at 700 to 1000 and 1500 to 2000 mg-L-1,

respectively (Ball FloraPlant, 2006; PanAmerican Seed, 2006; Proven Winners, 2006; Selecta,

2008). Proven Winners recommends avoiding the use of ethephon on cultivars in the Angelface

series due to its ability to cause leaf tip burn or distorted growth, whereas the other companies

neither recommend using or avoiding this chemical. It is unknown whether the phytotoxicity

symptoms observed in the Angelface series are series-specific.

The objectives of these experiments were 1) to determine which PGR chemicals and which

concentrations will effectively control growth of angelonia, and 2) whether or not response to the

PGRs is cultivar specific or not. Four experiments were conducted: a rate screen examining the

efficacy of daminozide, ethephon, and paclobutrazol on seven cultivars, and three more in-depth

studies, each focusing solely on daminozide, paclobutrazol, or ethephon in order to look at

differences between representative cultivars under different growing conditions.

Materials and Methods

Experiment 5-1 Paclobutrazol, Daminozide, and Ethephon Response Curves

Cuttings from seven Angelonia angustifolia cultivars 'Angelina Violet and White',

'AngelMist Dark Lavender', 'AngelMist Purple Improved', 'AngelMist Lavender Stripe',

'AngelMist Purple Stripe', 'Angelface White', and 'Angelface Blue' were harvested from

stock plants on 14 May 2007 (see Expt. 2-2 for stock plant culture). Sixty rooted cuttings of

each cultivar were planted on 12 June, into 2.2 L (16 cm diameter) pots filled with Fafard 2

soilless medium (Conrad Fafard, Inc., Apopka, FL) and pinched to two nodes. Two weeks later,

on 26 June, the following plant growth regulator (PGR) treatments were applied: paclobutrazol

(Bonzi; Syngenta Corp., Greensboro, NC) drenches at 5, 10, or 20 mg-L-1; ethephon (Florel;









Monterey, Fresno, CA) sprays at 250, 500, or 1000 mg-L-1 plus Capsil (Aquatrols Corp. of

America; Paulsboro, NJ) at 0.5mL-L-1; daminozide (B-Nine; OHP, Mainland, PA) sprays at

1250, 2500, or 5000 mg-L-1; and an untreated control. Drench volume was 180 ml per pot for

paclobutrazol, and spray volumes were 306 mL-m-2 for ethephon and 204 mL-m-2 for

daminozide, respectively.. Plants were grown in a naturally ventilated greenhouse and

continuously liquid-fed as needed with N at 150 mg-L-1 using 20.0N-4.4P-16.6K (Peters 20-10-

20 Florida Special, The Scotts Co., Marysville, OH). Average daily minimum, maximum, and

24-hour temperatures over the length of the experiment were 22.7, 35.9, and 27.9 C, and the

average daily light integral (DLI) was 17.8 molm-2-d-1.

Plant height, measured from the rim of the pot to the tallest point on the plant, was

measured at treatment and again 23 d later, on 19 July, once plants reached a marketable stage.

Stem elongation was calculated as the difference between the two heights. The experiment with

10 PGR treatments and seven cultivars was set up in a split plot design, with PGR as the main

plot and cultivar as the sub-plot. Plants were randomized in a complete random block design

with three blocks and two plants of each cultivar per block. Data were analyzed by regression

analysis in SAS 9.1 (SAS Institute, Cary, NC) in order to generate response curves for each

cultivar. Each PGR was analyzed individually, with a common untreated control used for each

cultivar for all three growth regulators.

Experiment 5-2 Daminozide

The cultivars 'AngelMist Dark Lavender', 'AngelMist Purple Stripe', and 'Angelface

White' were selected from those used in Expt. 5-1 because they were examples of a wide range

of growth habits. See Expt. 5-1 for description of materials and methods. Forty-eight rooted

cuttings of each cultivar were transplanted into 2.2 L (16 cm diameter) pots on 11 July 2007.

Daminozide (B-nine; OHP, Mainland, PA) sprays were applied on 27 July, at 0, 1250, 2500, or









-2
5000 mg-L-1 at a spray volume of 204 mL-m 2. Average daily minimum, maximum, and 24-hour

temperatures over the length of the experiment were 23.8, 37.5, and 29.1 C, and average DLI

was 18.8 mol-m-2day-1. Final height was measured on 22 Aug. and stem elongation was

calculated. The experiment was set up as a factorial with four daminozide concentrations and

three cultivars. Plants were randomized in a complete random block design with three blocks

and four plants of each treatment per block. Analysis of variance and Tukey's mean separation

at a=0.05 were conducted using SAS.

Experiment 5-3 Paclobutrazol Spray and Drench Applications

See Expt. 5-1 for materials and methods. Unrooted cuttings from 'AngelMist Purple

Stripe' and 'Angelface White' were harvested on 25 July 2007, and 81 rooted cuttings of each

cultivar were transplanted into 2.2 L (16 cm diameter) pots on 17 Aug. Paclobutrazol sprays

-2
were applied 2 weeks after pinch at 50 or 100 mg-L-1 at a volume of 204 mL-m 2. For drench

applications, paclobutrazol concentrations were 2, 4, or 8 mg-L-1 using a drench volume of 180

mL per pot. Applications were made at visible bud, which was 4 and 6.5 weeks after pinch for

'AngelMist Purple Stripe' and 'Angelface White', respectively. Average daily minimum,

maximum, and 24-hour temperatures over the length of the experiment were 22.7, 34.5, and 27.1

C, and average DLI was 14.8 mol-m-2day-1. Final height was measured when control plants

reached a marketable stage, on 28 Sept. and 15 Oct. for 'AngelMist Purple Stripe' and

'Angelface White', respectively, and stem elongation was calculated. Plants were arranged in a

randomized complete block design with 3 blocks and 3 plants of each treatment per block.

Analysis of variance and mean separation using Tukey's a=0.05 were conducted using SAS.

Experiment 5-4 Ethephon

Rooted liners of 'AngelMist Dark Lavender', 'AngelMist Purple Improved', and

'AngelMist Purple Stripe' from Ball FloraPlant (West Chicago, IL) and rooted liners of









'Angelface White Improved' from EuroAmerican Propagators (Bonsall, CA) were received the

week of 1 Oct. 2007. Forty-eight plants of each cultivar were planted on 8 Oct. into 650 cm3 (11

cm) pots filled with Fafard 2-P soilless medium (Conrad Fafard, Inc.; Apopka, FL) and pinched

to two nodes. Ethephon applications were applied 10 d after transplant, on 18 Oct., at

concentrations of 0, 250, 500, or 1000 mg-L-1 using techniques given in Expt. 5-1. This

experiment was designed as a factorial consisting of four cultivars and four ethephon

concentrations. Plants were arranged in a randomized complete block design, with three blocks,

16 treatments, and three plants per treatment per block. They were grown on a subirrigation

bench and fertilized with N at 75 mg-L-1 using 20.0N-4.4P-16.6K (Peters 20-10-20 Florida

Special, The Scotts Co., Marysville, OH). Average daily maximum, minimum, and 24-hour

temperatures and DLI during the first week of the experiment were 31.5, 19.5, and 24.6 C and

9.8 mol-m-2day-1, and during the last week were 21.2, 13.4, and 16.7 C and 7.8 mol-m-2day-1,

respectively.

At 1 week after treatment (1 WAT), plants were visually rated for appearance of

phytotoxicity symptoms on a 0 3 scale, with 0 = no visible symptoms, 1 = slight chlorosis of

leaf tips, 2 = moderate chlorosis of leaves and slight necrosis at leaf edges, and 3 = severe

chlorosis of leaves, moderate to severe leaf necrosis, and leaf distortion. Plant height was

measured at 4 WAT and stem elongation was calculated. The number of laterals per plant was

determined at 5 WAT. All shoots greater than 5 cm were considered as laterals, and no

distinction was made as to whether they were primary or secondary laterals. Plants were

observed daily and date of first flower was recorded.

Analysis of variance was determined for all parameters using SAS. Regression analysis

was run by cultivar on plant height, width, and size, and number of days to flower. Mean









separation using Tukey's at a=0.05 was determined for phytotoxicity rating and number of

laterals per plant.

Results and Discussion

Experiment 5-1

The chemical x cultivar interaction was significant. Daminozide, ethephon, and

paclobutrazol were effective at controlling plant growth, but their effectiveness varied with the

cultivar and concentration applied. For each PGR, the concentrations selected were based upon

published recommendations. The middle concentration was the one recommended for

production, and the higher and lower concentrations were selected in order to generate response

curves.

Paclobutrazol

All cultivars exhibited a significant quadratic response to paclobutrazol, with a general

decrease in stem elongation as the concentration increased (Table 5-1, Fig. 5-la). All of the

regression curves showed a strong linear component between the untreated control plants and the

5 mg-L-1 concentration, and as the concentration applied increased from 5 to 20 mg-L-1, there

was a smaller incremental reduction in stem elongation.

Cultivars varied in their response to paclobutrazol. At 5 mg-L-1, stem elongation in

treated plants was 22 to 65 percent of untreated controls. 'Angelina Violet and White' was the

most sensitive cultivar and 'AngelMist Purple Improved' and 'AngelMist Dark Lavender' were

the least sensitive cultivars. The percent stem elongation observed in 'Angelina Violet and

White' at 5 mg-L-1 was comparable to 'Angelface Blue' or 'AngelMist Purple Improved' plants

treated with 20 mg-L-1 (data not shown).

Paclobutrazol drenches at 5 mg-L-1 have been recommended for height control of

angelonia cultivars grown in warm climates (PanAmerican Seed, 2006). However, 5 mg-L-1 was









too high for most cultivars, even though applications were applied during the summer. In

'AngelMist Purple Stripe,' a vigorous cultivar, the efficacy of the application had started to

decrease by the time data was collected and the intemodes in the inflorescence were beginning to

elongate almost normally (Figure 5-2). Concentrations less than 5 mg-L-1 should be trialed on an

individual cultivar basis for a more precise determination of optimal drench concentration.

Daminozide

Stem elongation decreased as the concentration applied increased in all cultivars (Fig. 5-

lb, Table 5-1). 'Angelina Violet and White' and 'AngelMist Purple Stripe' had significant

quadratic responses and the other five cultivars exhibited linear responses to increasing

daminozide concentration. Stem elongation in plants sprayed with 5000 mg-L-1 was 62 to 87

percent of the untreated controls. 'Angelina Violet and White' and 'Angelface Blue' were the

two most responsive cultivars. 'AngelMist Dark Lavender' was the least responsive cultivar.

'AngelMist Purple Stripe' and 'Angelface Blue' had the greatest stem elongation at all

concentrations applied. Both of these cultivars are tall plants and naturally have longer

internodes than more compact cultivars. At concentrations of 2500 mg-L-1 and below, both

cultivars had similar responses to daminozide. However, at 5000 mg-L-1, 'Angelface Blue' had a

greater response to the application than 'AngelMist Purple Stripe'(Fig. 5-1b).

The recommended spray concentration for daminozide is 2500 mg-L-1 (Selecta, 2008), and

it is applied at much higher concentrations than paclobutrazol because it has much lower activity

(Barrett, 2001). It is commonly used on bedding crops in a tank mix solution with chlormequat

chloride. The concentration of daminozide used varies depending upon the concentration of

chlormequat chloride used, but ranges from 1500 to 2500 mg-L-1. Daminozide applied by itself

is not as effective as using a tank mix, but can be effective in certain crops.









The cultivars in this experiment varied in their response to daminozide, from moderately

effective to ineffective (Fig. 5-3). This is similar to the varied response observed in a trial of 26

ornamental cabbage and kale (Brassica oleracea) cultivars in which efficacy was observed in

two cultivars when sprayed with 2500 mg-L-1 and in eight cultivars when sprayed with 5000

mg-L-1 (Gibson and Whipker, 2001a).

Ethephon

The general trend observed with ethephon was that stem elongation decreased as the

concentration applied increased (Fig. 5-2c, Table 5-1). All of the cultivars had significant linear

regression responses. 'AngelMist Purple Stripe' was the most responsive cultivar and

'AngelMist Dark Lavender' was the least responsive cultivar. Stem elongation in plants treated

with 1000 mg-L-1 ranged from 54 to 80 percent of untreated plants, respectively.

The efficacy of ethephon in controlling height of vegetative annuals is both crop and

cultivar specific. Hammond et al. (2007) observed that blanketflower (Gaillardiapulchella)

'Torch Flame' was more compact than untreated plants but a native Florida ecotype was

insensitive to ethephon applications. Starman et al. (2004) observed in a wide selection of

vegetative annuals that 81% were responsive to ethephon applications. Multiple cultivars were

trialed within seven species and cultivar differences were observed in Antirrhinum majus,

Calibrachoa hybrids, and Petunia xhybrida, but not in Diascia xhybrida, Impatiens wallerana,

Lantana camera, or Nemesia xhybrida.

Experiment 5-2

The cultivar x daminozide concentration interaction was not significant, but the main

effects of cultivar and concentration were significant. Within each concentration, the averages

were pooled across cultivars, and within each cultivar, the averages were pooled across

concentrations for mean separation (Table 5-2).









For the main effects of concentration, all concentrations were significantly different from

the untreated control and a rate response was observed. Stem elongation was not significantly

different between 1250 and 2500 mg-L-1, but 5000 mg-L-1 was significantly different from the

other treatments. These results are similar to those observed in Expt. 5-1, in which an increase in

concentration decreased stem elongation.

All three cultivars had similar responses to daminozide, even though they had different

growth habits. In Expt. 5-1, stem elongation in plants sprayed with 5000 mg-L-1 was 73 to 82

percent of untreated plants, depending upon cultivar, and in Expt. 5-2, stem elongation was 78 to

87 percent of untreated plants. Plants were slightly less responsive in Expt. 5-2 due to the higher

average daily temperatures under which the plants were grown. 'Angelface White' was the most

responsive cultivar to daminozide in both experiments. 'AngelMist Dark Lavender' had the

greatest decrease in sensitivity between the two experiments, in which plants treated with 5000

mg-L-1 were 75 percent the height of untreated plants in Expt. 5-1 but 87 percent in Expt. 5-2.

For the main effects of cultivar, all three cultivars were significantly different from each

other. These differences can be explained based upon each cultivar's natural growth habit (see

Chapter 2). Plant height, from tallest to shortest, was 'AngelMist Purple Stripe', 'Angelface

White', and 'AngelMist Dark Lavender', respectively. 'AngelMist Purple Stripe' and

'Angelface White' are both tall, vigorous cultivars and 'AngelMist Dark Lavender' is a short,

compact cultivar.

Daminozide applications were effective at controlling plant height, but a high

concentration is needed for efficacy. In warm climates and during the summer months, it is not

the most effective PGR for this crop. However, in the cooler months and in northern states, it









may be a viable option since angelonia cultivars are more responsive to daminozide than

paclobutrazol sprays.

Experiment 5-3

The cultivar x treatment interaction (with treatment defined as the drench and spray

applications at varying concentrations) and the main effect of paclobutrazol treatment, were not

significant, but the effect of cultivar was significant (Table 5-3). 'AngelMist Purple Stripe' is a

slightly taller cultivar than 'Angelface White' at marketability in the late summer. None of the

treatments resulted in plants with less stem elongation than untreated plants.

The paclobutrazol spray applied 2 weeks after pinch was not effective at controlling plant

height, which is consistent with results from Starman (2001) but inconsistent with results

observed by Miller and Armitage (2002). The later drenches applied at visible bud in this

experiment were not effective at controlling plant height either, but most likely because they

were applied too late in production to significantly alter final height. Both of these cultivars are

tall and have vigorous growth habits and will need to receive a paclobutrazol drench, rather than

a spray, within the first 2 to 3 weeks after pinch in order to obtain adequate height control.

'AngelMist Purple Stripe' was treated 4 weeks after pinch, and by then it had already grown too

tall.

Paclobutrazol drenches at 5 mg-L-1 have been recommended for height control of

angelonia cultivars grown in warm climates (PanAmerican Seed, 2006). Uniconazole, another

triazole PGR, has been recommended as a drench application at 2 to 5 mg-L-1 (Proven Winners,

2006). Uniconazole is more active than paclobutrazol and in general, paclobutrazol

concentrations are required to be two to four times higher than uniconazole in order to get the

same degree of efficacy (Barrett, 2001).









Based on these recommendations, drench applications of 5 to 10 mg-L-1 paclobutrazol

should have resulted in marketable plants but instead resulted in severely stunted plants. Early

sprays, which have been shown to be effective at 50 and 100 mg-L-1 (Miller and Armitage,

2002), and late drenches at lower concentrations were used in Expt. 5-3 in order to examine if

the timing of the drench application affects efficacy and if early sprays would be a viable option

instead of drench application. However, these cultivars did not respond to the spray or drench

concentrations used in Expt. 5-3. Spray applications will not be practical due to the high

concentration required for efficacy. Drench applications will need to be applied carefully and

concentrations will need to be altered depending upon the age of the plant. Early in production,

lower concentrations should be effective at controlling stem elongation. Later in the crop,

drench applications will need to be applied before the presence of visible bud and at higher

concentrations than those used in the early drenches. Applications should be made before the

plant begins to elongate rapidly following transplant.

Experiment 5-4

Significant cultivar x concentration interactions occurred for phytotoxicity, stem

elongation, and number of days to flower (Table 5-4). The interaction was not significant for

number of laterals, but the main effects of cultivar and ethephon concentration were significant.

Phytotoxicity

Cultivars responded similarly to the application of ethephon at 250 mg-L-1 and very little

phytotoxicity (marginal chloroisis, marginal necrosis, and/or leaf malformation) was observed

(Table 5-5). At 500 mg-L-1, all cultivars exhibited some degree of phytotoxicity, and 'AngelMist

Dark Lavender' and 'Angelface White Improved' were more severely affected than 'AngelMist

Purple Improved' and 'AngelMist Purple Stripe.' At 1000 mg-L-1, 'AngelMist Purple Stripe'

exhibited the least phytotoxicity symptoms relative to the other three cultivars. The appearance









of phytotoxicity in all cultivars at the recommended application concentration is consistent with

cultural information published by Proven Winners (2006), the supplier of 'Angelface White

Improved.' Starman (2001), however, applied ethephon to 'Blue Pacific' and reported an

inhibition of stem elongation but did not mention the appearance of any phytotoxicity symptoms.

These phytotoxicity symptoms only appeared on the leaves that were directly sprayed with

ethephon. By 4 weeks after treatment, damaged leaves were no longer visible under the plant

canopy and the leaf morphology of the newly-expanding leaves was normal.

Stem elongation

Ethephon reduced stem elongation in all cultivars (Fig. 5-5, Table 5-6) at 4 weeks after

treatment. All cultivars except 'AngelMist Purple Improved' exhibited quadratic response

curves. The difference between the tallest and shortest cultivar at each concentration decreased

as the concentration applied increased. 'AngelMist Dark Lavender' and 'AngelMist Purple

Stripe' had very similar response curves. Stem elongation was almost identical at 4 weeks after

treatment, but most likely would not have continued if data had been taken at a later date due to

the presence of flower buds on 'AngelMist Dark Lavender', an early-flowering cultivar, at the

time of data collection.

These results are consistent with the decrease in stem elongation observed in Expt. 5-1.

Starman et al. (2004) observed that multiple cultivars of diascia, impatiens, lantana, and nemesia

all exhibited decreased stem elongation following the application of ethephon, but in snapdragon,

calibrachoa, and petunia, some cultivars responded and others did not. In this experiment,

'AngelMist Dark Lavender' and 'AngelMist Purple Stripe' had a greater response to the

ethephon applications than 'AngelMist Purple Improved' and 'Angelface White Improved.'









Lateral number

The cultivar x concentration interaction was not significant, but the main effects of cultivar

and concentration were significant (Table 5-7). 'AngelMist Dark Lavender' had a significantly

higher number of laterals compared to the other three cultivars. This may be due to the fact that

this cultivar flowers earlier than the others, and once angelonia plants begin to develop

inflorescences, the axillary meristems are released from apical inhibition and lateral branching

ensues. Since the data was collected after 'AngelMist Dark Lavender' had initiated flowering, it

had started developing secondary laterals in both treated and untreated plants.

Ethephon applications at 250 or 500 mg-L-1 were not different from the control. Ethephon

at 1000 mg-L-1 resulted in a higher number of laterals compared to 500 mg-L-1. This increase in

lateral number is consistent with results from other studies conducted on a number of bedding

plants and perennial species (Hayashi et al., 2001; Faust and Lewis, 2005).

Flowering

The application of ethephon delayed flowering in all cultivars (Fig. 5-6, Table 5-6).

'AngelMist Dark Lavender' and 'AngelMist Purple Stripe' exhibited quadratic responses. As

the ethephon concentration applied increased up to 500 mg-L-1, plants exhibited close to a linear

increase in time to flower, but between 500 and 1000 mg-L-1, the additional amount of delay

observed was relatively small. 'AngelMist Purple Improved' and 'Angelface White Improved'

exhibited linear responses. 'AngelMist Dark Lavender' was the earliest to flower at all

concentrations applied. 'AngelMist Purple Improved' and 'Angelface White' were the latest to

flower at all concentrations applied. The untreated controls in both cultivars flowered within 2 d

of each other, but as the ethephon concentration increased, 'Angelface White Improved' had

more of a delay in flowering than 'AngelMist Purple Improved.' At 1000 mg-L-1, it flowered 8 d

later.









Delayed flowering in angelonia is consistent with reports of delayed flowering in other

ornamental annuals and perennials sprayed with ethephon (Hayashi et al., 2001; Starman et al.,

2004). This delay in flowering will inhibit the use of ethephon as a PGR during crop production.

However, it will be beneficial in stock production, a production situation where it is

advantageous to maintain plants in a vegetative state, especially those that are day-neutral.

Conclusions

Paclobutrazol drenches, daminozide sprays, and ethephon sprays are effective for

controlling stem elongation in angelonia cultivars. Paclobutrazol drenches are active at very low

concentrations while daminozide and ethephon sprays require much higher concentrations for

efficacy. Response to these PGRs varies with cultivar. For example, 'Angelina Violet and

White' was responsive to paclobutrazol, daminozide and ethephon, while 'AngelMist Dark

Lavender' was responsive to paclobutrazol but less sensitive to ethephon and daminozide sprays.

Sensitivity to the PGRs was cultivar-specific, but was not related to plant vigor. For example,

plants of 'AngelMist Purple Stripe', one of the most vigorous cultivars in the experiments,

drenched with paclobutrazol had a greater percentage of height control related to untreated

plants, while 'AngelMist Purple Improved', a less vigorous cultivar relative to other angelonia

cultivars, had a smaller percentage of height control.

The recommended paclobutrazol drench concentration of 5 mg-L-1 applied two weeks

after transplant (PanAmerican Seed, 2006) was higher than necessary for sufficient control of

plant height in Expt. 5-1. However, it was not sufficient for height control in Expt. 5-3,

primarily due to it being applied too late in production to control overall plant growth. Drenches

should be applied early to mid-way through the crop, before plants start growing rapidly. Early

applications will also help to reduce the amount of lodging present in the very vigorous cultivars,

such as 'AngelMist Purple Stripe' and 'Angelface Blue.' Once plants reach visible bud, it is too









late for drench applications to be effective. These experiments were conducted in a warm

climate during the summer, and lower concentrations should be effective in cooler temperatures.

The recommended rate for daminozide was effective for 'Angelina Violet and White,' the

cultivar for which it was recommended (Selecta First Class, 2008). However, most of the other

cultivars were unresponsive or only slightly responsive to the daminozide sprays. It will need to

be applied at higher concentrations or in a tank mix combination with another chemical for

sufficient efficacy.

Ethephon reduced stem elongation in angelonia cultivars, and the end result was a

marketable plant. However, applications resulted in phytotoxicity symptoms in all cultivars.

The degree of severity varied depended upon cultivar at the low concentrations, but all were

severely damaged at 1000 mg-L-1. 'AngelMist Dark Lavender' and 'Angelface White Improved'

were more sensitive than 'AngelMist Purple Improved' or 'AngelMist Purple Stripe.' Ethephon

helped increase lateral branching, a positive result in cultivars that have strong apical dominance,

but delayed flowering, a negative result in commercial production.


















AM Dk Lav AM Dk Lav Ang Violet & Wht Ang Violet &Wht
-AM Purp Imp AM Purp Imp -AF Blue AF Blue
-AM Lay Stripe A AM Lay Stripe AF White AF White
-AM Purp Stripe AM Purp Stripe
65

55

E 45 A

S35

.z 25

15

5

-5 ,
0 5 10 15 20 A
Paclobutrazol concentration (mg*L"1)

AM Dk Lav AM Dk Lay Ang Violet & Wht Ang Volet & Wht
--AM Purp Imp AM Purp Imp -AF Blue AF Blue
--AM La Stripe A AM Lay Stripe AF White AF White
--AM Purp Stripe AM Purp Stripe
65

55

E 45
6 -35------.-


S25
E
S15

5

-5
0 1250 2500 3750 5000 B
Daminozide concentration (mg*L 1)

AM Dk Lav AM Dk Lay Ang Violet & Wht Ang Volet & Wht
--AM Purp Imp AM Purp Imp -AF Blue A AF Blue
--AM La Stripe A AM Lay Stripe AF White AF White
-AM Purp Stripe AM Purp Stripe
65 -

55.

454



25
E
15

5

-5
0 250 500 750 1000 C
Ethephon concentration (mg*L"1)

Figure 5-1. Effect of plant growth regulators on stem elongation in Angelonia angustifolia 23

days after treatment. Stem elongation was calculated as the difference between final

and initial height. Regression equations (Table 5-1) were calculated using individual

measurements of all plants (n=6). Observed points at each concentration represent

the mean of each cultivar. Plant growth regulators used were A) paclobutrazol B)

daminozide C) ethephon.











117























SA B C D














E F G H
Figure 5-2. Examples of the response of Angelonia angustifolia cultivars to increasing concentrations of paclobutrazol (mg-L-1) (Expt.
5-1). The top row is 'AngelMist Purple Stripe,' a less sensitive cultivar. A) 0 B) 5 C) 10 D) 20 mg-L-1. The bottom row is
'Angelina Violet and White,' a very sensitive cultivar. E) 0 F) 5 G) 10 H) 20 mg-L1.























A B C














E F G H
Figure 5-3. Examples of the response of Angelonia angustifolia cultivars to increasing concentrations of daminozide (mg-L-) (Expt.
5-1). The top row is 'Angelface White,' a less responsive cultivar: A) 0 B) 1250 C) 2500 D) 5000 mg-L1. The bottom row
is 'AngelMist Lavender Stripe,' a responsive cultivar: E) 0 F) 1250 G) 2500 H) 5000 mg-L1.

















\ D


F I


E F G H
Figure 5-4. Examples of the response of Angelonia angustifolia cultivars to increasing concentrations of ethephon (mg-L1) (Expt.
5-1). The top row is 'AngelMist Dark Lavender', a less responsive cultivar: A) 0 B) 1250 C) 2500 D) 5000 mg-L-1. The
bottom row is 'AngelMist Purple Stripe', a responsive cultivar: E) 0 F) 1250 G) 2500 H) 5000 mg-L1.












AM Dk Lav
AM Dk Lav


AM Purp Imp AM Purp Stripe
AM Purp Imp U AM Purp Stripe


25


.. 20
0
I 15
_o
0

E 10
S3


AF White Imp
AF White Imp


Concentration (mg.L"1)

Figure 5-5. Stem elongation in Angelonia angustifolia cultivars treated with ethephon (Expt.
5-4). Data collected at 4WAT. Regression equations (Table 5-6) were calculated
using individual measurements of all plants. Observed points at each concentration
represent the mean of each cultivar (n=9).



AM Dk Lay AM Purp Imp AM Purp Stripe AF White Imp
AM Dk Lav AM Purp Imp U AM Purp Stripe AF White Imp

100

90

80

70 -



0 50-
0
S40

o 30


Concentration (mg.L"1)


Figure 5-6. Days to first open flower in Angelonia angustifolia cultivars treated with ethephon
(Expt. 5-4). Regression equations (Table 5-6) were calculated using individual
measurements of all plants. Observed points at each concentration represent the
mean of each cultivar (n=9).










Table 5-1. Regression equations for stem elongation for Angelonia angustifolia cultivars in response to PGR concentration (Expt.
5-1). Analysis of variance for treatment, cultivar, and treatment x cultivar were significant at P<0.001. Regression
equations were linear (L) or quadratic (Q) and significant at P<0.05, 0.01, or 0.001.


Variable
Paclobutrazol


Daminozide








Ethephon


Cultivar
AVW
AMDL
AMPI
AMLS
AMPS
AFW
AFB

AVW
AMDL
AMPI
AMLS
AMPS
AFW
AFB

AVW
AMDL
AMPI
AMLS
AMPS
AFW
AFB


Significance
L***, Q***
L***, Q***
L***, Q***
L***, Q***
L***, Q***
L***, Q***
L***, Q***


L**, Q*
L***
L***
L*
L***, Q*
L*
L***

L**
L***
L***
L**
L***
L*
L***, Q*


r_
0.94
0.96
0.85
0.92
0.94
0.83
0.89

0.40
0.53
0.53
0.45
0.53
0.24
0.47

0.34
0.46
0.41
0.31
0.73
0.18
0.71


Equation
y = 47.3 -
y =36.8-
y = 39.6 -
y = 44.7 -
y = 58.5 -
y = 54.9 -
y = 35.7 -


z *, *** significant at P<0.05, 0.01, and 0.001, respectively.


47.7
37.9
39.5
45.3
59.3
55.2
38.8

46.1
39.6
37.6
46.3
59.2
54.1
36.1


7.10x + 0.246x2
3.09x + 0.083x2
3.03x + 0.078x2
4.74x + 0.140x2
5.74x + 0.164x2
5.31x + 0.158x2
4.37x + 0.144x2


0.008x + 0.000001x2
0.OO1x
0.002x
0.002x
0.005x + 0.0000005x2
0.003x
0.003x

0.01x
0.01x
0.01x
0.01x
0.03x
0.01x
0.04x + 0.00002x2










Table 5-2. Stem elongation (cm) in Angelonia angustifolia four weeks after application of
daminozide (Expt. 5-2). The cultivar x concentration interaction was non-significant.
Cultivar and concentration were significant at P<0.001 respectively. This table
presents the main effects of cultivar and concentration. Same letters within each row
or column are not significantly different using Tukey's at a=0.05.
Daminozide concentration (mg-L1)
Cultivar 0 1250 2500 5000 Pooled mean (cv)
AMDL 46.7 41.6 38.3 35.2 40.5 c
AMPS 88.5 76.8 77.0 72.2 78.6 a
AFW 52.6 46.3 44.8 38.4 45.5 b


Pooled mean
(Daminozide)


62.6 a


54.9 b


53.4 b


49.0 c


Table 5-3. Stem elongation (cm) in Angelonia angustifolia in response to paclobutrazol
applications (Expt. 5-3). The early spray was applied 2 weeks after pinch. The late
drench was applied at visible bud stage, at 4 weeks after pinch for 'AngelMist Purple
Stripe' and at 6.5 weeks for 'Angelface White'. Mean separation was performed
using Tukey's at a=0.05.
Treatmenty Concentration AngelMist Purple Angelface White


Late drench
Late drench
Late drench
Early spray
Early spray
Control
Pooled means


2
4
8
50
100


Stripe
89.3
85.8
81.2
85.8
81.9
83.0
84.5 a


72.4
64.6
61.6
70.9
64.0
65.7
66.5 b


Table 5-4. Analysis of variance for Angelonia angustifolia treated with ethephon (Expt. 5-4).
For each treatment, n=12.
Phytotoxicity Stem Lateral Time to
elongation number flower
Cultivar *** *** *** ***
Concentration *** *** ***
Cv. xconc. *** *** NS ***
NS,*, **, *** Nonsignificant or significant at P<0.05, 0.01, and 0.001, respectively.


t_>









Table 5-5. Mean separation for severity of phytotoxicity symptoms in Angelonia angustifolia
one week after treatment with ethephon (Expt. 5-4). Phytotoxicity was rated on a 0 -
3 scale, with 0 = no visible symptoms, 1 = slight chlorosis of leaf tips, 2 = moderate
chlorosis of leaves and slight necrosis at leaf edges, and 3 = severe chlorosis of
leaves, moderate to severe leaf necrosis, and leaf distortion. Mean separation was
performed using Tukey's at a=0.05, and HSDo 005 = 0.6.
Cultivar
Ethephon AngelMist AngelMist AngelMist Angelface
(mg-L1) Dark Lavender Purple Improved Purple Stripe White
0 0.0 c 0.0 c 0.0 c 0.0 c
250 0.1 c 0.1 c 0.3 c 0.2 c
500 2.7 a 1.8 b 1.4 b 2.9 a
1000 2.9 a 2.7 a 2.0 b 2.9 a

Table 5-6. Regression equations for Angelonia angustifolia cultivars treated with ethephon
(Expt. 5-4).
Variable Cultivar Significancez r2 Equation
2
Stem elongation (cm) AMDL L***, Q*** 0.78 y = 25.8 0.03x + 0.00002x
AMPI L** 0.29 y = 12.6- 0.005x
AMPS L***, Q** 0.78 y = 24.8 0.03x + 0.00002x2
AFWI L***, Q** 0.59 y = 11.0 0.01x + 0.00001x2

Days to flower AMDL L***, Q*** 0.80 y = 33.4 + 0.06x 0.00004x2
AMPI L*** 0.36 y = 66.8 + 0.0124x
AMPS L***, Q** 0.68 y = 54.7 + 0.03x 0.00002x2
AFWI L*** 0.67 y = 65.7 + 0.02x
zNS,, *, *** Non-significant or significant at P<0.05, 0.01, and 0.001, respectively.

Table 5-7. Mean separation for number of laterals in Angelonia angustifolia 4 weeks after
ethephon treatment (Expt. 5-4). Mean separation was performed using Tukey's at a=0.05.
HSD values for the main effects of both cultivar and concentration were 1.7. Same
letters within each row or column are not significantly different.


Ethephon concentration (mg-L )
0 250 500 1000 Pooled means (cv)
19.8 20.7 20.7 22.7 20.9 a
3.8 4.0 5.1 7.0 4.8b
6.3 4.2 4.6 6.3 5.4b
3.4 3.9 5.8 6.3 4.9 b


ans


9.1 ab


Cultivar
AMDL
AMPI
AMPS
AFWI
Pooled me
(ethephon)


8.3 b


8.2 b


10.8 a









CHAPTER 6
CONCLUSIONS

The increase in popularity of Angelonia angustifolia has led to the development and

release of cultivars with novel growth habits and flower color, and companies are including these

new releases within their available product line, often referred to as a series. The availability of

new cultivars into an existing product line does not guarantee that all of them will have similar

growth habits or production requirements, which can be a problem for growers. In production, it

is optimal to select cultivars with similar production requirements, such as optimal fertilizer

concentration and like sensitivities to PGRs. Also, time to flower will determine the length of

the production schedule, and growers would prefer cultivars with shorter production times so that

more turns can be accomplished in a greenhouse during a season.

Thirty-one Angelonia angustifolia cultivars were observed for differences in growth habit

and flowering traits. Differences of more than 3 weeks were observed between cultivars for

number of days to first flower and number of days to marketability. Plant height at marketability

ranged from 14 to 64 cm, and plant width ranged from 28 to 54 cm. Cultivars with naturally tall

or wide growth habits will be difficult to grow in production without the use of PGRs to inhibit

stem elongation. Another issue in production is the susceptibility of certain cultivars to powdery

mildew. Plants were grown in a naturally ventilated greenhouse and not treated with any

fungicides for the duration of the experiment. In some cultivars, no powdery mildew was

detected, while in others, it severely affected plant quality. The range of susceptibility observed

may indicate that it could be a trait that breeders can select for in their breeding programs.

Angelonia is a warm season crop produced during the early spring in southern states and

in late spring for the northern states. While angelonia are not photoperiodic, they respond to

changes in temperature and light quantity associated with seasonality. From late winter to early









summer, cultivars flowered 17 to 29 d earlier, indicating that temperature and light most likely

play a major role in hastening flower development. In cooler climates, the cost of supplemental

heating and lighting may be cost-effective if the crop time can be shortened significantly.

Angelonia cultivars responded similarly to increasing applications of fertilizer and can be

treated uniformly in a greenhouse. This is beneficial to large-scale growers who may have

multiple annual species or cultivars in a greenhouse section. They respond well to a complete

fertilizer with N at 100 mg-L-1, which is similar to many other warm season annuals (Kang and

van lersal, 2002; Kent and Reed, 1996). Although this is less than recommended for other crops,

such as petunia (James and van lersal, 2002), cultivars did not show any sensitivity to increased

fertilizer concentrations other than the appearance of darker green leaves and a slight reduction

in growth.

Angelonia cultivars, however, are cultivar-specific in response to PGRs. Daminozide is

effective at 5000 mg-L-1 in 'Angelina Violet and White' and 'Angelface Blue' but other cultivars

are relatively insensitive. Paclobutrazol is ineffective as a spray, but is highly effective as a

drench if applied early in the crop. Ethephon is effective at inhibiting stem elongation, but the

potential for phytotoxicity is greater in certain cultivars. None of the cultivars show permanent

damage from ethephon, but its ability to delay flowering can be an issue.

Angelonia cultivars show a wide range of heat tolerance with respect to flowering during

the summer months. The earliest plant date, 4 Apr. 2007, provided the best display of color

when all of the cultivars reached their peak flowering ability. In the latest plant date, none of the

cultivars were able to get established adequately and provide a similar display of color. All of

the cultivars were negatively affected by the high temperatures, but 'AngelMist Purple Stripe'

and 'Angelface White' were less affected.









In greenhouse production, 'AngelMist Dark Lavender' is an ideal cultivar to grow due to

its early flowering and compact plant habit, which requires minimal growth regulation to keep its

size under control. However, it is very sensitive to powdery mildew and will need to be

monitored during the spring growing season. In the landscape, it will flower in late spring and

early summer, but not stay in flower throughout the summer. This is an example of a

greenhouse-friendly cultivar but not a landscape-friendly cultivar. On the other side of the

spectrum is 'AngelMist Purple Stripe.' It is a medium to late flowering cultivar, and it is one of

the most vigorous angelonia cultivars available. It also has very sticky, oily, pubescent leaves

and stems, which makes the application of PGRS, which are necessary to keep it from becoming

overgrown, difficult. However, in the landscape, it has excellent heat tolerance and will provide

a consistent display of color throughout the summer season.

Our study complements the studies published by Miller and Armitage (2002) and Miller

et al. (2001) on angelonia production. However, many gaps still exist, including the specific

effects of temperature and light on flowering. What is the relative impact of temperature and

flowering on flower initiation and development, and can this be quantified such that a grower can

know exactly how to hasten or delay marketability of the crop? In certain cultivars, such as

'AngelMist Purple Stripe' and 'Angelface White', it appears that certain traits may be linked; for

example, these were the only cultivars with sticky pubescent leaves and also they were the only

two to continuously flower throughout the summer, which may indicate heat tolerance relative to

other cultivars with respect to floral development. Also, 'Angelface White' was the latest

cultivar to flower and 'AngelMist Purple Stripe' was one of the later-flowering cultivars,

possibly indicating that these two may have a higher temperature optimum for flower

development and continued flowering.









Angelonia is a crop with breeding potential. The range of variability present within this

species indicates that many of these traits can be selected and improved upon such that a cultivar

that is easily grown and produced and will provide good landscape color during the season. In

addition, A. angustifolia is the only species known to currently be in commercial production and

related species may hold cultivation potential or have desirable traits which could be

incorporated into a breeding program.









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BIOGRAPHICAL SKETCH

Jennifer Kay Boldt was born in Minnesota and raised in Florida. She and her twin sister

Jessica embraced horticulture at a young age and grew up helping their parents in the family-

owned nursery and garden center. She earned a Bachelor of Science in environmental

horticulture and a Bachelor of Arts in business administration from the University of Florida in

December 2005. As an undergraduate, Jennifer completed two horticulture-related internships: a

six-month production internship with Van Wingerden International in Fletcher, North Carolina,

and a three-month trial garden internship with Ball Horticultural Company in West Chicago,

Illinois. As a graduate student, she was a research assistant and coordinated the University of

Florida's on-campus floriculture trial gardens. Jennifer's future career plans lead her back to

Minnesota, where she will begin a PhD program in Fall 2008.





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1 CULTURAL AND ENVIRONMENTAL FACTOR S INFLUENCE THE PERFORMANCE OF Angelonia angustifolia CULTIVARS By JENNIFER KAY BOLDT A THESIS PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLOR IDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE UNIVERSITY OF FLORIDA 2008

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2 2008 Jennifer Kay Boldt

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3 To Mom and Dad

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4 ACKNOWLEDGMENTS I would like to thank m y advisor and committee chair, Dr. James Barrett, for all of his guidance and support during my years as an undergra duate and graduate stude nt at the University of Florida. Without his nudging, I may not have decided to pursu e a masters program. During the past two and a half years, he listened to all of my ideas (b oth the good and the bad), provided unwavering support for my research and kept me well-fed during long days of data collection and trial garden planting. I w ould also like to thank my comm ittee members, Drs. James Gibson, Rosanna Freyre, and Kenneth Quesenbe rry, for their support and guidance. I thank Ms. Carolyn Bartuska for patientl y answering all of my many statistical questions. More importantly, she managed to su rvive working in the same office as my twin sister and me, which was no easy task when we both got stressed! Weekly lunch outings somehow managed to keep us all sane. Thanks also go to Bob Weidma n for taking care of my research plants when I was out of town; without alive plants, I would not have had any results. Much appreciation is given to my twin sister Jessica for continually offering to help me collect data in a hot, humid greenhouse when no one else was available to help. She was always the behind-the-scenes person maki ng sure everything was taken care of so that I could finish experiments and write this thesis. Finally, I would like to thank my parents for all of their support and guidance. They provided me with my first glimpse into the worl d of floriculture, taugh t me the importance of hard work and finishing a task, and patiently listened to all of my joys and frustrations.

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5 TABLE OF CONTENTS page ACKNOWLEDGMENTS...............................................................................................................4 LIST OF TABLES................................................................................................................. ..........8 LIST OF FIGURES.......................................................................................................................10 LIST OF FIGURES.......................................................................................................................10 ABSTRACT...................................................................................................................................12 CHAP TER 1 LITERATURE REVIEW.......................................................................................................14 Introduction................................................................................................................... ..........14 Angelonia................................................................................................................................15 Taxonomy........................................................................................................................15 Botany..............................................................................................................................15 Pollination and Reproduction.......................................................................................... 16 Described Species............................................................................................................ 17 Cultivation.......................................................................................................................18 Commercial Cultivars...................................................................................................... 19 Flowering................................................................................................................................19 Irradiance Influences Time to Flower.............................................................................21 Temperature Influences Time to Flower......................................................................... 22 Temperature x Irradiance Interactions Influence Time to Flower................................... 23 Flowering in Angelonia...................................................................................................24 Factors Influencing Continued Flowering....................................................................... 25 Plant Growth...........................................................................................................................27 Temperature.................................................................................................................... .27 Irradiance.........................................................................................................................28 Temperature x Irradiance Interactions.............................................................................29 Fertilizers................................................................................................................................30 Plant Growth....................................................................................................................30 Use of a Hand-held Meter to Measure Chlorophyll Content ..........................................31 Plant Growth Regulators.........................................................................................................33 Introduction................................................................................................................... ..33 Paclobutrazol...................................................................................................................34 Daminozide......................................................................................................................37 Ethephon..........................................................................................................................38 Objectives...............................................................................................................................41

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6 2 Angelonia angustifolia CULTIVAR SCREEN ...................................................................... 44 Introduction................................................................................................................... ..........44 Materials and Methods...........................................................................................................44 Experiment 2-1 Cultivar Screen of 31 Comm ercial Cultivars........................................ 44 Experiment 2-2 Cultivar Comparison of Seven Cultiv ars across Multiple Seasons....... 46 Results and Discussion......................................................................................................... ..47 Experiment 2-1................................................................................................................ 47 Days to first flower...................................................................................................47 Days to marketability...............................................................................................48 Plant height and widt h at m arketability.................................................................... 49 Inflorescence number at marketability..................................................................... 50 Inflorescence number at 9.5 weeks.......................................................................... 51 Flower size............................................................................................................... 51 Powdery mildew index............................................................................................. 52 Experiment 2-2................................................................................................................ 53 Days to first flower...................................................................................................53 Plant height...............................................................................................................55 Flower size............................................................................................................... 56 Powdery mildew susceptibility................................................................................ 57 Conclusions.............................................................................................................................57 3 PLANT DATE x CULTIVAR INTERACTIONS INFLUENCE SUMMER LANDSCAPE PERF ORMANCE OF Angelonia angustifolia ...............................................69 Introduction................................................................................................................... ..........69 Materials and Methods...........................................................................................................69 Results and Discussion......................................................................................................... ..71 10-Week Data..................................................................................................................71 Plant height...............................................................................................................71 Inflorescence number...............................................................................................72 Flower height............................................................................................................73 Flower Ratings by Plant Age...........................................................................................74 Flower Ratings by Evaluation Date.................................................................................75 Conclusions.............................................................................................................................77 4 CULTIVAR BY FERTILIZER INTERA C TIONS AFFECT GROWTH OF Angelonia angustifolia .............................................................................................................................83 Introduction................................................................................................................... ..........83 Materials and Methods...........................................................................................................84 Results and Discussion......................................................................................................... ..85 Substrate pH and Electrical Conductivity....................................................................... 85 Leaf Greenness................................................................................................................87 Plant Growth....................................................................................................................89 Flowering.........................................................................................................................90 Conclusions.............................................................................................................................91

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7 5 Angelonia angustifolia CULTIVARS DIFFER I N RESPONSE TO PACLOBUTRAZOL, DAMINOZIDE, AND ETHEPHON................................................ 102 Introduction................................................................................................................... ........102 Materials and Methods.........................................................................................................103 Experiment 5-1 Paclobutrazol, Dami nozide, and Ethephon Response Curves ............. 103 Experiment 5-2 Daminozide.......................................................................................... 104 Experiment 5-3 Paclobutrazol Sp ray and Drench Applications .................................... 105 Experiment 5-4 Ethephon.............................................................................................. 105 Results and Discussion......................................................................................................... 107 Experiment 5-1.............................................................................................................. 107 Paclobutrazol..........................................................................................................107 Daminozide............................................................................................................108 Ethephon.................................................................................................................109 Experiment 5-2.............................................................................................................. 109 Experiment 5-3.............................................................................................................. 111 Experiment 5-4.............................................................................................................. 112 Phytotoxicity..........................................................................................................112 Stem elongation...................................................................................................... 113 Lateral number.......................................................................................................114 Flowering...............................................................................................................114 Conclusions...........................................................................................................................115 6 CONCLUSIONS.................................................................................................................. 125 LIST OF REFERENCES.............................................................................................................129 BIOGRAPHICAL SKETCH.......................................................................................................139

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8 LIST OF TABLES Table page 1-1 Current taxonomi c class ification of Angelonia angustifolia .............................................42 1-2 Commercially availa ble angelonia series........................................................................... 42 1-3 Cultivar names of angel onia used in experim ents............................................................. 43 2-1 Plant growth and flowering characteristics for seven Angelonia angustifolia cultiva rs used in Expt. 2-2................................................................................................................61 2-2 Temperature and light data for Expt. 2-2. .......................................................................... 62 2-3 Angelonia a ngustifolia flowering data for 31 cultivars (Expt. 2-1)................................... 63 2-4 Plant data for 31 cultivars of Angelonia angustifolia collected at m arketability (Expt. 2-1).....................................................................................................................................64 2-5 Angelonia a ngustifolia plant data for 31 cultivars co llected 9.5 week s after pinch (Expt. 2-1).................................................................................................................... ......65 2-6 Split plot analysis of variance for seven Angelonia angustifolia cultivars grown at four different seasons from Ja nuary to July 2007 (Expt. 2-2). .......................................... 66 2-7 Number of days to first flower in seven Angelonia angustifo lia cultivars across four seasons (Expt. 2-2)............................................................................................................ .66 2-8 Plant height (cm) at m arketability in seven Angelonia angustifolia cultivars across four seasons (Expt. 2-2)..................................................................................................... 67 2-9 Flower height (mm) in six Angelonia angustifolia cultivars across four seasons (Expt. 2-2). ................................................................................................................... ......67 2-10 Powdery mildew index in seven Angelonia angu stifolia cultivars across four seasons (Expt. 2-2).................................................................................................................... ......68 3-1 Monthly temperature and DLI data for Gainesville, FL for summ er 2007....................... 80 3-2 Split-plot analysis of variance for An gelonia angustifolia growth and flowering data collected at 10 weeks after pl anting for three plant dates.................................................. 80 3-3 Mean separation for plant height (cm) of eight An gelonia angustifolia cultivars collected at 10 weeks after tran splant for three plant dates............................................... 81 3-4 Mean separation for number of inflorescences per plant of eig ht Angelonia angustifolia cultivars collected at 10 weeks after transplant for three plant dates............ 81

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9 3-5 Mean separation for flower height (mm) of eight Angelonia a ngustifolia cultivars collected at 10 weeks after tran splant for three plant dates............................................... 81 3-6 Split-plot analysis of variance for An gelonia angustifolia flower ratings by plant age (weeks after transplant)......................................................................................................82 3-7 Split-plot analysis of variance for An gelonia angustifolia flower ratings by evaluation date...................................................................................................................82 4-1 Analysis of variance for Angelonia a ngustifolia cultivars in response to fertilization...... 98 4-2 Regression equations for pH and EC m easurem ents collected at 6 weeks after treatment...................................................................................................................... ......98 4-3 Regression equations for SPAD readings collected at 6 weeks after treatm ent................ 99 4-4 Regression equations for plant size, dry weight, and num ber of inflorescences at 6 weeks after treatment....................................................................................................... 100 4-5 Number of days to first open flower in Angelonia angustifo lia ......................................101 5-1 Regression equations for stem elongation for An gelonia angustifolia cultivars in response to PGR concentration (Expt. 5-1)..................................................................... 122 5-2 Stem elongation (cm) in Angelonia a ngustifolia four weeks after application of daminozide (Expt. 5-2).................................................................................................... 123 5-3 Stem elongation (cm) in Angelonia a ngustifolia in response to paclobutrazol applications (Expt. 5-3).................................................................................................... 123 5-4 Analysis of variance for Angelonia a ngustifolia treated with ethephon (Expt. 5-4)....... 123 5-5 Mean separation for severity of phytotoxicity sym ptoms in Angelonia angustifolia one week after treatment with ethephon (Expt. 5-4)........................................................ 124 5-6 Regression equations for Angelonia angustifo lia cultivars treated with ethephon (Expt. 5-4).................................................................................................................... ....124 5-7 Mean separation for number of laterals in Angelonia angustifolia 4 weeks after ethephon treatm ent (Expt. 5-4)........................................................................................ 124

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10 LIST OF FIGURES Figure page 2-1 Range of plant height present in commercial Angelonia angu stifolia cultivars................ 59 2-2 Examples of the range on inflorescence number present in A ngelonia angustifolia cultivars 9.5 weeks after pinching..................................................................................... 59 2-3 Range of flower size present in commercial Angelonia angu stifolia cultivars................. 60 2-4 Comparison between inflorescences in Angelonia angustifolia Angelf ace White and Serena White............................................................................................................ 60 3-1 Flower ratings of Angelonia angustifolia graphed by weeks after transplant (W AT)....... 78 3-2 Flower ratings of Angelonia angustifolia graphed by date of data collection. ..................79 4-1 Effect of increasing fertilization with 20.0N-4.4P-16.6K on substrate pH in Angelonia a ngustifolia 6 weeks after start of treatments...................................................93 4-2 Effect of increasing fertilization w ith 20.0N-4.4P-16.6K on s ubstrate electrical conductivity in Angelonia angustifolia 6 weeks after start of treatm ents.......................... 93 4-3 Effect of increasing fertilization with 20.0N-4.4P-16.6K on lower leaf SPAD values in Angelonia angustifolia 6 weeks after start of treatm ents............................................... 94 4-4 Effect of increasing fertilization w ith 20.0N-4.4P-16.6K on upper leaf SPAD values in Angelonia angustifolia 6 weeks after start of treatm ents............................................... 94 4-5 Response of Angelonia a ngustifolia AngelMist Dark Lavender to increasing rates of fertilization with 20.0N-4.4P-16.6K..............................................................................95 4-6 Effect of increasing fertilizati on with 20.0N-4.4P-16.6K on plant size in A ngelonia angustifolia 6 weeks after start of treatments.................................................................... 96 4-7 Effect of increasing fertilization w ith 20.0N-4.4P-16.6K on plant dry weight in Angelonia a ngustifolia 6 weeks after start of treatments...................................................96 4-8 Effect of increasing fertilization with 20.0N-4.4P-16.6K on number of inflorescences in Angelonia angustifolia 6 weeks after start of treatm ents............................................... 97 5-1 Effect of plant growth regulators on stem elongation in Angelonia angustifolia 23 days after treatm ent.......................................................................................................... 117 5-2 Examples of the response of Angelonia angustifolia cultiva rs to increasing concentrations of paclobutrazol (mgL-1) (Expt. 5-1)...................................................... 118

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11 5-3 Examples of the response of Angelonia angustifolia cultiva rs to increasing concentrations of daminozide (mgL-1) (Expt. 5-1)......................................................... 119 5-4 Examples of the response of Angelonia angustifolia cultiva rs to increasing concentrations of ethephon (mgL-1) (Expt. 5-1)............................................................. 120 5-5 Stem elongation in Angelonia angustifolia cultivars treated with ethephon (Expt. 5-4). ..................................................................................................................................121 5-6 Days to first open flower in Angelonia angustifolia cultivars treated with ethephon (Expt. 5-4) ........................................................................................................................121

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12 Abstract of Thesis Presen ted to the Graduate School of the University of Florida in Partial Fulfillment of the Requirements for the Degree of Master of Science CULTURAL AND ENVIRONMENTAL FACTOR S INFLUENCE THE PERFORMANCE OF ANGELONIA ANGUSTIFOLIA CULTIVARS By Jennifer Kay Boldt August 2008 Chair: James Barrett Major: Horticultural Sciences Thirty-one commercial Angelonia angustifolia Benth. cultivars were evaluated for phenotypic differences in growth and floweri ng. Number of days to first flower and marketability during spring production ranged from 41 to 64 d and 45 to 70 d, respectively. At marketability, plant height ranged from 14 to 64 cm, plant width from 28 to 54 cm, and inflorescence number from 2.3 to 4.2. At 9.5 w eeks after pinch, inflorescence number ranged from 2.1 to 29.2. Flower size ranged from 20.1 to 30.7 mm tall and 12.7 to 28.4 mm wide. Cultivars differed in powdery mildew sensitivity. In a sub-sample of eight cultivars, a season x cultivar interaction was significan t for all parameters except flower size. Time to first flower decreased and plant height increased from s eason 1 to 4, but the magnitude differed with cultivar. Eight cultivars planted in a landscape bed at three intervals differed significantly in summer performance. Ten weeks after transpla nt, the interaction was significant for plant height, inflorescence number, and flower size. Decreased summer flowering was influenced by environmental conditions rather than plant age. Summer flowering wa s poor in all cultivars except AngelMist Purple Stripe.

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13 Cultivars subirrigated with increasing fertilizer concentrations exhibited an interaction for all parameters except final pH and number of days to first flower, which were due to the main effect of fertilizer concentration and cultiv ar, respectively. SPAD values increased as N concentration increased. EC increased and pH d ecreased as N concentration increased. Optimal plant size, dry weight, and inflorescence number o ccurred at different concentrations. Responses to fertilization were not cultiv ar specific and recommended concentrations are at N levels of 100 mg/L or slightly higher. Cultivars showed varying sensitivities to paclobutrazol, daminozide, and ethephon. All effectively inhibited growth in at least one cu ltivar, and paclobutrazol drenches inhibited stem elongation in all cultivars. P aclobutrazol drenches of 5 mg/L applied 2 weeks after pinch inhibited stem elongation, but spra ys of 100 mg/L applied 2 weeks after pinch and drenches of 8 mg/L applied at visible bud were ineffective. Ethephon inhibited stem elongation, increased lateral branching, delayed flow ering, and caused phytotoxicity.

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14 CHAPTER 1 LITERATURE REVIEW Introduction The floriculture industry in the late 1980s sa w a rise in popularity of new, vegetativelypropagated annual crops. Consum er interest in new crops and a shift towards container gardening has helped vegetative annuals become an important product for the industry. In a 15-state survey of growers havi ng at least $10,000 in sa les, their total crop value in 2007 was $4.1 billion and bedding plants accounted for $1.76 billion of the total (NASS, 2008). Approximately 50% of the bedding plants sold were classified as other and not as one of the staple landscape crops: begonias, ge raniums, impatiens, marigolds, pa nsies, petunias, or violas. This other group is a combination of seed a nd vegetatively-propagated crops, but it reflects consumer preference towards selecting a combina tion of older, stand-by crops and newer, lesser known crops for gardens. To catch a consumers attention in the garden center and entice him or her to purchase a particular plant, a new crop must have some tra it that makes it stand out from the others. Thus, breeders have focused on developing new cultivar s with novel growth habits, flower forms, and/or flower colors. This is most eas ily done through the devel opment of vegetativelypropagated crops due to the quicker selection cycle needed relati ve to developing uniform seedproduced cultivars. Angelonia angustifolia a vegetatively-propagated or s eed-propagated crop, is available from multiple young plant suppliers. Within the ra nge of cultivars available from each company, commonly referred to as a series, there are differen ces in growth habit and flowering time. This can lead to problems in production, where sim ilar schedules and produc tion requirements are ideal for large-scale production. Having a range of variability within a species can be both

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15 positive and negative; it provides the consumer with options at retail, but it can make it difficult for a grower to efficiently produce all of the cultivars. Thus a dilemma arises during new cultivar selection as to whether or not to releas e a cultivar and place it w ithin an existing series even if it is not uniform with th e rest of the other cu ltivars already present in that collection. Currently, the range of variab ility present in angelonia cu ltivars is unknown, both between different series and within a series. Within the variability present, are ther e certain traits that are beneficial to this crop or are there traits that ar e hindering its success in production and in the landscape? It will be benefici al to quantify the variability present and determine if current production recommendations are crop-s pecific or cultivar-specific. Angelonia Taxonomy Angelonia H umb. & Bonpl., a genus containing appr oximately 30 species, is native to Mexico, Central and South America, and the We st Indies (Bailey and Bailey, 1976; Huxley, 1992). It originally was placed in Scrophularia ceae (complete taxonomic classification found in Table 1-1), but taxonomists have reorganized this family in recent years and currently place Angelonia in Plantaginaceae (Albach et al., 2005). It is diploid with a base chromosome number of 10 (2n=2x=20) (Steiner, 1996). Botany Angelonia species are subshrubs or perennial herbs and range in height from 0.3 to 0.8 m tall (Bailey and Bailey, 1976). Leav es are opposite in orientation, and may be either glabrous or pubescent (Bailey and Bailey, 1976; Huxley, 1992). The flowers, borne by themselves in leaf axils or on a terminal raceme inflorescence (H uxley, 1992), are zygomorphic with a two-lipped corolla. The upper lip is two-lobed and the lo wer lip is three-lobed (Griffiths, 1994). The corolla does not contain a spur, like in other closely-related species, and the corolla tube is highly

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16 reduced or absent entirely (Huxley, 1994). The st igma and four anthers are located on the top of the gullet created by the reduced corolla tube (Vogel and Machado, 1991). The corolla tends to be blue, lavender, or white -colored (Griffiths, 1994; Vogel and Machado, 1991). Seeds are produced within a fruit capsule (Bailey and Baile y, 1976), and a distinguishi ng characteristic of this genus is its lack of an endos perm in the seed (Steiner, 1996). Angelonia flowers reward pollinators with oil rather than nectar, first described by Vogel (1974) to exist in 1260 species, 50 genera, and five families: Iridaceae, Krameriaceae, Malphigiaceae, Orchidaceae, and Scrophulariaceae. He used the term elaiophore to describe the glands where the oil is produced. Angeloni a flowers have trichome elaiophores, collections of hundreds of thousands of tiny glands that secrete oil. The oil is absorbed by the tarsal pads of visiting female bees (Kampny, 1995; Vogel, 1974). Bees, specifically Centris species, are the common pollinators of Angelonia and use the oil as a food s ource for larvae (Buchman, 1987; Vogel and Machado, 1991). Pollination and Reproduction The flowers of Angelonia are zygom orphic and designed for insect pollination. Upon alighting, a bee places its front le gs into the flower to collect o il from the pouches. In doing so, its head comes in contact with the re productive organs (Vogel and Machado, 1991). Not surprisingly, due to its close relationship wi th the oil-collecting bees, the majority of Angelonia are outcrossing species. Through a combin ation of field obser vations and greenhouse experiments, Machado et al. (2006) determined that among five species of Angelonia observed from the Caatinga in northeastern Brazil, the f our perennial species were self-incompatible and the annual A. pubescens was self-compatible. Angelonia flowers are protandrous and senesce five to seven days after opening. The anthers dehisce during the second to fourth days after flower opening, and the stigma does not become receptive until the fourth or fifth day (Vogel

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17 and Machado, 1991). In its native habitat, flower ing occurs from February or March until June or July and coincides with the ra iny season (Vogel and Machado, 1991). Described Species Approxim ately 30 species of Angelonia are known,, but very few are mentioned in the literature and even fewer have been cultivated. It is believed that A. angustifolia and A. integerrima are the only cultivated sp ecies (Schoellhorn, 2002). Co mmon names for this genus include angelonia, summer snapdragon, and summ er orchid (Hamrick, 2003). The following are brief descriptions of species mentioned in the literature: A. angustifolia Benth., a perennial species native to Mexico and the West Indies, grows approximately 0.3 to 0.5 m tall and has glabrous stems, lanceolate leaves, and racemes containing deep mauve to violet fl owers. It flowers in the summe r, has been naturalized in many areas of the neotropics, and is the species most widely cultivated in commercial horticulture. Two of the old, non-patented cultivars sold commer cially are Alba, a white-flowered selection, and Blue Pacific, a blue and white flowered selection (Baile y and Bailey, 1976; Griffiths, 1994; Huxley, 1992; USDA, 2006). A. cornigera Hook., an annual species native to Brazil, will grow about 0.3 m tall. Flowers are located in the leaf axils as solitary flowers rather than as an inflorescence (Bailey, 1922). A. gardneri Hook., a perennial species native to Braz il that will grow up to 1 m tall, has glandular-pubescent stems and lanceolate-serrate leaves. It flowers in the summer and has purple flowers with white centers (Griffiths, 1994; Huxley, 1992). A. grandiflora Hort. was first introduced to gardener s in 1897 by Benary (Bailey, 1906). It is believed to be a horticultural selection of A. salicariifolia A. integerrima Spreng., a perennial species native to southern Brazil and Paraguay, has entire, lanceolate leaves and s ky blue flowers containing purple s pots. The upper corolla lip is

PAGE 18

18 reduced and the lower corolla lip has ovate, coarsely-serrated lobes. It flowers in the summer (Griffiths, 1994; Huxley, 1992). A. procumbens a native to eastern Brazil, has not been described in much detail except that it shares similar characteristics with othe r species in this genus (Barringer, 1985). A. salicariifolia Humb. & Bonpl., a perennial species native to Central America and the West Indies, grows approximately 0.5 to 0.8 m (Bailey and Bailey, 1976). It has extremely glandular, sticky-pubescent stems; broad, lanceola te, serrulate leaves; and mauve or bluishpurple flowers. The only described cultivar is Grandiflora (also known as A. grandiflora ), which has large white flowers that can reach 25 mm in size (Bailey and Bailey, 1976; Griffiths, 1994; Huxley, 1992; Pennell, 1920). Cultivation Angelonia is commonly grown outdoors as a pere nnial in tropical a nd subtropical regions and as a summer annual in cooler clim ates (Huxl ey, 1992). It is commonly found growing along damp edges of savannahs or in open, sunny locations in its native habitat. Multiple species are located in the Caatinga, a tropical dry forest in northeastern Brazil, where rainfall is seasonal and temperatures consistently range betw een 23 and 27 C (Machado et al., 1991). The first known reference to its commercial availability is from Benary in 1897 (Bailey, 1906), but it was not widely popularized until the late 1990s. The high incidence of infection with Cucumber Mosaic Virus limited its availability due to a lack of growers willing to grow plants that could be virus inf ected (Hamrick, 2003). In the late 1990s, Ball FloraPlant released multiple cultivars in the AngelMist series. Thes e cuttings were touted as coming from clean stock plants that had undergone virus-indexing and subsequent virus eradication (Schoellhorn, 2002). Since then, multiple companies have released commercial series. A series was a term originally used to denote seed lines offered by one company that were very similar to each other

PAGE 19

19 and varied only by one trait, such as flower color (Armitage, 1994). This term has since been extended to describe a line of vegetatively-pr opagated cultivars available from a company. Commercial Cultivars Comm ercial cultivars are currently available fr om at least six different companies (Table 1-2). All of them are vegetatively-propagated except for the seed-propagated Serena series. Through breeding advances, the rang e of flower colors availabl e include white, pink, lavender, dark blue, purple, and bi-colored. It is a ve rsatile plant and can be used in mass landscape plantings, hanging baskets, and mixe d containers or as a cut flower. Variability is present in angelonia for growth habit and flowering, includi ng plant height, plant width, amount of leaf pubescence, degree of leaf stickiness number of days to first flow er, flower color, flower size, degree of reflexing in the two side lobes of the lower lip, inflorescence length, inflorescence number, disease resistance, and le ngth of flowering in landscape (Boldt, personal observation). Angelonia is a warm season crop grown primarily for the spring and summer seasons in southern states and for the summer season in nor thern states (Armitage, 1997). From a rooted cutting, crop time will range from 6 to 10 weeks depending upon cultivar and greenhouse conditions (Schoellhorn and Alvarez, 2002). Optimal growth occu rs when plants are provided with high irradiance levels, warm temperatures, and moderate ferti lizer concentrations. Cultural problems tend to be few as long as adequate greenhouse conditions are maintained. A more detailed review of factors influencing angel onia production will be discussed in the following sections. Flowering For m ost greenhouse crops, the onset of flower ing is one of the most important steps in determining that it is ready to be shipped to retail stores and gard en centers. In nature, the timing of flowering is important for plants so that they maximize their potential for successful

PAGE 20

20 pollination and future survival (Bernier et al., 199 3). In most species, environmental cues such as photoperiod, temperature, and water availability are major factors that cue the transition from a vegetative phase to a reproductive phase (Berni er et al., 1993). The major genetic pathways controlling flowering in plants are the photoperiodic, vernalizati on, gibberellic acid (GA), and autonomous pathways. Plants are classified into three major groups with respect to thei r photoperiod response: short-day plants flower when the night length is longer than a critical period, long-day plants flower when the night length is shorter than a critical period, and day-neutral plants flower regardless of the length of the night period (Roberts and Summerf ield, 1987). Day neutral plants are also termed photoperiod-insensitive or autonomous-flowering (B ernier et al., 1993; Halevy, 1984). The number of floricultural crops classified as day-neutral is small compared to the number of photoperiodic-sensitive crops. Some of the most im portant crops are photoperiodic, including Euphorbia pulcherrima (poinsettia), Dendranthema x grandiflorum (chrysanthemum), and Kalanchoe blossfeldiana (kalanchoe). In two recent studies, 68 percent of bedding plants and 86 percent of Hibiscus species trialed were photoperi odic (Mattson and Erwin,2005; Warner and Erwin, 2001a). A response to photoperiod, in general, is relate d to the latitude and elevation from which the species originates. Day length changes consistently from year to year at a particular location and can provide a consistent and accurate signal th at the environment is favorable or unfavorable for reproduction. The magnitude of the change in day length between seasons decreases the closer one is to the equator (Vince-Prue, 1984). Thus, while photoperiod sensitive plants can be

PAGE 21

21 found at all latitudes, it is common to find da y-neutral plants near the equator (Wareing and Galston, 1963). In day-neutral plants, irradian ce and temperature, as well as the interaction of the two, are important factors that influence flowering (Bernier et al., 1993; Halevy, 1984). Light intensity and quantity appear to be involved in floral ini tiation and portions of floral development, while temperature is involved in the rate of floral development from initiati on to anthesis (Adams, 1999; Kinet and Sachs, 1984). Irradiance Influences Time to Flower Light quantity refers to the total am ount of light energy captured by a plant in a specified time period and is often quantified as th e daily light integral, or DLI (molm-2d-1). Increasing the light intensity will increase th e DLI and amount of photosynthetica lly active radiation available to the plant. Many day-neutral plants tend to ha ve a slight quantitative long-day response since increasing the daylength will increase the length of time per day that a plant can capture light energy (Kinet and Sachs, 1984; Korczynski et al., 2002). Across the United States, DLI varies from less than 5 molm-2d-1 in northern states in December to 55 to 60 molm-2d-1 in the Southwest in July and August (Korczynski et al., 2002). This wide range can greatly impact the time to flower for a crop and the length of time it occupies greenhouse space. The influence of light intensity and light quantity has been observed in a wide range of crops, and the general trend is that an increase in DLI will decrease the number of days to anthesis. In irradiance-responsive crops, the impact of an increase in irradiance will follow the law of diminishing retu rns. An identical increase in irradiance will have a much greater impact on reducing time to flower on plants grown under a low DLI compared to those grown under a higher DLI (Adams et al., 1997; Karlsson et al., 1989).

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22 One way to quantify the effect of irradiance on flowering is to observe the number of days to first flower in a species. Pelargonium x hortorum Radio grown at 22 Wm-2 reached 100% anthesis faster than those grown at 4 Wm-2 (Welander, 1983). Warner and Erwin (2005a) observed a decrease in the number of days to first flower in Antirrhinum majus Rocket Rose, Calendula officinalis Calypso Orange, Mimulus x hybridus Mystic Yellow, and Torenia fournieri Clown Burgundy as DLI increased. Piet sch et al. (1995) observed a decrease in number of days to flower in Catharanthus roseus Grape Cooler pl ants supplied with supplemental lighting. Fausey et al. (2005) re ported that an increase in DLI had a minimal impact on the time to flower in three perennials: Achillea and Gaura plants grown as 22 C flowered 5 and 7 d earlier, respectivel y, as DLI increased from 5 to 20 molm-2d-1 but no difference was observed in Lavandula A second way to quantify time to flower is to report the rate of progress towards flowering, defined as the reciprocal of the number of da ys to first flower (1/days to flower). Tagetes erecta Bonanza Yellow had an increased rate of progr ess towards flowering as irradiance increased from 5 to 25 molm-2d-1 (Moccaldi and Runkle, 2007), and chrysanthemum Resiliance had a higher rate of floral development under incr easing photosynthetic photon flux (PPF) levels (Warrington and Norton, 1991). Temperature Influences Time to Flower Floral developm ent is a metabolically-driven, te mperature-dependent process. From a base temperature to an optimal temperature, any incr ease in temperature will increase the rate of progress towards flowering. Above the optimal temperature, an increase in temperature will decrease the rate of progress towards flower ing (Armitage, 1994; Roberts and Summerfield, 1987). Karlsson and Werner (2001) observed that cy clamen grown at a cons tant temperature for eight weeks following formation of visible buds fl owered progressively earlier as temperatures

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23 increased from 8 to 20 C, but later as temperatur es increased from 20 to 24 C. The rate of progress towards flowering increased linea rly up to an optimum of 21.7 C in Viola x wittrockiana Universal Violet (Adams et al., 1997) and increased quadratically up to approximately 25 C in Celosia Gloria Mix (Pramuk and Runkle, 2005). With many crops, the specific co mbination of day and night te mperatures does not affect time to flower, but rather average daily temperatur e (ADT) is a better indi cator of the rate of floral development. The optimal ADT temper ature for flowering in chrysanthemum ranges between 17 and 22 C (Van de r Ploeg and Heuvelink, 2006). Campanula carpatica Blue Clips flowered approximately 20 d earli er as ADT increased from 15 to 25 C (Niu et al., 2001), Catharanthus roseus Grape Cooler flowered 30 d earlier between 18 and 35 C (Pietsch et al., 1995), Platycodon grandiflorus Astra Blue flowered 63 d earlier between 14 C and 29 C (Park et al., 1998), and Petunia xhybrida Wave Purple and Viola x wittrockiana Sorbet Blackberry Cream flowered 67 a nd 23 d earlier between 12 and 24 C, respectively (Mattson and Erwin, 2003). The rate of progress towards flowering in Salvia splendens Vista Red and Tagetes erecta Bonanza Yellow increased under higher temperatures (Moccaldi and Runkle, 2007). Temperature x Irradiance Interactions Influence Time to Flower Tem perature and light may interact to influen ce the time to flower. This interaction may hasten or delay flowering, or the effects of each other may be negated. In general, an increase in irradiance at lower temperatures is often more pronounced than at higher temperatures. In Petunia xhybrida Snow Cloud plants grown un der either 6.5 or 13 molm-2d-1, the higher DLI resulted in earlier flowering at all temperatures except at the two highest temperature treatments (25 and 30 C) (Kaczperski et al., 1991).

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24 In Salvia splendens Vista Red, as temperature increas ed from 15 to 25 C, the number of days to flower decreased from 42 to 24 d at 10 molm-2d-1 and from 37 to 21 d at 20 molm-2d-1 (Moccaldi and Runkle, 2007). In contrast, Impatiens wallerana Accent Red plants grown with less than 15 molm-2d-1 flowered earlier as temperature increased from 14 to 28 C, but no difference between temperatures was observed at the highest DLI treatments (Pramuk and Runkle, 2005). These differences may be attributed to the fact that salvia is a full-sun plant and impatiens prefer partial-shade locations in the la ndscape, and at the highest light treatments, the impatiens were exposed to supra-optimal DLI levels. Flowering in Angelonia Angelonia has been classified as a day-neut ral crop. In a study by Miller and Arm itage (2002), five AngelMist cultivars were grown under natural days, continuous short-days (SD), continuous long-days (LD), or various combinations of SD and LD (2 wk SD then LD, 4 wk SD then LD, 6 wk SD then LD, 2 wk LD then SD, 4 wk LD then SD, or 6 wk LD then SD), and the number of days to visible bud and anthesis we re not affected by photoperiod treatment. It a preliminary report, they noted th at all cultivars consistently fl owered between the 9th and 11th nodes (Armitage et al., 2000). Starman (2001) re ported that Blue Paci fic was also a dayneutral cultivar. These results are consistent with the fact that the native range for angelonia is tropical and subtropical regions. In the Caatinga (northeaster n Brazil, 36W 8S), where a number of field studies were conducted on flowering time and t ype of reproduction (Machado et al., 2006; Vogel and Machado, 1991), the natural phot operiod range is from 11.5 h in mid-June to 12.5 h in mid-December (U.S. Naval Observatory, 2008). Time to flower is angelonia is influenced by both temperature and irra diance. Angelonia is responsive to supplemental irradiance (Starman, 2001) and exhibits a quadratic response (Miller and Armitage, 2002). AngelMist Pink grown under na tural days or natural days plus either 900

PAGE 25

25 or 1200 molm-2s-1 continuous supplemental lighting from high intensity discharge (HID) lights flowered in 57, 49, and 45 d, respectively. In production, recommended light levels range between 6,000 to 10,000 foot candles (approximately 1200 to 2000 molm-2s-1) (Ball FloraPlant, 2006). An increase in temperatur e, up to an optimum, hastens flowering. AngelMist Pink grown at consta nt temperatures of 15, 22 or 30 C flowered in 58, 47, and 50 d, respectively, and exhibited a quadratic response (M iller and Armitage, 2002). This would indicate that the optimal temperature for flower ing is between 22 and 30 C. It is unknown if other cultivars have similar responses to temperature as AngelMist Pink. In production, the recommended growing temperatures are 17 to 20 C night and 25 to 30 C day (Ball FloraPlant, 2006). No studies have been publ ished to date on the combined influence of temperature and irradiance on time to flower in angelonia. Factors Influencing Continued Flowering In m ost warm-season crops, temperatures above 30 C will result in a decrease in flower number (Armitage, 1994), but this threshold valu e will vary depending upon genus, species, and even cultivar. Also, the length of the high temperature stress will influence the degree of response observed. This decline in flowering has been attributed to a carbon shortage within the plant. At higher temperatures, especially high night temp eratures, plants will have a higher rate of photorespiration and a lower daily net photosynthesis. Flowers and developing inflorescences will compete with other active sinks in the plant for a limited carbon supply, and over time, fewer flowers will be able to develop. High temperature stress on flowering has also been shown to influence signaling within the floral pathways. In Arabidopsis thaliana floral abortion occurred when whole plants or inflorescences onl y were subjected to temperatures above 33 C (Warner and Erwin, 2005b). Based on a degree-hour s (C-h) model, flower buds began aborting

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26 between 200 and 300 C-h and complete inflorescences aborted above 300 C-h. Thus, while less carbon assimilation at high temp eratures influences flowering in many crops, there are most likely other crops in which the inflorescence dete cts changes in temperature and translocates signals within the plant leading to floral abortion. Most of the published resear ch on the influence of high te mperatures on flowering has been conducted on vegetable and field crops, in which a reduction in flowering or fruit set results in decreased yield. In three field-grown summer brassica (canola) species ( Brassica napus B. rapa, and B. juncea ), an increase in temperature resu lted in lower flower number, and temperatures above 29.5 C resulted in significan t yield loss (Morrison an d Stewart, 2002). In pepper (Capsicum annuum ), a comparison of plants grown at 25 and 33 C indicated that the number of flower buds and mature flowers was not affected by the increase in temperature, but fruit set was less at 33 C (Erickson and Markhart 2001). This would indicate that while flowers developed, there was a subsequent problem with pollen development, ovary development, and/or fertilization. In tomato ( Lycopersicon esculentum ), chronic high temperat ures influence pollen grain development and leads to re duced fruit set (Sato et al., 2000). Studies looking at the impact of temperatur e on flower number or flower bud abortion in greenhouse crops have been limited to time of first flower. This is of importance because decreased flowering will impact plant quality an d crop marketability. Ka czperski et al. (1991) attributed a delay in flowering in Petunia xhybrida at temperatures above 25 C to floral bud abortion. In a study conducted by Warner and Er win (2005a) on five ornamental plant species, the number of flower buds presen t at time of first flower was less for plants grown at 32 C relative to 20 C. Impatiens wallerana was the least sensitive, w ith a 30 percent decrease in flower bud number, and Torenia fournieri was the most sensitive, with a 95 percent decrease. In

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27 a screen of 12 Viola x wittrockiana cultivars, the number of flower buds present at time of first flower was 20 to 77 percent less in those plants grown at 30 rather than 20 C (Warner and Erwin, 2006). Angelonia cultivars were observed to stop flowering or have significantly fewer inflorescences per plant in the University of Florida trial garden dur ing the summer 2005 and 2006 seasons (Boldt, personal observation). This reduction in flowering has not been observed in northern states, where it is pl anted in early summer and flow ering is continuous until first frost. It is unknown if the reduction in or cessation of flowering in angelonia cultivars is due to chronic high temperature stress or advanced plan t age since it is planted outside much earlier in the year in the southern states relative to northern states. In a study by M iller et al. (2001), net photosynthesis in two angelonia cu ltivars was optimal at 20.8 for AngelMist Purple Stripe and 19.8 C for AngelMist Deep Plum. In locatio ns where both the day a nd night temperatures exceed this range during the summer months, it coul d be expected that flowering may be reduced due to lower net carbon assimilation. Plant Growth Temperature Tem perature has an impact on plant growth, flowering, and plant quality. It is known to affect plant height, number of late ral shoots, flowering time, flower number, and flower size. In many warm season crops, temperatures below 10 C will significantly slow plant growth and temperatures above 30 C will cause a reduction in plant quality (Armitage, 1994). Temperature impacts plant height by altering th e number of nodes produced in a specified time period and by altering internode length (Warner and Erwin, 2001b). As temperature increases within the optimal range, the leaf unfolding rate will increas e and number of nodes produced by a plant will increase. This process is dependent upon average daily temperature, not

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28 day or night temperatures, and it will reach a plateau at approximately 24.4 to 26.6 C (76 to 80 F). Internode length in plants is affected not by average day and night temperatures, but rather the magnitude of the difference between them. This is termed DIF, and the greater the DIF (higher day relative to night temp erature), the greater the inter node length (Erwin et al., 1989). A study by Miller and Armitage (2002) reported that angelonia plant quality was greatest at 22 C when plants were grown at a constant 15, 22, or 30 C. Plants grown at 15 C were chlorotic and stunted, while plants grown at 30 C we re tall with thin, brittle stems. Plant height was 31, 52, and 68 cm, respectively, and responded linea rly to temperature. This is similar to results observed in Petunia xhybrida Snow Cloud by Kaczperski et al. (1991), in which plant height increased linearly and in ternode length increased quadratically as temperature increased. In addition to plant height, temperature can affect flower size a nd flower number. A decrease in flower size in response to in creased temperatures has been observed in Chrysanthemum morifolium (Willits and Bailey, 2000); Calendula officinalis Impatiens wallerana Mimulus x hybridus and Torenia fournieri (Warner and Erwin, 2005a); and Viola x wittrockiana (Warner and Erwin, 2006). The number of flowers per plant has been shown to decrease in Viola x wittrockiana and Petunia x hybrida, increase in Impatiens wallerana and not change in Viola x wittrockiana in response to increased temp eratures (Mattson and Erwin, 2003; Warner and Erwin, 2006). Irradiance Increased irradiance is g enerally associated with an increase in plant quality due to improved plant architecture and an increase in fl ower size. Fausey et al. (2005) observed an increase in lateral branching of Achillea Gaura and Lavandula as DLI increased from 5 to 20 molm-2d-1 and a two to three-fold increase in the number of inflorescences on Achillea and Gaura A linear increase in flower size was reported for cyclamen plants grown at 20 C as DLI

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29 increased from 1.8 to 21.6 molm-2d-1 (Karlsson et al., 1989). An increase in flower diameter has been reported for Calendula officinalis Impatiens wallerana Mimulus x hybridus and Torenia fournieri when grown at 21.8 versus 10.5 molm-2d-1 (Warner and Erwin, 2005). Supplemental lighting or a na tural increase in irradiance ha s been shown to increase angelonia plant quality. Blue Pacific plants grown under HID supplemental lighting were more compact and better branched (Starman, 2001). Holcombe et al. (2001) have reported that angelonia plants grown under 6 molm-2d-1 had very weak lateral branches, plants grown at 12 molm-2d-1 were of acceptable quality, but 18 molm-2d-1 was necessary for an adequate number of inflorescences. Temperature x Irradiance Interactions The interaction of tem peratur e and light can have a major impact on plant quality. In Gypsophila paniculata L., the temperature corresponding to the highest plant quality decreased under higher irradiance levels. At a light level of 450 molm-2s-1, the temperature optimum was 20 C, but at 710 molm-2s-1, the temperature optimum was between 12 and 20 C (Hinkleton et al., 1993). In Salvia splendens Vista Red, however, the temperature corresponding to maximum plant height increas ed as irradiance increased. At 5 molm-2d-1, plant height was greatest at 20 C, while at 25 molm-2d-1, plant height was greatest at 24 C (Moccaldi and Pramuk, 2007). It is unknown whether or not this increase in plant height was a desirable or undesirable char acteristic for this crop. In angelonia, no research has been publishe d on whether temperatur e and light have an interactive effect on plant height or overall plant quality. Ange lonia plants grown under natural days in the spring and summer months will be expos ed to increasing temperature and light levels, but by varying amounts.

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30 Fertilizers Plant Growth Use of the correct fertilizer and concentr ation during crop produc tion can significantly im pact final quality of the finished product. Pl ant height, plant size or fullness, leaf color, number of flowers or inflorescences, time to flow er, and dry weight can be impacted by the type and concentration of fertilizer applied. Growth parameters, such as height and dry weight, tend to increase in response to increasing fertilizati on up to an optimum, and then decrease (James and van Iersal, 2001). Flowering has been shown to be delayed at low nutrie nt concentrations in Salvia splendens (Kang and van Iersal, 2004). The optimal fertilizer concentration to use in production varies by crop, cultivar, irrigation method, frequency of applicati on, current nutritional status of the crop, and environmental conditions during crop production. It has been s hown that the light leve l during production does not influence the optimal fe rtilizer concentration for Begonia semperflorens and Petunia xhybrida examples of low-fertility and high-ferti lity requiring crops (Nemali and van Iersal, 2004), but temperature influences optimal fertili zation in petunia (Kang and van Iersal, 2001). As temperature increased, the optimal concentr ation necessary for plant growth decreased. Fertilizer recommendations vary dependi ng upon irrigation method. Leaching does not occur in a subirrigation system and the soluble salts applied are either used by the plant or accumulated in the potting medium. However, le aching can occur with overhead irrigation and excess salts may be leached as needed to mainta in the correct electrical conductivity (EC) range for optimal plant growth. It has been recommende d that subirrigated crops are fertilized with a half-strength solution relative to the suggested concentrati on for overhead watering (Nelson, 1994). Klock-Moore and Broschat (2001) observed that subirrigated petu nias at fertilizer

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31 concentrations of 50, 100, or 150 mgL-1 had final EC levels that we re twice as high as overhead irrigated plants at 50 and 100 mgL-1 and five times as high at 150 mgL-1. Crops vary in their sensitiv ity to fertilization. Optimal growth has been observed to range from N at approximately 100 mgL-1 for Celosia argentea Impatiens xhawkeri Impatiens wallerana and Zinnia elegans (Kang and van Iersal, 2002; Kent and Reed, 1996; Whipker et al., 1999) to up to 400 mgL-1 in Gomphrena globosa and Matthiola incana (Kang and van Iersal, 2002). Different plant quality parameters may have different optimal concentrations and a grower will need to determine which factor is of most importance. For instance, Petunia xhybrida dry weight was maximized when supplied with a complete fertilizer with N at 355 mgL-1, height at 255 mgL-1, and flower number at 165 mgL-1 (James and van Iersal, 2001). Within a crop, cultivars may have different fertilizer optimums. Double impatiens ( Impatiens wallerana ) Purple Magic, a green-leaved cultivar required a higher concentration than Blackberry Ice, a variegated-leav ed cultivar (Whipker et al., 1999). Cultivar differences have been noted in poinsettia ( Euphorbia pulcherimma ) related with leaf color. Dark-green leaved cultivars require an electrical c onductivity (EC) of 1.5 to 2.0 mmhoscm-1 (mScm-1) using the saturated media extract (SME) procedure, wherea s medium-green leaved cultivars require 2.0 to 2.5 mmhoscm-1 (Ecke et al., 2004). Use of a Hand-held Meter to Measure Chlorophyll Content The SPAD m eter (Minolta Co., Osaka, Japan) is an instrument used to measure leaf chlorophyll content of a crop quick ly and non-destructively. Out put values do not have a unit associated with them, but they can be correlate d with leaf chlorophyll co ntent, with increasing values indicating higher chlorophyll levels. The nature of the rela tionship varies. It has been reported to be linear for ornamental foliage plants (Wang et al., 2005); non-linear for Oryza sativa Glycine max, Triticum aestivum and Solanum tuberosum (Monje and Bugbee, 1992;

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32 Uddling et al., 2007); and exponential for Glycine max and Zea mays (Markwell et al., 1995). SPAD values have been shown to correlate strong ly with the actua l chlorophyll content of leaves as determined through destructive sampling (r2 values of approximately 0.9 or greater) (Monje and Bugbee, 1992; Uddling et al., 2007; Wang et al., 2005). SPAD meters were initially used in field crops to determine if supplemental N was necessary for optimal yield. Nitrogen deficienci es show up as lighter green foliage coloration due to a decrease in the chlor ophyll content of the l eaf. Greenhouse and bedding crops grown in soilless media have their fertilizer requirements met through the application of a water-soluble or slow-release fertilizer. These tend to be comple te fertilizers, but their concentrations are reported as N concentration in mgL-1 or ppm. Increasing the fertilizer N concentration applied will result in an identical increase in concentrat ion of all other elements present and an increase in the total concentration of so luble salts available to the crop In angelonia, when single elements were withheld from an otherwise comp lete fertilizer, deficiency symptoms for N, P, and K all appeared at the same number of days after start of treatment (Williams, 2004). Thus, the use of SPAD meter can indicate that plants with lower values will likely have lower tissue concentrations of other elements as well. Cultivar and environmental factors can influence SPAD readings, including normal leaf coloration under adequate N fertilization, relative water status of the plant, temperature, and sunlight (Peterson et al., 1993). Leaf size should not result in significan t variation between samples since the measurement area for the meter is very small, 2 mm by 3 mm (Minolta, 1989). Leaf thickness ranging from 0.19 to 0.66 mm did not affect SPAD readings (Wang et al., 2005) and Minolta states that the meter can determine a value for leaves up to 1.2 mm thick (Minolta, 1989). Markwell et al. (1995) re ported that the exponential equa tion developed for soybeans and

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33 corn fit data collected for sorghum and Arabidopsis and suggested that multiple crops may have very similar correlations between the SPAD valu e and actual chlorophyll content. Wang et al. (2005), however, reported that linea r models developed for 10 diffe rent foliage crops were not identical and that individual mode ls should be developed for use across crops, but may be useful for comparing cultivars. Published fertilizer guidelines for angeloni a vary by company. All recommend constant liquid feeding with a complete fertilizer, but the level of fertilization ra nges from N at 150 to 250 mgL-1 (Ball FloraPlant, 2006; Fisc her, 2008; PanAmerican Seed, 2006; Proven Winners, 2006). Fertilizer guidelines published in trade press articles r ecommend N at concentrations ranging from 75 to 200 mgL-1 (Armitage, 1997; Schoellhorn and Al varez, 2002; Smith, 2007). It is unknown whether these differences in recommende d levels are due to cultivar, series, or environmental conditions. Plant Growth Regulators Introduction Plant growth regulators (PGRs) ar e used in the floriculture i ndustry to control plant growth during production, im prove quality, and increase longevity at retail (A rteca, 1996). These chemicals inhibit stem elongation without killing the meristem (Cathey, 1964), and plants treated with a PGR look identical to untrea ted plants except for the fact that they are shorter. The first PGRs were introduced in the 1960s, and since then two major groups have arisen based on mode of action. One group inhibits stem elongation by inhibiting various steps in the gibberellin synthesis pathway and this includes daminozide and paclobutrazol. The second group releases ethylene within the plant, resulting in an inhib ition of stem elongation and an increase in lateral branching in certain crops, and this group includes the chemical ethephon (Rademacher, 2000).

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34 The effect that a PGR will have on a plant is dependent upon multiple factors, including species sensitivity, cultivar sensitivity, plant ag e, PGR concentration, method of application, and environmental conditions under which the plant is grown (Barrett, 2001; Fletcher et al., 2000; Rademacher, 2000). Crops may exhibit sensitivity to some, all, or none of the PGRs typically used in production. For example, p aclobutrazol inhibited stem elongation in Hibiscus coccineus H. radiatus and H. trionum but daminozide was effective only on H. trionum (Warner and Erwin, 2003). In a trial of 26 ornamental cabbage and kale cultivars conducted by Gibson and Whipker (2001a), only two cultivar s were responsive at 2500 mgL-1 and eight at 5000 mgL-1. Plant age can impact responsiveness to a PG R application, especially for foliar-applied chemicals, since more chemical is taken up by younger leaves due to the presence of a thinner cuticle (Sachs and Hackett, 1972). The environmental conditions in which the crop is grown can greatly a ffect efficacy. In warmer temperatures, plants are growing at a faster rate than those grown at cooler temperatures. They usually will require the application of a higher concentration or multiple applications in order to see a similar amount of height control relative to thos e grown at cooler temperatures (Barrett, 2001). Growers typically adjust the concentr ation of a PGR appli cation between winter and summer applications. The following sections will describe in greater de tail each of the three PGRs to be used in this research: paclobutrazol, daminozide, and ethephon. Reported similarities and differences among crops and between cultivars will be discusse d, as well as any known results pertaining to angelonia cultivars. Paclobutrazol Paclobutrazol [(2RS,3RS)-1-(4-chlorophe nyl)-4,4-dim ethyl-2-(1H-1,2,4-triazol-1yl)pentan-3-ol], discovered in the 1960s during tr ialing for new fungicides (Fletcher et al., 2000),

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35 is a member of the triazole family. Trade names for paclobutrazol in clude Bonzi, Downsize, Paczol, Piccolo, and PPP-333. It inhibits gibberell in (GA) synthesis by inhibiting the function of cytochrome P450 and affecting the formation of ent -kaurenoic acid from ent -kaurene, GA precursors (Fletcher, 2000; Rademacher, 2000). Treated plants will exhi bit shorter internodes and thicker, darker green leaves. In chrysant hemum, this increased leaf thickness has been reported to be due to the formation of an additi onal layer of palisade ce lls (Burrows et al., 1992). Paclobutrazol, effective as a media drench or foliar spray, is translocated through the xylem only (Barrett, 2001). The application of a foliar spray will be effective at inhibiting internode elongation only if it comes in contact with stem tissue and developing meristems, so good spray coverage is essential (Barrett, 2001). Media drenches are highly effective and very low concentrations are needed compared to foliar sprays. Paclobutrazol is not very water-soluble and will rapidly move into the wax layers present in the leaf. In a study by Chamel et al. (1991), paclobutrazol uptake by leaves was determined but no distinction was made as to whether it was adsorbed and/or absorbed, thus they referred it as soprtion. So rption was rapid during the first hour following application and almost all was sorb ed within 24 h of appl ication. These results concur with findings from Barrett et al. (1987) in which the efficacy of a spray application was not reduced when plants were ove rhead irrigated as soon as 30 mi nutes after app lication. While paclobutrazol rapidly moves out of solution to the cuticle, Chamel et al. (1991) determined that only a very small amount, approx imately 5 percent, penetrates the cuticle and into the leaf. The timing of an application will have an im pact on the amount of efficacy observed. When Chrysanthemum Bright Golden Anne plants were sprayed at 0, 2, or 4 weeks following pinch, the earlier applications resulted in incr eased efficacy and shorter plants at flowering relative to the later applications and untreated plants (Gilbertz, 1992). This increased efficacy

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36 early in the crop may be due to a combination of factors, but most likely is due to the reduced canopy size at the time of application. When applied at pinch, it is easier to get good spray contact with the stem and more soil media will be sprayed, resulting in a slight drench effect. Recommended concentrations for crops range between 5 and 90 mgL-1 for foliar sprays and 1 mgL-1 for media drenches (Syngenta, 2005) a nd may be adjusted depending upon species or cultivar sensitivity. The triazoles are very active and usually only very low concentrations are necessary for efficacy (Barrett, 2001). Paclobutra zol is less active than uniconazole [(E)(S)-1(4chlorophenyl)4,4-dimethyl-2(1,2,4-tri azol-1-yl)pent-1-ene-3-01] and two to four times as much is needed for similar efficacy, but it is much more active than daminozide. In Scaevola aemula a media drench of 1 mgL-1 uniconazole had similar efficacy as a 4 mgL-1 paclobutrazol drench (Starman and Williams, 2000). Paclobutrazol and uniconazole sprays at 30 and 15 mgL-1, respectively, provided si milar height control in Chrysanthemum Bright Golden Anne (Gilbertz, 1992). Cultivar sensitivity has been reported in ornamental cabbage and kale, with Osaka White exhibiting a linear response and Nagoya Red exhibiting a linear-plateau response to increasing concentrations of p aclobutrazol (Gibson and Whipker, 2001b). This indicates that Osaka White is less sensitive than Nagoya Red. Potential differences in cultivar sensitivity have been reported in angelonia. A spray concentration of 50 mgL-1 was effective for AngelMist Pink (Mil ler and Armitage, 2002), but concentrations up to 80 mgL-1 were ineffective for Blue Pacific (Starman, 2001) However, it is not known whether these differences can be explained by cultivar sensitiv ity or differences in environmental conditions during production. No recommendations for foli ar sprays have been published in cultural guidelines or the trade press, but drench recommendations vary from 1 to 16 mgL-1 (Armitage,

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37 1997; PanAmerican Seed, 2006; Schoellhorn and Alvarez, 2002). Proven Winners (2006) recommends the application of 2 to 5 mgL-1 uniconazole on Angelface Blue and 5 to 10 mgL-1 on Angelface White and Angelface Blue Bicolor, indicating that differences in cultivar sensitivity likely exist in this series. Howeve r, it is not known if other cultivars differ within series and/or across series. Daminozide Da minozide [butanedioic acid mono (2,2-dimet hylhydrazide)] was first described in 1962 and is known by the trade name B-Nine (Pla nt Growth Regulator Working Group, 1977). Treated plants exhibit shorter internodes and are sm aller than untreated plants due to its effect on inhibiting the synthesis of GA (R ademacher, 2000). It is much le ss active than th e triazoles and in responsive species, concen trations of 2500 to 5000 mgL-1 are recommended (Chemtura, 2001). Daminozide is applied as a spray and is ve ry mobile within the plant (Barrett, 2001). Unlike paclobutrazol, it is very water soluble an d moves slowly from the aqueous solution to the cuticle. It can be washed off if leaves are watere d within 4 h following applic ation (Barrett et al., 1987). Due to the high concentration needed fo r efficacy, daminozide is more effective when used in winter months or combined in a tank mix with chlormequat ch loride [chlormequat (2chloroethyl)trimethylammonium chlori de; Cycocel] (Barrett 2001; Cathey, 1975). Plant response to daminozide varies with crop and cultivar. In a study by Gibson and Whipker (2000a) on 26 ornamental cabbage and ka le cultivars, two were responsive at 2500 and eight at 5000 mgL-1. Lewis et al. (2004) observed a cul tivar differences in pansy in which Colossus Yellow Blotch and Delta Pure Yellow were responsive at 4500 mgL-1 but Majestic Giants Purple was unresponsive at all concentrations. In contrast, osteospermum ( Osteospermum ecklonis ) cultivars Congo and Wildside both were insensitive at concentrations of up to 10,000 mgL-1 (Gibson and Whipker, 2003).

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38 Angelonia AngelMist Pink has shown a response to daminozide at 2500 and 5000 mgL-1 and Blue Pacific at 5000 mgL-1 (Miller and Armitage, 2002; Starman, 2001). Selecta First Class (2008) recommends daminozide at 2500 mgL-1, and PanAmerican Seed (2006) and Ball FloraPlant (2006) recommend tank mixes of daminozide and chlormequat chloride for height control. Ethephon Ethephon (2-chlorophenylphosphonic acid) was first synthesized in 1946, but its m echanism of decomposition was not reporte d until 1963 (Beaudry and Kays, 1988). Under high pH conditions, it naturally breaks down to form chlorine, phosphonic acid, and ethylene (Beaudry and Kays, 1988). The release of ethylen e is the key by-product of this reaction which affects plant growth. Ethylene, a simple hydroc arbon, is the smallest known plant hormone in the plant and is involved in a myriad of plant processes thro ughout its life cycle, from seed germination to plant senescence (Arteca, 1996). Ethephon is consid ered a plant growth regulator because the release of ethylene in the plant can result in an inhibition of stem elongation, increased lateral branching, and/or delayed flower ing. It is marketed under the trade names of Florel, Pistill, and Ethrel and applied at concentrations ranging from 250 to 1000 mgL-1 (Barrett, 2001). Ethylene is known to inhibit cell elongation and interact wi th auxins to affect the outgrowth of lateral buds. Cells treated with ethylene will have a similar total volume relative to untreated cells but their elongati on is inhibited (Osborne, 1974). Burg (1973) reported that this is due to a reorientation of the microfibrils in the cell wall such that l ongitudinal growth but not lateral growth is restricted, expl aining why ethylene-treated plants may have thicker stems. The mechanism underlying the release of lateral buds from apical dominance and their subsequent growth is believed to be due to a shift in hormone rati os within the plant. In Petunia x hybrida

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39 the application of ethephon to plants resulted in a 20 percent decrease in IAA (indole-3-acetic acid) and a 24 percent decrease in the auxin:cy tokinin ratio relative to untreated plants; no changes were observed in cytoki nins between treated and untreat ed plants (Haver and Schuch, 2001). The efficacy of an ethephon application depe nds upon the plant and the conditions at the time of application, including solution pH, te mperature, relative humidity, plant species and cultivar. Ethephon is stable below pH 5.0 but w ill begin to decompose and release ethylene in higher pH environments (Warner and Leopold, 1969), with 98 percent decomposition seen in an ethephon solution maintained at pH 7.4 at 25 C (Segall et al., 1991). In a study examining the rate of decomposition of ethephon, it increased significantly between pH 6.0 and 8.0 at 25 C. However, temperature greatly influenced the magnitude of the decompos ition, with similar rates observed between a solution with pH 9.1 kept at 30 C and a solution with a pH of 6.1 kept at 40 C (Biddle et al., 1976). Differences between spec ies and cultivars may be due to differences in ethylene sensitivity or due to differences in the leaf morphology (Beaudry and Kays, 1988). In leaves with a thinner cuticle, pe netration of the chemical is gr eater than those with a thicker cuticle. Ethephon or ethylene can be translocated th rough the plant. Puech and Crane (1975) observed the presence of 14C in the translocation stream with in 2 h of an ethephon application, but it was not reported if it was et hephon or the released ethylene that was being translocated. Kwong and Lagerstedt (1977) obs erved the accumulation of 14C in apical meristems and nodes 1 h following application, but again, no distincti on was made as to whether it was ethylene or ethephon. In chrysanthemum, the application of a single drop of ethephon to a leaf was significant enough to affect stem elongation and delay flowering, and this effect was still

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40 observed if the treated leaves were remove d 12 h following applica tion, indicating that translocation or signaling occurr ed within that time interval (Stanley and Cockshull, 1989). Common responses to ethephon applications include a reduction in stem elongation, an increase in lateral branching, and delayed flower ing. Decreased plant he ight has been observed in Achillea Echinacea Leucanthemum Monarda, and Phytostegia (Hayashi et al., 2001). Foley and Keever (1992) and Carpenter and Carlson ( 1972) observed an increase in the number of laterals in geranium Hollywood Star and D ark Red Irene plants treated with 500 and 1000 mgL-1, respectively. Glady et al. (2007) reported increased branching of Coreopsis verticillata Moonbeam, Dianthus caryophyllus Cinnamon Red Hots, and Veronica longifolia Sunny Blue Border following ethephon applications. De layed flowering of more than 2 weeks has been reported for ethephon-treated plants. Scaevola treated with ethephon at 500 and 1000 mgL-1 flowered 8 and 11 d later than untreated plants, respectively (Starman and Williams, 2000), and geranium Dark Red Irene treated with 1000 mgL-1 flowered 16 d later (Carpenter and Carlson, 1972). One drawback to the use of ethephon is the appearance of phyto toxicity symptoms following application in some, but not all crops. Hayashi et al. (2001) trialed eight species of perennials and only Monarda exhibited leaf necrosis. Gla dy et al. (2007) observed deformed growth and leaf necrosis in Dianthus caryophyllus Cinnamon Red Hots, but not in the two other crops also in the experiment. Cultivar differences have been reported following ethephon sprays. Hammond et al. (2007) observed more compact growth and delayed flowering in Gaillardia pulchella Torch Flame but no difference in plant growth between treated and untreat ed plants of a native Florida ecotype. In a study of 23 cultiv ars from 12 different genera, Fa ust and Lewis (2005) indicated

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41 that crops differed in response, but all cultivars within a particular genus responded similarly. In contrast, Starman et al. (2004), trialed 27 cult ivars across 16 genera and observed cultivar interactions for plant height, pl ant width, and number of days to flower. For example, treated plants were significantly shorter than untreated plants for all cultivars of Diascia xhybrida Impatiens wallerana Lantana camara and Nemesia xhybrida In Antirrhinum majus Calibrachoa hybrid and Petunia xhybrida, certain cultivars were re sponsive and others were nonresponsive. Ethephon has been reported to be an effec tive PGR for angelonia Blue Pacific and no phytotoxicity symptoms were reported (Starm an, 2001). Proven Winners (2006) and Ball FloraPlant (2006), however, recommend against the use of ethephon due to its potential to cause leaf tip burn and distorted growth, but this ha s not been mentioned in the production guidelines of other companies. Objectives 1. Conduct a cultiv ar screen of commercial Angelonia angustifolia cultivars and determine the range of variability present within this species for growth and flowering traits. Of interest will be how variable the cultivar s are within each series and across series. From this set of cultivars, a sub-sample of cultivars representing this range of variability will be selected for use in subsequent experiments. 2. Determine the effect of season on time to flower and plant growth on se lect cultivars grown under natural days in the greenhouse during the main production window (late winter through early summer). 3. Determine the effect of three plant dates on the summer landscape performance of select cultivars. 4. Determine the effect of increasing fertilizer c oncentrations on time to flower, plant growth, and flowering characteris tics of select cultivars grown in the greenhouse. 5. Determine the effect of paclobutrazol, da minozide, and ethephon on growth of select cultivars during greenhouse production and identi fy if they differ in their response to chemical and/or concentration applied

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42 Table 1-1. Current taxo nomic classification of Angelonia angustifolia Kingdom Planta Division Magnoliophyta (Angiosperms) Class Magnoliopsida (Dicots) Subclass Asteridae Order Scrophulariales Family Plantaginaceae Genus Angelonia Humb. & Bonpl. Table 1-2. Commercially av ailable angelonia series. Series Company Propagation method Alonia Danziger Vegetative cuttings Angelface Proven Winners Vegetative cuttings Angelina Selecta First Class Vegetative cuttings AngelMist Ball Horticultural Company Vegetative cuttings Carita Syngenta Flowers Vegetative cuttings Serena Ball Horticultural Company Seed

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43 Table 1-3. Cultivar names of angelonia used in experiments. Cultivar Statusz Plant code Plant patent Angelface Blue Commercially available ANBLAUZWEI PP 14,189 Angelface Blue Bicolor ? Angelface Dresden Blue Commercially available ANSKY PP 17,155 Angelface Pink Commercially available ANPINK PP 16,818 Angelface Wedgewood Blue Commercially available ANWEDG PPAF Angelface White Discontinued ANWHIT PP 13,179 Angelina Blue Commercially available Angelina Dark Blue Commercially available Angelina Pink Commercially available Angelina Pink and White ? Angelina Violet and White Commercially available Angelina White Commercially available AngelMist Basket Pink AngelMist Spreading Pink BALANGBAKIN PPAF AngelMist Basket Purple AngelMist Spreading Purple BALANGBEKE PP 15,546 AngelMist Basket White AngelMist Spreading White BALANGSPRI PPAF AngelMist Dark Lavender AngelMist Purple BALANGDARLA PP 17,516 AngelMist Dark Pink Commercia lly available BALANGDARPI PP 17,071 AngelMist Dark Rose Commercially available BALANGDAROS PP 17,243 AngelMist Deep Plum Imp. ? AngelMist Lavender Commercially available BALANGLADER PP 17,909 AngelMist Lavender Stripe Commercially available BALANGLAST PP 16,668 AngelMist Pink Commercially available BALANGPIKIM PP 17,515 AngelMist Plum Commercially available BALANGPLUM PP 17,232 AngelMist Purple Improved Discontinued BALANGIMPU PP 13,921 AngelMist Purple Stripe Commer cially available BALANGPRIPE AngelMist White Cloud ? AngelMist White Improved Commercia lly available BALANGWITIM PP 16,501 Serena Lavender Commercially available Noney Serena Lavender Pink Commercially available Noney Serena Purple Commercially available Noney Serena White Commercially available Noney z Commercially available, discontinued, or renamed. y Seed propagated cultivar.

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44 CHAPTER 2 Angelonia angustifolia CULTIVAR SCREE N Introduction Angelonia, a genus native to Central and South Am erica (Huxley, 1992), contains approximately 30 species and Angelonia angustifolia is the only species grown commercially. Iinitially it was commercially ava ilable as non-patented cultivar s, but its popularity was limited due to the incidence of viruses. In the la te 1990s, the first virus-indexed cultivars were introduced and this crop saw a resurgence in popularity. Since then, multiple companies have developed angelonia breeding programs and have re leased cultivars with a range of growth and flowering characteristics and landscape performance. The objective of the first experiment was to conduct a cultivar screen on 31 commercially available cultivars from three companies and qua ntify the range of phenotypic variability present in growth habit, time to first flower, flower size, and powdery mildew susceptibility. The second experiment was designed to look at differences in flowering and gr owth habit in a small set of cultivars over multiple production seasons. Th ese cultivars were selected based upon an observational cultivar screen conducted in summ er 2006 and were selected for their range of flowering and growth habits. Materials and Methods Experiment 2-1 Cultivar Screen of 31 Commercial Cultivars Rooted cuttings or plugs (seedli ngs) of 31 commercially available Angelonia angustifolia cultivars were received the s econd and third week of January 2007 (Ball FloraPlant, West Chicago, IL; Costa Farms, Miam i, FL; EuroAmer ican Propagators, Bonsall, CA; Sunshine Greenhouses, Provo, UT). Nine plants of each cultivar were planted on 25 Jan. into

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45 2.2 L (16 cm diameter) pots filled with Fafard 2 soilless medium (Conrad Fafard, Inc., Apopka, FL) and pinched to three nodes on 30 Jan. Plants were watered as needed and continuously liquid fed with N at 150 mgL-1 using a 20.0N-4.4P-16.6K fertilizer (Peters 20-10-20 Florida Special, The Scotts Co., Marysville, OH). The experimental design was a complete randomized block design, with three blocks and three plants of each cultivar per block. Over the length of the experiment the average daily minimum, ma ximum, and 24-h temperatures were 17.0, 28.0, and 21.4 C, respectively. The average daily light level was 20.2 molm-2day-1. Plants were observed daily and the time from planting to first open flower and marketability were recorded. An open flower was defined as having th e corolla (both upper and lower lips) unfurled. Marketability was defined as when a plant had at least 20 open flowers, regardless of the number of inflorescences w ith open flowers. When each plant reached marketability, plant height and width were measured and the number of inflorescences with open flowers was determined. Plant height was measured from the top of the po t to the tallest growing tip. Plant width was measured at the widest point in the plant canopy and then again perpendicular to the first measurement. Average plant width was used for analysis. To provide an additional comparison of the characteristics for the cultivars at the sa me point in time, on 6 Apr. (9.5 weeks after pinch), the number of infl orescences with at least one open flower was counted, flower height and width was measured with caliper s, and plants were evaluated for powdery mildew susceptibility. Powdery mildew susceptibility was rated on a 1-5 scale with 1 = no powdery mildew present, 2 = slight presence of powdery mildew, 3 = moderate presence of powdery mildew, 4 = severe presence of powdery mildew, and 5 = leaves completely covered with powdery mildew.

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46 Data were analyzed using SAS 9.1 (SAS Institu te, Cary, NC) for analysis of variance and mean separation using Waller-Duncan at =0.05. Experiment 2-2 Cultivar Comparison of Seven Cultivars across Multiple Seasons Seven Angelonia angustifolia cultiva rs were selected from a preliminary screen of 25 cultivars conducted in July a nd August 2006 in order to encomp ass a wide range of growth habits, flower size, and time to flower (Table 2-1). These cultivars are the following: Angelina Violet and White, AngelMist Dark Lavender, AngelMist Lavender Stripe, AngelMist Purple Improved, AngelMist Purple Stripe, Angelface Blue, and Angelface White. Each cultivar was propagated from unrooted cuttings and grown to finished size at four different seasons: late winter, early spri ng, late spring, and early summer. Plants for the late winter experiment were th e same as those used in Expt. 2-1. For the other three seasons, stock plants were grown in 2.2 L (16 cm diamet er) pots at the University of Florida (Gainesville, FL) on a subirrigation bench and continuously liquid fed with a 20.0N4.4P-16.6K fertilizer (Peters 20-10-20 Florida Sp ecial, The Scotts Co., Ma rysville, OH) with N at 75 mgL-1. Stock plants were replaced every three months with new virus-indexed plants obtained from the breeder companies in order to maintain clean stock. Cuttings of each cultivar were harvested and stuck in 25 mm moistened Ellepots (Knox Nursery, Winter Garden, Florida) on the following dates: 19 Feb., 28 Mar., and 9 May 2007. They were placed under intermittent mist for 2 to 3 weeks and acclimatized in a natural ventilated greenhouse for 1 week. During acclimatization, rooted liners were watered as needed and continuously liquid fed with N at 150 mgL-1 using 20.0N-4.4P-16.6K. Rooted liners (one plant per pot) were planted on 25 Jan., 14 Mar., 19 Apr., and 31 May into 2.2 L pots filled w ith Fafard 2 soilless medium (Conrad Fafard, Inc., Apopka, FL) and pinched to three nodes 5 to 7 days later. Day 0 for each of the four seasons was 30 Jan., 21 Mar., 25 Apr., and 6 June Plants were watered as needed and

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47 continuously liquid fed with N at 150 mgL-1 using a 20.0N-4.4P-16.6K fertilizer (Peters 20-1020 Florida Special, The Scotts Co., Marysville, OH). Plants were observed daily and the date for first open flower and marketability were recorded. At marketabil ity, plant height was measured. Once all plants had reached first flower, flower height and width was m easured using calipers, and plants were evaluated for presence of powdery mildew (see exp. 1 for explanation of data collection). Temperature and light levels were recorded continuously over the length of the experiment using a HOBO Microstation Data Logge r (Onset Computer Cor p., Bourne, MA), and average daily minimum, maximu m, and 24-hr temperatures a nd daily light integral were determined for each season (Table 2-2). The experimental design was a split plot, with season as the main plot and cultivar as the sub-plot. Within each season, all cultivars were grown on a single bench in a complete random block design with three blocks. The late winter season had 9 plants and the other three seasons had 12 plants as the experimental unit. Data were analyzed using PROC MIXED in SAS 9.1 (SAS Institute, Cary, NC) and mean separation was performed using Tukeys at =0.05. Results and Discussion Experiment 2-1 Days to first flower Angelonia a ngustifolia cultivars flowered, on average, 52.5 days after pinch (DAP). The earliest-flowering cultivar was AngelMist Dark Lavender at 41.4 d and the latest was Angelface Blue Bicolor at 63.8 d (Table 2-3), a difference of almost 23 d. Cultivars within the Serena series were the mo st uniform and the range between the earliest and latest-flowering cultivar was approximately 1 d. The range between the first and last cultivar to flower in the Angelface and Angelina series were 10 d each. The AngelMist series had the widest range of flowering times, 18 d from the first to the last cultivar to flower. All Angelface

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48 and Serena cultivars took longer to flower than the average, and Angelina cultivars flowered more quickly than the average. The uniformity observed in the Serena series is consistent with the fact that it is seed propagated and the other three se ries are vegetatively propagate d. Plants of a vegetativelypropagated cultivar are very uniform, but cultiv ars within a vegetatively-propagated series usually have more genetic variab ility than seed propagated cultiv ars within a series (personal observation). These differences can have a great effect on crop production. Miller and Armitage (2002) reported that five cultivars within the AngelMist series responded similarly to temperature, irradiance, photoperi od, and growth retardants. Ho wever, Expt. 2-1 contained 15 cultivars from that series and they all did not flower uniformly. The wide variability seen between cultivars in the AngelMist series relative to the other two vegetatively-propagated series may be due to the fact that 15 cultivars were trialed in this experiment while the others had six cultivars each. Days to marketability Cultivars reached m arketability between 45.4 and 69.8 DAP, a range of more than 24 d (Table 2-3) and averaged 58.3 d. AngelMist Dark Lavender and Angelface Blue Bicolor were the earliest and latest to marketability, respectively. This is the same as observed for time to flower. The degree of uniformity within each series followed the same trends observed with time to first flower. All cultivars progressed from first flower to marketability in 4 to 8 d. Serena cultivars flowered the quickest after reaching first flow er, and Angelina Blue, Angelina Violet and White, AngelMist Dark Rose, and AngelMist Pi nk required the longes t length of time to reach marketability after starting to flower. This information w ill allow growers to monitor crop

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49 development and schedule shipment of the finished product for approximately 1 week after the start of flowering. Production guidelines for angelonia recomm end scheduling 7 to 8 weeks for Angelface cultivars (PW, 2006), 8 to 9 weeks for Angelin a cultivars (Selecta, 2008), 7 to 9 weeks for AngelMist cultivars (BFP, 2006), and 9 to 10 week s for Serena cultivar s (PanAm, 2006) grown in 2.2 L (1 gal.) pots. However, this length of time may vary from between 6 and 10 weeks depending upon time of year and greenhouse envir onment (Schoellhorn and Alvarez, 2002). In this experiment, crop production time varied fr om 6.5 to 10 weeks. AngelMist and Serena cultivars reached marketability within their anticipated cr op windows. Angelina cultivars finished one week earlier and Angelface cultivar s finished two weeks later than anticipated. These variations from the publis hed guidelines may be due to di fferences in the climate and environment where cultivars were trialed before commercial release. Ange lonia will finish more quickly when grown under warmer temperatures and higher light, up to an optimum, and the reverse when grown cooler and under lo wer light (Miller and Armitage, 2002). Plant height and width at marketability Plant height ranged from 14.0 cm for AngelM ist Basket Pink to 64.5 cm for Angelface Blue Bicolor (Table 2-4), a nd the average height was 39.0 cm The three shortest were AngelMist Basket cultivars, a sub-group of cultiva rs within the AngelMist series with trailing growth habits rather than the upr ight or mounding habits observed in most other cultivars (Fig. 21). Plant width ranged from 28.0 cm for A ngelMist Dark Lavender to 54.0 cm for Angelface Blue Bicolor and the average widt h was 41.5 cm. Angelface Blue Bicolor was both the tallest and widest cultiv ar. It would have had a much narrower width except for the fact that it began to lodge in the latter stages of production. It was also the latest cul tivar to flower

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50 and thus had the longest number of days to grow before data collection. The other very wide cultivars had mounding to semi-trailing growth ha bits, for example in AngelMist Basket Pink and Angelina Pink and White. The narrowest cu ltivars can be grouped into two categories: 1) cultivars that flower extremely early and thus did not have as much gr owing time before data collection, such as AngelMist Da rk Lavender, and 2) cultivars w ith very tall, upright growth habits, such as Angelface Pink. The growth habit of a plant is an importa nt consideration duri ng crop production. If a plant is extremely tall or wide, it will be difficult to grow it successfully. Plants with tall growth habits will not fit well on shipping racks and plan ts with wide, sprawling growth habits will be difficult to grow on tight spacing in a greenhouse. Cultivars with one or both of these characteristics will likely benefit from the application of a plant growth regulator during production. However, these are traits which a breeder may be able to select against when developing new cultivars. Inflorescence number at marketability At m arketability, the cultivars with the leas t and greatest number of inflorescences were Angelina Violet and White and AngelMist La vender with 2.3 and 4.2, respectively (Table 24). The average number of inflorescences acro ss all cultivars was 3.3. This measurement gives an idea of the appearance of the plant at marketability. Cultivars that have fewer inflorescences with open flowers at marketability have a c ouple of well-developed inflorescences. Cultivars with more inflorescences at marketability have fewer open flowers per inflorescence. Growers will require longer production times to produce cu ltivars like Angelface White and will require a longer period of time for it to develop a sufficient number of in florescences and a good display of color. However, the one advantage seen in Angelface White relative to AngelMist Dark

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51 Lavender is that it has larger flowers on its in florescences and will require fewer flowers to produce a similar display of color. Inflorescence number at 9.5 weeks The num ber of inflorescences per plant wa s determined at 9.5 weeks following pinch. This is different from the determination of the number of inflorescences at market because this will give an idea of its display of color after a sp ecified point in time. The range for number of inflorescences per plant at 9.5 weeks after pi nch was from 2.1 for Angelface Dresden Blue Bicolor to 29.2 for AngelMist Dark Lavender (Table 2-5), and the average was 12.6 inflorescences. Both time to flower and genotype are important components that help determine the number of inflorescences with open flowers at a specific point in time. Cultivars that naturally take longer to flower, such as Angelface Pink (refer to Table 2-3) will have fewer inflorescences in color than cultivars that natu rally flower earlier, su ch as AngelMist Dark Lavender given the same set of environmental c onditions (Fig. 2-2). Al so, the taller cultivars that have naturally upright growth habits, such as Angelface White and Angelface Purple Stripe, tend to have fewer inflorescences per pl ant than those with comp act or spreading growth habits, such as AngelMist Dark Lavender and AngelMist Lavender Stripe. Cultivars that have naturally tall, narrow growth habits and take longer to flow er, such as Angelface White, relative to compact, early-flowering cultivars, such as AngelMist Dark Lavender, have the undesirable characteristics for both traits. Flower size Flower size was m easured at 9.5 weeks following pinch. Flower height and flower width are positively related to one anothe r, and taller flowers tend to ha ve larger widths than shorter flowers. Flowers, in general, ar e round or oval in shap e (Fig. 2-3). In cert ain flowers, the side petals reflex backward more than others. This backward bending of the flower causes a decrease

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52 in overall flower width and decreases the amount of visual impact created by the flower. Larger flowers tend to have a rounded shape and smaller fl owers tend to have an oval shape. Very little petal reflex is observed in the Angelface series and the AngelMist Basket cultivars. Medium sized flowers, such as those on AngelMist Purple Stripe, tend to also be round in shape, but will have a slight more degree of petal reflex th an seen in the largest-flowering cultivars. Cultivars with small flowers, such as the Serena series or AngelMist Dark Lavender, tend to exhibit the greatest degree of petal reflexing. Flower height ranged from 20.1 mm in AngelMist White Improved to 30.7 mm in Angelface Dresden Blue. Flower width ranged from 12.7 mm in Angel Mist Pink to 28.4 mm in Angelface Blue. Average flower height and width were 23.8 a nd 20.0 mm, respectively. Cultivars in the Angelface series had the tallest a nd widest flowers. Cult ivars in the other three series did not group together very well, but all of the Serena cultivars had flowers shorter and narrower than the overall averages (Fig. 2-4). In general, cultivars with tall growth habits tend to have larger flowers than the cultivars with compact growth habits. Exceptions include the AngelMist Basket cu ltivars, with trailing growth habits, and AngelMist Purple Improve d, which has a medium growth habit but relatively large flowers. Powdery mildew index Powdery m ildew susceptibility was not one of the initial traits of interest in this experiment. However, during the cultivar scr een, it was a natural occurrence in the greenhouse, and was added as a trait of interest since cultivar s exhibited varying levels of sensitivity. Plants were not inoculated for this study, and no fungicides were applied for the duration of the experiment so that the degree of susceptibility could be quantified. Plants were evaluated on a 15 rating at the end of the experiment, and suscep tibility ranged from hi ghly resistant to highly

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53 susceptible. The most susceptible cultivar, w ith an average rating of 5.0, was AngelMist White Improved (Table 2-5). The least susceptible cu ltivars, with ratings of 1.0, were Angelface Blue, Angelface Blue Bicolor, Angelina Violet and White, AngelMist Basket Pink, AngelMist Pink, AngelMist Dark Rose, and Serena Lavender Pink. Over half of the cultivars in this stud y had ratings less than 2.0, which would be considered saleable. Anythi ng with greater than a 2.0 rati ng would have benefited from a fungicide spray in order to reta rd the spread of the pathogen from the lower leaves up through the plant canopy. Cultivars with a high degree of susceptibility should be closely monitored during production to prevent an outbreak from affecting plant quality. The range of susceptibility observed indicates that powdery mi ldew tolerance might be a trait that breeders could select for in cu ltivar development. Experiment 2-2 Days to first flower The season x cultivar interaction was significan t for number of days to first open flower and number of days to marketability (Table 2-6). The mean separations for each parameter were very similar, so the discussion will focus on number of days to first flower. Generally, the number of days to first open flower decreased as the seasons progressed from late winter (season 1) to early summer (seas on 4) (Table 2-7). The difference in days to flower between season 1 and season 4 was the leas t (17 d) in AngelMist Dark Lavender and the greatest (almost 29 d) in Angelface Blue. This is a difference of 2 to 4 weeks in production from late winter to early summer. AngelMist Dark Lavender was the quickest to flower in all four seasons. Angelface White and AngelMist Purple Improved were th e slowest to flower in seasons 1 and 2, and Angelface White was the slowest in seasons 3 and 4. The range between the first and last

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54 cultivar to flower was smallest in season 4, at 16.2 days. A 2-week difference in production time can greatly impact the number of turns in a greenhouse during the spring production season if a late-flowering cultivar is grow n instead of an early-flowerin g cultivar. This additional production time may be justified, however, if it is a unique cultivar and can be sold at a higher price. Temperature and light, both photoperiod a nd irradiance, has been shown to play important roles in flower initia tion and development. Angelonia is a day-neutral species (Miller and Armitage, 2002; Starman, 2001) so irradiance and total light are more important variables than photoperiod for this crop. Irradiance play s a role in floral in itiation, and increased irradiance, up to an optimum, w ill hasten floral initiation (Kaczp erski et al., 1991). As daylength increases, the DLI, or total irradiance captured in a 24-h period, will increase due to a longer day length from which to capture light energy (Korczynski et al., 2002) Temperature is involved in the rate of development of the floral buds. Since floral development is a metabolicallycontrolled process, an increase in temperature will hasten development up to an optimum, and then retard development at supraoptimal te mperatures (Kaczperski et al., 1991). Increased irradiance hastened flowering by 5 d in Achillea and 7 days in Gaura when grown at a constant 22 C (Fausey et al., 2005). Increased averag e daily temperature hastened flowering in Campanula carpatica Blue Clips (Nui et al., 2001), and pansy Universal Violet exhibited a linear increase in the rate of pr ogress to flowering as temperatur e increased up to an optimum of 21.7 C, resulting in earlier flower ing at higher temperatures (Adams et al., 1997). Increases in both temperature and irradiance hastened flowering in two Gypsophila paniculata cultivars (Hinkleton et al., 1993) and in Pelargonium xhortorum Radio (Welander, 1983).

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55 These results for angelonia are consistent with results from Miller and Armitage (2002), in which the number of days to visible bud and days to flower decreased with an increase in either irradiance or temperature. In their work each va riable was examined in separate experiments so it is unknown if there is an interaction between te mperature and irradiance in angelonia. For the work reported here, the average daily temperatur e increased from 21.4 C in season 1 to 28.0 C in season 4. The daily light integr al (DLI) increase d from 20.2 molm-2d-1 in season 1 to 21.5 molm-2d-1 in season 2, then declined to 20.6 and 17.9 molm-2d-1 in seasons 3 and 4, respectively. Although irradiance de clined in seasons 3 and 4, the number of days to flower still decreased. This may indicate that temperature has more influen ce on flowering than light above a minimum light level. The light levels observed in this experiment (17.9 to 21.5 molm-2d-1) were relatively high compared to those observed in greenhouses in northern states in early spring (typically less than 10 molm-2d-1) (Korczynski et al., 2002) and ma y not have been the critical factor influencing flower time. Plant height The cultivar x season interaction was signifi cant, and the general trend observed was an incre ase in plant height at marketability from season 1 to season 4 in all cultivars (Table 2-8). AngelMist Purple Improved had the least difference in height between seasons (7 cm) and was the only cultivar in which plan t height was not significantly di fferent across season. AngelMist Purple Stripe had the greatest increase in plant height between seasons (26 cm). Within the different seasons, AngelMist Dark Lavender was the shortest cultivar in every season. AngelMist Purple Stripe, Angelface White, and Angelface Blue were the tallest cultivars in season 1, and AngelMist Purple Stripe and Angelface White were the tallest in seasons 2 through 4. The range between the tallest and shortest cultivars was 33 to 35 cm in seasons 1 through 3, but increased to 45 cm in season 4.

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56 In the study conducted by Miller and Arm itage (2002), looking at the effects of increasing temperature or irradian ce separately, it was reported that an increase in temperature had a greater influence on plant he ight than an increase in irradian ce. In their experiments, plant height increased by almost 17 cm when grown at a constant 30 C compared to 22 C, and plants grown at 30 C had longer internodes and more br ittle stems than those grown at 22 C. Plant height in the highest irradiance le vel was only 3 cm shorter than thos e grown at the lowest level. An increase in both temperature and irradiance sh ould have opposing influences on plant height, but temperature appears to have a greater influence on final plant height than irradiance, especially under higher light levels. Increased plant growth in the la ter seasons will require growers to use additional height control measures. Flower size The analysis for this param eter was based on only six cultivars due to missing data for Angelface White in season 4. At the time of da ta collection, not enough plants had mature, open flowers for accurate measurement of the flower height. Since the results for flower height and flower width were very simila r, only the results for flower he ight will be presented. The interaction was not significant, but the main effect of season and the main effect of cultivar were significant. The lack of an intera ction indicates that flow er size in all cultivars responded similarly to environmental changes with season while other growth parameters showed differences in cultivar responses to se ason. The differences in flower size for the cultivars are consistent with the results observe d in Expt. 2-1, and is not surprising considering that the flower size was one parameter us ed to select the cultivars for this study. Flower size has been shown to decrease as temperature increases in chrysanthemum (Willits and Bailey, 2000); calendula, impatiens, mimulus, and torenia (Warner and Erwin, 2005a); and pansy (Warner and Erwin, 2006). However, in this study on angelonia, the range

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57 between seasons was only 1.5 mm, a very small amount Flower height was greatest in season 1, but no significant differences were observed between seasons 2 through 4 (Table 2-9). Angelface Blue had the tallest flowers (average of 29.4 mm for the 4 seasons), followed by AngelMist Purple Improved. AngelMist Dark Lavender had the shortest flowers (21.1 mm) and was significantly different from all cu ltivars except AngelMist Lavender Stripe. Powdery mildew susceptibility The season by cultivar interaction w as signi ficant for powdery mild ew susceptibility (Table 2-6). The level of susceptibility obser ved in Expt. 2-2 is similar to the level of susceptibility observed for the same cultivars in Expt. 2-1. The level of susceptibility for five of the cultivars was similar across season, and was low relative to the other cultivars (Table 2-10). AngelMist Lavender Stripe had a greater incidence of powdery mildew in seasons 2 and 4 relative to plants of the same cultivar grown in seasons 1 and 3. AngelMist Dark Lavender had a greater incidence of powdery mildew in seasons 1 through 3 relative to plants of the same cultivar grown in season 4. Under these e xperimental conditions, A ngelMist Dark Lavender was the most susceptible cultivar. In Florida, powdery mildew tends to be a more common occurrence in the spring months and less of an issue in the summer months. This is due to the fact that warm and dry environments are most favorable for spore germination and hyphal gr owth (Agrios, 2005). Conclusions In Expt. 2-1, Angelonia a ngustifolia cultivars exhibited a wide range of variability in all of the growth and flowering traits quantified. The number of days to first flower spanned a period of more than 3 weeks, a long time for a crop w ith a total production tim e of 6.5 to 10 weeks. Cultivars varied in plant height and width at marketability by more than 50 and 26 cm, respectively. Plants with shor ter, wider habits tended to ha ve more inflorescences open at

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58 marketability. At 9.5 weeks after pinch, once all the cultivars had started flowering, those with wider, spreading habits had more inflorescences than those with na rrower, upright growth habits. Cultivars with narrow, upright growth habits tend ed to have fewer inflorescences per plant but larger flowers. Powdery mildew susceptibility was highly variable amongst cultivars and ranged from highly resistant to highly susceptible. In Expt. 2-2, number of da ys to first open flower, plan t height, and powdery mildew susceptibility were all subject to season x cultivar inte ractions. Flower size was influenced by the main effects of cultivar and temperature. In the warmest season (season 4, early summer), plants flowered quickly, were tall, had the least susceptibil ity to powdery mildew, and had slightly smaller flowers than plants grown in la te winter. Cultivars varied widely in their response to changing temperature and irradiance levels. AngelMist Dark Lavender did not flower any quicker between seasons 2 and 4, but other cultivars, like Angelface Blue exhibited a continual decrease in the number of days to flower with each successive season. These results indicate that there is variability present betw een cultivars that may be useful in new cultivar selection. The results from Expt. 2-2 indicate that season ality is an important fa ctor to consider in commercial production. In southern states, early to late spring w ould be ideal for production. In late winter, this crop would need extra time to reach maturity and in early summer, plant quality would likely decrease. In northern states, the use of supplemental hea ting and/or lighting will help hasten crop maturity for the spring season.

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59 A B C Figure 2-1. Range of plant he ight present in commercial Angelonia angustifolia cultivars. A) AngelMist Purple Stripe B) Angelina Pi nk C) AngelMist Bask et Pink. Height data (Table 2-4) collected at marketab ility was 55.5, 38.5 and 14.0 cm, respectively. Pictures were taken at 9.5 weeks afte r pinch, and plant heights were 66.0, 39.0, and 20.5 cm, respectively. A B C Figure 2-2. Examples of the range on inflorescence number present in Angelonia angustifolia cultivars 9.5 weeks after pi nching. Plants were grown from January to April 2007. The cultivars and average number of inflor escences are as foll ows: A) AngelMist Dark Lavender with 29.2 inflorescences B) Angelina Pink and White with 15.7 C) Angelface Pink with 2.3.

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60 Figure 2-3. Range of flower size present in commercial Angelonia angustifolia cultivars. From left to right, the cultivars are Angelface Blue, AngelMist Purple Stripe, and Serena White. In many of the small-flow ered cultivars, the side petals reflex backwards to a greater degree than in larger flowers. Figure 2-4. Comparison between inflorescences in Angelonia angustifolia Angelface White (left) and Serena White (right). Note differences in inflorescence length, flower size, and the spacing of the fl owers along the inflorescence.

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61Table 2-1. Plant growth and flow ering characteristics for seven Angelonia angustifolia cultivars used in Expt. 2-2. Angelina Violet and White AngelMist Dark Lavender AngelMist Lavender Stripe AngelMist Purple Improved AngelMist Purple Stripe Angelface Blue Angelface White Growth habit Plant height Medium Short Medium Medium Tall Tall Tall Plant width Wide Medium Wide Medium Narrow Narrow Narrow Internode length Medium Shor t Medium Medium Long Long Long Branching Medium High High Medium Medium Medium Low Stem thickness Medium Thin Medium Medium Thick Medium Thick Leaf thickness Thin Thin Thin Medium Thick Medium Thick Leaf pubescence Glabrous Glabrous Glabrous Glabrous Pubescent Glabrous Pubescent Leaf stickiness Medium Low Medium Medium Very high Medium Very high Flowering Flower size Medium Small Medium Large Large Large Large Time to flower Medium Early Me dium Late Medium Medium Late Internode length Medium Shor t Medium Medium Long Long Long Inflorescence number per plant Medium High Medium Low Low Medium Low Flower number per inflorescence Medium High Medium Medium High High High Petal rolling of side petals High Hi gh Medium Medium Low Low Medium Flower color Bicolored purple upper petals and white lower petals Lavender Bicolored white petals with light lavender stripe down center Purple Bicolored white petals with purple stripe down center Blue with purple specks White

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62 Table 2-2. Temperature and light data for Expt. 2-2. Season 1 Season 2 Season 3 Season 4 Late winter (30 Jan 20 Apr) Early spring (21 Mar 18 May) Late spring (25 Apr 20 Jun) Early summer (6 Jun 23 Jul) Daily max T (C) 28.0 31.7 34.4 36.2 Daily min T (C) 17.0 17.7 19.6 22.7 24-hr average (C) 21.4 23.5 25.7 28.0 DLI (molm-2d-1)z 20.2 21.5 20.6 17.9 z DLI = daily light integral

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63 Table 2-3. Angelonia angustifolia flowering data for 31 cultivars (Expt. 2-1). Marketability was defined as 20 open flowers per plant. Anal ysis of variance for both parameters were significant at P<0.001. Cultivar First open flower (days from pinch) Marketability (days from pinch) Angelface Blue 54.0 58.8 Angelface Blue Bicolor 63.8 69.8 Angelface Dresden Blue 60.8 66.0 Angelface Pink 59.8 66.4 Angelface Wedgewood Blue 56.1 59.9 Angelface White 59.4 64.9 Angelina Blue 49.3 56.5 Angelina Dark Blue 42.5 49.4 Angelina Pink 47.4 53.5 Angelina Pink and White 52.4 57.3 Angelina Violet and White 49.1 57.0 Angelina White 47.9 53.9 AngelMist Basket Pink 54.1 58.6 AngelMist Basket Purple 52.4 57.7 AngelMist Basket White 47.4 53.4 AngelMist Dark Lavender 41.4 45.4 AngelMist Dark Pink 52.6 57.0 AngelMist Dark Rose 48.3 55.8 AngelMist Deep Plum Imp. 53.9 57.4 AngelMist Lavender 47.4 53.0 AngelMist Lavender Stripe 44.6 53.1 AngelMist Pink 49.6 57.1 AngelMist Plum 57.4 62.8 AngelMist Purple Imp. 59.6 64.6 AngelMist Purple Stripe 54.3 58.8 AngelMist White Cloud 44.1 50.4 AngelMist White Imp. 50.7 57.0 Serena Lavender 57.2 63.9 Serena Lavender Pink 56.1 61.4 Serena Purple 57.2 63.8 Serena White 56.4 62.8 Analysis of variance *** *** Waller-Duncan MSD =0.05 2.5 2.4

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64 Table 2-4. Plant data for 31 cultivars of Angelonia angustifolia collected at marketability (Expt. 2-1). Marketability was defined as the first date at which a plant had 20 or more open flowers. Analysis of variance for a ll parameters were significant at P<0.001. Cultivar Plant height (cm) Plant width (cm) Inflorescence number Angelface Blue 52.0 41.5 3.2 Angelface Blue Bicolor 64.5 54.0 3.2 Angelface Dresden Blue 56.0 36.5 3.0 Angelface Pink 48.5 36.5 2.7 Angelface Wedgewood Blue 49.5 36.5 3.0 Angelface White 59.5 39.5 2.6 Angelina Blue 40.0 44.0 3.3 Angelina Dark Blue 31.0 42.0 3.1 Angelina Pink 38.5 39.0 3.5 Angelina Pink and White 42.5 44.5 3.6 Angelina Violet and White 44.0 43.0 2.3 Angelina White 32.0 35.5 3.6 AngelMist Basket Pink 14.0 53.5 3.2 AngelMist Basket Purple 20.0 53.0 3.2 AngelMist Basket White 23.5 47.0 2.7 AngelMist Dark Lavender 26.0 28.0 3.6 AngelMist Dark Pink 39.5 46.5 3.6 AngelMist Dark Rose 35.0 36.5 4.0 AngelMist Deep Plum Imp. 45.5 36.5 2.4 AngelMist Lavender 31.0 36.0 4.2 AngelMist Lavender Stripe 36.0 42.5 3.6 AngelMist Pink 33.5 40.0 3.8 AngelMist Plum 45.0 37.0 3.4 AngelMist Purple Imp. 44.0 38.5 3.2 AngelMist Purple Stripe 55.5 43.5 3.2 AngelMist White Cloud 32.0 36.0 2.9 AngelMist White Imp. 39.0 44.0 3.4 Serena Lavender 34.5 41.0 3.8 Serena Lavender Pink 33.0 41.0 3.9 Serena Purple 32.5 39.5 4.0 Serena White 36.0 47.0 3.9 Analysis of variance *** *** *** Waller-Duncan MSD =0.05 3.5 4.5 0.8

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65 Table 2-5. Angelonia angustifolia plant data for 31 cultivars co llected 9.5 weeks after pinch (Expt. 2-1). Powdery mildew was rated fr om 1 to 5, with 1 = least incidence of powdery mildew and 5 = most severe incidence of powdery mild ew. Analysis of variance for all parameters was significant at P<0.001. Cultivar Number of inflorescences Flower height (mm) Flower width (mm) Powdery mildew indexz Angelface Blue 4.2 30.3 28.4 1.0 Angelface Blue Bicolor 2.1 24.6 23.2 1.0 Angelface Dresden Blue 2.9 30.7 27.3 1.1 Angelface Pink 2.3 26.4 24.5 1.1 Angelface Wedgewood Blue 4.1 28.7 25.9 3.6 Angelface White 2.3 29.4 26.3 1.1 Angelina Blue 17.3 22.0 18.8 4.5 Angelina Dark Blue 16.1 24.5 21.0 2.8 Angelina Pink 19.6 21.6 18.7 3.3 Angelina Pink and White 15.7 21.7 19.6 4.3 Angelina Violet and White 20.3 22.6 19.6 1.0 Angelina White 18.6 24.1 16.6 2.3 AngelMist Basket Pink 8.7 23.6 21.0 2.9 AngelMist Basket Purple 10.9 26.3 24.3 1.8 AngelMist Basket White 12.2 24.3 21.3 1.0 AngelMist Dark Lavender 29.2 20.7 18.4 4.7 AngelMist Dark Pink 11.6 25.3 19.3 1.1 AngelMist Dark Rose 19.2 22.6 14.5 1.0 AngelMist Deep Plum Imp. 5.6 23.7 22.1 1.6 AngelMist Lavender 27.6 21.0 15.3 2.2 AngelMist Lavender Stripe 24.7 22.5 19.3 2.6 AngelMist Pink 21.1 21.9 12.7 1.0 AngelMist Plum 4.1 22.5 13.2 4.4 AngelMist Purple Imp. 2.9 25.0 22.6 1.3 AngelMist Purple Stripe 4.8 24.7 22.1 1.1 AngelMist White Cloud 26.9 21.1 14.6 2.7 AngelMist White Imp. 24.3 20.1 14.4 5.0 Serena Lavender 5.3 21.7 19.6 2.0 Serena Lavender Pink 8.2 22.6 19.1 1.0 Serena Purple 7.2 22.0 18.0 1.3 Serena White 9.7 20.6 17.1 2.0 Analysis of variancey *** *** *** *** Waller-Duncan MSD =0.05 3.4 1.6 1.4 0.8

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66Table 2-6. Split plot analysis of variance for seven Angelonia angustifolia cultivars grown at four different seasons from January to July 2007 (Expt. 2-2). Plant height was collected as each plan t reached marketability. Powdery mildew index and flower height data were collected on the same da y once all cultivars reached marketability. Days to first flowery Days to marketability Plant height (cm) Powdery mildew index Flower height (mm) Season *** *** *** *** ** Cultivar *** *** *** *** *** Seas. x Cv. *** *** *** *** NS y NS, *, **, and *** non-significant or significant at P 0.05, 0.01, and 0.001, respectively. Table 2-7. Number of days to first flower in seven Angelonia angustifolia cultivars across four seasons (Expt. 2-2). The dates for each season were the following: late winter 30 Jan to 20 Apr; early spring 21 Mar to 18 May; late spring 25 Apr to 20 Jun; and early summer 6 Jun to 23 July 2007. Uppercase letters indica te mean separation within each column (between seasons within each cultivar) and lowercase letters indicate mean separation within each ro w (between cultivars within each season). Cultivar Season Angelina Violet and White AngelMist Dark Lavender AngelMist Purple Imp. AngelMist Lavender Stripe AngelMist Purple Stripe Angelface White Angelface Blue 1 Late winter 49.1 A c 41.4 A d 59.6 A a 44.6 A d 54.3 A b 59.4 A a 54.0 A b 2 Early spring 41.3 B bc 26.3 B e 46.8 B a 38.3 B cd 41.9 B b 49.5 B a 36.4 B d 3 Late spring 35.7 C c 27.6 B e 39.5 C b 31.8 C d 39.5 B b 46.9 B a 29.5 C de 4 Early summer 28.8 D bc 24.2 B d 32.3 D b 26.1 D cd 32.1 C b 40.4 C a 25.2 D d

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67Table 2-8. Plant height (cm) at marketability in seven Angelonia angustifolia cultivars across four seasons (Expt. 2-2). Marketability was defined as the first date at which a plant had 20 or more open flowers. The dates for each season were the following: late winter 30 Jan to 20 Apr; early spring 21 Mar to 18 May; late spring 25 Apr to 20 Jun; and early summer 6 Jun to 23 July 2007. Uppercase letters indicat e mean separation within each column (b etween seasons within each cultivar) and lowercase letters indicate mean separation within each row (between cultivars within each season). Cultivar Season Angelina Violet and White AngelMist Dark Lavender AngelMist Purple Imp. AngelMist Lavender Stripe AngelMist Purple Stripe Angelface White Angelface Blue 1 Late winter 44.0 B b 26.0 B d 44.0 A b 36.0 B c 55.5 C a 59.0 C a 52.0 AB a 2 Early spring 50.0 AB b 32.5 AB c 48.5 A b 46.5 A b 63.0 BC a 67.0 B a 51.5 B b 3 Late spring 52.0 A bc 38.0 A d 47.0 A c 45.5 A c 69.0 B a 73.5 AB a 58.0 AB b 4 Early summer 53.0 A bc 37.0 A d 51.0 A c 51.0 A c 81.5 A a 78.0 A a 59.0 A b Table 2-9. Flower height (mm) in six Angelonia angustifolia cultivars across four seasons (Expt. 2-2). Data was collected once all cultivars reached marketability. Angelface White had missi ng data for the early summer season and was non-estimable, creating an unbalanced factorial. Data were analyzed for the remaining six cul tivars with complete data sets for all seasons. The dates for each season were the following: late winter 30 Jan to 20 Apr; early spring 21 Mar to 18 May; late spring 25 Apr to 20 Jun; and early summer 6 Jun to 23 July 2007. Cultivar Season Angelina Violet and White AngelMist Dark Lavender AngelMist Purple Imp. AngelMist Lavender Stripe AngelMist Purple Stripe Angelface Blue Pooled mean across cultivar 1 Late winter 22.6 23.0 25.0 22.5 24.7 30.3 24.7 a 2 Early spring 22.1 21.0 24.3 21.1 23.4 29.7 23.6 b 3 Late spring 21.3 20.0 24.2 20.8 22.6 28.6 22.9 b 4 Early summer 21.3 20.4 23.6 20.3 22.3 29.1 22.9 b Pooled mean by cultivar 21.8 d 21.1 e 24.3 b 21.2 de 23.3 c 29.4 a

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68Table 2-10. Powdery mildew index in seven Angelonia angustifolia cultivars across four seasons (E xpt. 2-2). The index ranged from 1 to 5, with 1 indicating the least incide nce of powdery mildew and 5 indicating a severe incidence of powdery mildew. The dates for each season were the following: late winter 30 Jan to 20 Apr; early spring 21 Mar to 18 May; late spring 25 Apr to 20 Jun; and early summer 6 Jun to 23 July 200 7. Uppercase letters indicate mean separation within each column (between seasons within each cu ltivar) and lowercase letters indicate me an separation within each row (between cultivars within each season). Cultivar Season Angelina Violet and Whitex AngelMist Dark Lavender AngelMist Purple Imp. AngelMist Lavender Stripe AngelMist Purple Stripe Angelface White Angelface Blue 1 Late winter 1.0 A c 4.7 A a 1.3 A c 2.6 A b 1.1 A c 1.1 A c 1.0 A c 2 Early spring 1.1 A b 5.0 A a 1.4 A b 1.7 B b 1.3 A b 1.0 A b 1.0 A b 3 Late spring 1.2 A c 4.6 A a 1.2 A c 2.2 A b 1.2 A c 1.0 A c 1.5 A c 4 Early summer 1.0 A b 1.9 B a 1.0 A b 1.0 B b 1.0 A b 1.0 A b 1.0 A b

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69 CHAPTER 3 PLANT DATE X CULTIVAR INTERACTIONS INFLUENCE SUMMER LANDSCAPE PERFORM ANCE OF Angelonia angustifolia Introduction High tem perature stress in plants can lead to a reduction in flower number, an increase in floral bud abortion, or complete cessation of flowering. The mechanism underlying this temperature response may be related to either acute or chronic high temperature stress depending upon species. In greenhouse production, decreased flower bud number has been observed in pansy and petunia (Kaczperski et al., 1991; Warner and Erwi n, 2006) In field crops, high temperature stress has been shown to reduce flower number in Capsicum annuum (Erickson and Markhart, 2001) and reduce yield in tomato and ca nola (Morrison and Stewart; Sato et al., 2000). Warner and Erwin (2005b) published a model for floral abortion in Arabidopsis thaliana genotypes and indicated that plant response was related to a thre shold number of hours above a critical temperature. For landscape beds and other large-scale planting s, it is critical to select plants that will flower throughout the season. Angelonia angustifolia is a plant with reported heat tolerance but has been observed to go out of flower duri ng the summer months in Florida (personal observation). Different cultivars will stop flowering at different times during the summer and remain without flowers for varying lengths of time. The objective of this research was to determine the effect of plant date and cultivar on landscape performance and flowering of eight angelonia cultivars. Materials and Methods See Experim ent 2-2 for a descri ption of materials and methods related to th e production of rooted cuttings. This experiment consisted of the seven cultivars shown in Table 2-1 and Serena

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70 White planted at three different times into a land scape bed. Rooted cuttings of all cultivars were planted into 80 mm Ellepots (Knox Nursery, Winter Park, Florida) on 14 Mar., 18 Apr., and 16 May 2007, respectively, and grown in a natural vent ilated greenhouse for 3 weeks. They were pinched two weeks after planting, and twelve plants of each cultivar were transplanted into an inground landscape bed (Gainesville, FL) on three pl ant dates spaced at 4week intervals, on 4 Apr., 2 May, and 30 May 2007, respectively. The bed was amended with 122 gm-2 Osmocote 18-6-12 (18.0N-2.6P-10.0K) slow-release fertilizer with 6 to 9 month release rate (The Scotts Co., Marysville, OH), and plants were irrigated as needed with drip irrigation. This experiment was designed as a split plot, with plant date as the main plot and cultivar as the subplot. Within each plant date, cultivars were planted in a complete random block design consisting of three blocks and four plants per block. Plants were planted 46 cm apart within each row and 61 cm between rows. Data were collected at 10 weeks after transpla nt (WAT) on two plants of each cultivar per block (13 Jun., 11 July, and 8 Aug., respectively) and included plant height, inflorescence number, and flower height. Plant height was me asured from the soil surface to the tallest point of the plant. Inflorescence number was determ ined by counting all inflorescences containing at least one open flower. Flower height was measured with calipers. Beginning 5 WAT, plants were evaluated every two weeks and a flower ra ting of 0-10 was determined for each row since all plants within each cultivar row flowered uni formly. A flower rating of 3 was determined to be the minimum critical value for acceptable landscape performance. The final evaluation date for all plants was 10 Oct. Temperature and DL I was recorded every 15 min. by a weather station (Hortimax USA, Inc., Rancho Santa Margarita, CA) and daily average minimum, maximum, and 24-h temperatures and DLI were calculated. Monthly averages can be found in Table 3-1.

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71 Data were analyzed using SAS 9.1 (SAS Institute, Cary, NC) using the PROC MIXED procedure. The flower ratings were analyzed two different ways: 1) a comparison based upon plant age, and 2) a comparison based upon the date when evaluations were collected. Evaluations based upon plant age were used to dete rmine if plants stopped flowering at a similar physiological age, and evaluations based upon date collected helped determine the amount of flowering present in cultivars at the same point in time. Results and Discussion 10-Week Data Plant height The plant da te x cultivar inte raction was significant for plan t height as measured 10 weeks after transplant (WAT) (Table 3-2). Plants were evaluated on 13 Jun., 11 July, and 8 Aug. 2007, respectively. This evaluation interval was c hosen because it provided adequate time for all cultivars to begin flowering (see Chapter 2) Angelina Violet and White and AngelMist Purple Improved were the only two cultivars to have significant hei ght differences between plant dates (Table 3-3). In these two cultivars, plant height increas ed from plant date 1 (PD 1) to PD 2, then decreased from PD 2 to PD 3, but only PD 2 and PD 3 were significantly different from each other. The difference in height between plant dates can be attributed to a combination of factors, including temperature and the numbe r of inflorescences per plant. In PD 1, average temperatures were cooler than in PD 2 and PD 3, so the plants developed slower. Als o, angelonia plants have been shown to develop longer internodes under higher temperat ures (Miller and Armitage, 2002). In PD 3, these two cultivars, Angelin a Violet and White and AngelMist Purple Improved, had very few inflorescences per plant, and the inflorescence can represent a significant portion of the total plant height due to its raceme architecture (Chapter 1). When a

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72 cultivar has no inflorescences, or very few developed ones, the plant will have a shorter overall plant height than plants with well-developed inflorescences. In PD1, AngelMist Purple Stripe and A ngelface White were taller than all other cultivars except for Angelface Blue. In PD2 and PD3, AngelMist Purple Stripe and Angelface White were the tallest cultivars. Cultiv ars that are tall in production will tend to be tall landscape plants and compact plants in produ ction will be compact plants in the landscape. Inflorescence number The plant da te x cultivar inte raction was significant for inflorescence number (Table 3-4). Angelina Violet and White, AngelMist Dark Lavender, and AngelMist Lavender Stripe had significantly less flowers in PD 2 and 3 relative to PD 1. Serena White and AngelMist Purple Improved had significantly less flowers in PD 3 compared to PD 1 and 2. AngelMist Purple Stripe, Angelface White, and Angelface Blue showed no significant difference in inflorescence number across plant dates. Serena White had the highest number of inflorescences in PD 1 and 2, and Angelface Blue and AngelMist Purple Improved had the least number of inflorescences, respectively, in PD 1 and 2. In PD 3, there were no significant differences between cultivars. These results indicate that all cultivar s except AngelMist Purp le Stripe, Angelface White, and Angelface Blue had a lower number of inflorescences at the later plant dates and were more sensitive to the increase in temperatur e. In addition to having fewer inflorescences, the plants also had fewer open flowers per in florescence in the later plantings (data not presented). It has been reported that plants su bjected to high temperature stress will have fewer flowers than those not subject ed to it (Morrison and Stew art, 2002; Sato, 2000). Although Angelface Blue did not have a significant differe nce between plant dates, it never had a large

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73 display of color relative to the other two cultivars that were unaffected by plant date. Angelface Blue had 11, 5, and 1 inflorescences, respectively, across plant dates. The greatest amount of variabil ity between cultivars occurred in PD 1, and the least amount of variability occurred in PD 3. The range between cultivars with the greatest and least number of inflorescences was 160, 127, and 29 in PD 1 through PD 3 In PD 1, growing temperatures were ideal for cultivars to flower at their phenotypic potenti al, and peak flowering occurred approximately 10 WAT (see discussion below). In successive plantings, the average daily temperatures increased while the DLI remained relatively consistent. However, in the later plantings, the increase in temperature resulted in plants in which flowering was inhibited due to higher than optimal temperatures for continual inflorescence development A study by Miller et al. (2001) indicated that the optimal temperat ure for net photosynthe sis in two angelonia cultivars was approximately 20 C. Above that temperature, gross photosynthesis continued to increase but was negated by an increase in da rk respiration. Under our conditions during the summer, it can be expected that flowering will be affected by the high average temperatures observed relative to the optimal temperat ure for photosynthesis in this species. Flower height The plant da te x cultivar inte raction was significant for flow er height (Table 3-5). The only cultivar with a significant change in fl ower size across plantings was AngelMist Purple Improved. The mean flower height for this cultivar in each PD were 21.5, 16.6, and 17.9 mm, respectively, with PD2 significantly smaller than PD 1. In PD 1, Angelface Blue had taller flowers than all cultivars except Angelface White. In PD 2, Angelface Blue, Angelface White, a nd AngelMist Purple Stripe had the tallest flowers, and in PD 3, Angelface White had the tallest flowers. The range between the largest and smallest flowers was 8 to 9.2 mm between plant dates.

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74 In many species a decrease in flower size has been observed with an increase in temperature, including chrysanthemum, calendu la, impatiens, mimulus, torenia, and pansy (Warner and Erwin, 2005a, 2006; Willits and Baile y, 2000). In angelonia, it appears that the primary response to high temperature is a decr ease in the number of in florescences and not a decrease in flower size, whereas in pansy, th e tradeoff under high temperatures is a similar number of flowers, but a decrease in flower size (Warner and Erwin, 2006). Flower Ratings by Plant Age Plants were evaluated ev ery two weeks, starting at 5 WAT, for degree of flower coverage using a 10 point scale, with 0 indicating no in florescences and 10 indica ting 100 percent flower coverage. A rating of 3 was considered to be the minimum value for acceptable landscape flowering. Ratings were compared 1) on the basis of plant age and 2) by evaluation date. A comparison based upon plant age (similar WAT) allo ws for a comparison of cultivars at the same physiological age, but exposed to different environmental conditions. At each WAT comparison, successive plantings were gr own under higher average temperatures. Plants ratings were analyzed from 5 to 19 WAT and the plant date by cultivar interaction was significant for all intervals except for 19 WA T (Table 3-6). A grap hical representation of each cultivar for the 3 plant dates is given in Figu re 3-1. Cultivars varied in their response to the environmental conditions. For example, AngelM ist Dark Lavender, a h eat-sensitive cultivar had ratings of 7.7, 2.7, and 1.3 at 9 WAT for PD 1 to 3. In contrast, AngelM ist Purple Stripe, a heat-tolerant cultivar had ratings of 6.0, 5.0, and 4.7, respectively. Other cultivars, such as AngelMist Purple Improved had poor landsca pe performance regardless of plant date. In all cultivars, the highest flower ratings achieved by each cultivar occurred within PD 1. In PD 1, cultivars tended to maintain peak flow ering for a longer period of time than those from

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75 PD 2 or 3. An exception to this trend was AngelMist Purple Stripe in which all plant dates maintained flower ratings >3 for all evaluations after 5 WAT. In PD 1, most cultivars reached peak flowering around 11 WAT and then ratings generally declined after that. Plants in PD 3, however, never reached a peak flowering window, and maintained a relatively low amount of flow ering (less than or equal to 4) all summer. Regardless of the actual ratings, all plant da tes reach a maximum fl owering around 3 months after transplanting into the gardens, which coinci des with their designation as an annual crop. However, the temperature during initial establis hment will have a major impact on the flowering capability of a cultivar, with higher temper atures inhibiting inflorescence development. Flower Ratings by Evaluation Date Flower ratin gs were also compared by evaluation date. The first evaluation date for all three plantings was 4 July, once PD 3 had been in-ground for 5 weeks, and ended on 10 Oct. This provided a comparison of how the different plant dates responded in the landscape at specific points in time, regardless of plant age. The plant date by cultivar interaction was significant on 4 July, 18 July, a nd 29 Aug. (Table 3-7). For the weeks in which the interaction was not significant, the main effect of cultivar was significant but the main effect of plant date was not. Cultivars differed significantly in their response to high temperatures. The interactions observed in 4 and 18 July were due to some cultivars having variable rating across plant dates while others had consis tent ratings. Serena White, Angelface White, and AngelMist Purple Stripe had variable ratings (Figure 3-2) due to the fact that PD 3 was continuing to increase in inflorescence number and PD 1 and 2 still had good flower coverage. In the other cultivars, however, the high temper atures had inhibited flowering and ratings were consistently low for all three plant dates. On 29 Aug., the deviation from the general trend was due to Serena White in PD 3 finally reaching it s peak flowering, which coincided with reduced

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76 flowering in the other two plant dates and no variab ility in other cultivars with respect to plant date. In general, cultivars were either heat tolerant or heat sensitive, regardless of plant date. Temperature throughout the season impacted the amount of flowering observed in each cultivar. AngelMist Purple Stripe had the most consiste nt flowering and never had ratings <3.0 after 18 July. AngelMist Purple Improved, in comparis on, did not have any ratings >2.0 all summer. The decrease in flowering during the summer months is most likely due to changes in temperature and not changes in DLI. M onthly DLI levels were 35, 37, and 30 molm-2d-1 for July, Aug., and Sept. respectively, and aver age temperatures were 26.8, 27.7, 25.7 C. A decrease in summer flowering and yield has been reported in canola at temperatures above 29.5 C (Morrison and Stewart, 2002) and at 33 C in tomato (Sato et al., 2000). Warner and Erwin (2005b) observed in Arabidopsis thaliana that floral buds began to abort at between 200 and 300 h of 33 C, and complete inflorescence failure occurred above 300 h of 33 C. In petunia, Kaczperski et al. (1991) observed flower abortion at temperatures above 25 C. In our study, many of the inflorescences formed flower buds but they failed to fully develop and open. On existing inflorescences, flowers opened sporadic ally along its length and were frequently separated by multiple sets of aborted buds. This decline in flowering is consistent with a decrease in net photosynthesis levels observed in angelonia at temper atures above 21 C, with AngelMist Purple Stripe having a higher optimal temperature than A ngelMist Deep Plum (Miller et al., 2001). This elevated net photosynthesis level may possibly indicate that it can tolerate higher temper atures better than other cultivars.

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77 Conclusions Angelonia a ngustifolia cultivars differed in their flower ing response to summer conditions. Plant height, inflorescence number, and flower size were affected by plant date x cultivar interactions. Inflorescence numbe r in all cultivars decreased w ith each successive planting, but the magnitude of the difference was cultivar dependent. For example, Serena White had a reduction in inflorescence number of 83 percent between PD 1 and 3 while AngelMist Purple Stripe had only a 53 percent decline. Plant date 1 had the highest peak floral display value and a longe r sustained length of flower display relative to the later plantings. Th e decline in flowering was primarily influenced by chronic high temperature stress during the summ er months and not plant age. For spring landscape plantings (April and early May) in North Florida, all cultivars will provide an acceptable amount of flower coverage. For plan tings after late May, only AngelMist Purple Stripe will likely provide acceptable flower coverage.

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78 0 2 4 6 8 10 5791113151719 Weeks after plantingFlower rating Plant date 1 Plant date 2 Plant date 3 minimum floweringA 0 2 4 6 8 10 5791113151719 Weeks after plantingFlower rating Plant date 1 Plant date 2 Plant date 3 minimum floweringE 0 2 4 6 8 10 5791113151719 Weeks after plantingFlower rating Plant date 1 Plant date 2 Plant date 3 minimum floweringB 0 2 4 6 8 10 5791113151719 Weeks after plantingFlower rating Plant date 1 Plant date 2 Plant date 3 minimum floweringF 0 2 4 6 8 10 5791113151719 Weeks after plantingFlower rating Plant date 1 Plant date 2 Plant date 3 minimum floweringC 0 2 4 6 8 10 5791113151719 Weeks after plantingFlower rating Plant date 1 Plant date 2 Plant date 3 minimum floweringG 0 2 4 6 8 10 5791113151719 Weeks after plantingFlower rating Plant date 1 Plant date 2 Plant date 3 minimum floweringD 0 2 4 6 8 10 5791113151719 Weeks after plantingFlower rating Plant date 1 Plant date 2 Plant date 3 minimum floweringH Figure 3-1. Flower ratings of Angelonia angustifolia graphed by weeks after transplant (WAT). The plant dates were 4 Apr., 2 May, a nd 30 May 2007, and the cultivars are A) AngelMist Dark Lavender B) AngelMi st Purple Improved C) AngelMist Lavender Stripe D) AngelMist Purple Stripe E) Serena White F) Angelina Violet and White G) Angelface Blue H) Angelface White.

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79 0 2 4 6 8 104 July18 July1 Aug.15 Aug.29 Aug.12 Sept.26 Sept.10 Oct. DateFlower rating Plant date 1 Plant date 2 Plant date 3 minimum floweringA 0 2 4 6 8 104 July18 July1 Aug.15 Aug.29 Aug.12 Sept.26 Sept.10 Oct. DateFlower rating Plant date 1 Plant date 2 Plant date 3 minimum flowering E 0 2 4 6 8 104 July18 July1 Aug.15 Aug.29 Aug.12 Sept.26 Sept.10 Oct. DateFlower rating Plant date 1 Plant date 2 Plant date 3 minimum flowering B 0 2 4 6 8 104 July18 July1 Aug.15 Aug.29 Aug.12 Sept.26 Sept.10 Oct. DateFlower rating Plant date 1 Plant date 2 Plant date 3 minimum flowering F 0 2 4 6 8 104 July18 July1 Aug.15 Aug.29 Aug.12 Sept.26 Sept.10 Oct. DateFlower rating Plant date 1 Plant date 2 Plant date 3 minimum flowering C 0 2 4 6 8 104 July18 July1 Aug.15 Aug.29 Aug.12 Sept.26 Sept.10 Oct. DateFlower rating Plant date 1 Plant date 2 Plant date 3 minimum floweringG 0 2 4 6 8 104 July18 July1 Aug.15 Aug.29 Aug.12 Sept.26 Sept.10 Oct. DateFlower rating Plant date 1 Plant date 2 Plant date 3 minimum floweringD 0 2 4 6 8 104 July18 July1 Aug.15 Aug.29 Aug.12 Sept.26 Sept.10 Oct. DateFlower rating Plant date 1 Plant date 2 Plant date 3 minimum floweringH Figure 3-2. Flower ratings of Angelonia angustifolia graphed by date of data collection. The plant dates were 4 Apr., 2 May, and 30 Ma y 2007. The cultivars are A) AngelMist Dark Lavender B) AngelMist Purple Impr oved C) AngelMist Lavender Stripe D) AngelMist Purple Stripe E) Serena Wh ite F) Angelina Violet and White G) Angelface Blue H) Angelface White.

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80 Table 3-1. Monthly temperat ure and DLI data for Gain esville, FL for summer 2007. Average daily maximum temperature (C) Average daily minimum temperature (C) Average 24-hr temperature (C) Average DLI (molm-2d-1) Apr 25.7 12.9 19.4 36.5 May 29.3 17.4 23.1 39.2 Jun 31.2 21.1 25.5 37.1 July 32.3 22.7 26.8 35.2 Aug 33.1 23.5 27.8 36.9 Sept 31.2 21.9 25.7 30.4 Oct (1-10) 29.1 20.5 24.1 25.8 Table 3-2. Split-plot analysis of variance for Angelonia angustifolia growth and flowering data collected at 10 weeks after planting for three plant dates. Significant levels for analysis of variance were P 0.05, 0.01, and 0.001. Plant height (cm) Number of inflorescences Flower number per inflorescence Flower height Plant date *** *** *** ** Cultivar *** *** *** *** Plant date x cultivar *** *** ***

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81Table 3-3. Mean separation for plant height (cm) of eight Angelonia angustifolia cultivars collected at 10 weeks after transplant for three plant dates. Uppercase letters indicate mean separation in each column and lowercase letters indicate mean separation in each row. Cultivar Plant date Serena White Angelina Violet and White AngelMist Dark Lavender AngelMist Purple Improved AngelMist Lavender Stripe AngelMist Purple Stripe Angelface White Angelface Blue 4 Apr 2007 (PD 1) 43.5 A c 56.0 AB bc 47.0 A c 49.2 AB c 54.2 A c 75.8 A a 77.0 A a 68.7 A ab 2 May 2007 (PD 2) 46.7 A e 63.2 A cd 51.0 A de 55.3 A de 64.3 A cd 81.7 A a 87.3 A a 69.5 A bc 30 May 2007 (PD 3) 32.7 A c 41.8 A bc 44.8 A bc 36.3 B c 56.3 A b 84.0 A a 74.5 A a 55.3 A b Table 3-4. Mean separation for number of inflorescences per plant of eight Angelonia angustifolia cultivars collected at 10 weeks after transplant for three plant dates. Uppercase letters in dicate mean separation in each column and lowercase letters indicate mean separation in each row. Cultivar Plant date Serena White Angelina Violet and White AngelMist Dark Lavender AngelMist Purple Improved AngelMist Lavender Stripe AngelMist Purple Stripe Angelface White Angelface Blue 4 Apr 2007 (PD 1) 171 A a 79 A cd 125 A b 25 A e 111 A bc 60 A de 22 A ef 11 A f 2 May 2007 (PD 2) 130 A a 11 B bc 26 B bc 3 A c 10 B bc 46 A b 17 A bc 5 A bc 30 May 2007 (PD 3) 30 B a 2 B a 13 B a 1 B a 9 B a 28 A a 5 A a 1 A a Table 3-5. Mean separation for flower height (mm) of eight Angelonia angustifolia cultivars collected at 10 weeks after transplant for three plant dates. Uppercase letters indicate mean separation in each column and lowercase letters indicate mean separation in each row. Cultivar Plant date Serena White Angelina Violet and White AngelMist Dark Lavender AngelMist Purple Improved AngelMist Lavender Stripe AngelMist Purple Stripe Angelface White Angelface Blue 4 April 2007 (PD 1) 17.2 A e 19.6 A cd 17.5 A de 21.5 A cd 19.5 A cde 22.3 A bc 25.0 A ab 25.6 A a 2 May 2007 (PD 2) 16.4 A b 15.9 A b 18.5 A b 16.6 B b 16.9 A b 21.9 A a 23.6 A a 23.3 A a 30 May 2007 (PD 3) 15.9 A d 19.6 A bcd 18.4 A d 17.9 AB cd 18.1 A cd 21.5 A b 25.1 A a 22.4 A ab

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82Table 3-6. Split-plot analysis of variance for Angelonia angustifolia flower ratings by plant age (weeks after transplant). 5 WATz 7 WAT 9 WAT 11 WAT 13 WAT 15 WAT 17 WAT 19 WAT Plant date ** *** *** *** ** ** Cultivar *** *** *** *** *** *** *** *** Plant date x cultivar *** *** *** *** *** *** NS z WAT = weeks after transplant Table 3-7. Split-plot analysis of variance for Angelonia angustifolia flower ratings by evaluation date. Week 27 Week 29 Week 31 Week 33 Week 35 Week 37 Week 39 Week 41 4 July 18 July 1 Aug. 15 Aug. 29 Aug. 12 Sept. 26 Sept. 10 Oct. Plant date *** NS NS NS *** NS NS NS Cultivar *** *** *** *** *** *** *** *** Plant date x cultivar *** *** NS NS NS NS NS

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83 CHAPTER 4 CULTIVAR BY FERTILIZER INTERA C TIONS AFFECT GROWTH OF Angelonia angustifolia Introduction Angelonia a ngustifolia is an herbaceous perennial native to the neotropics (USDA, 2006). Its popularity as a floriculture crop has risen in recent years due to virus indexing of unpatented plant material and to breeding efforts focused on more compact plants and an increased range of flower colors (Schoellhorn, 2002). Cultivars vary in growth habit, flower size and color, and flowering time (see Chapter 2). It is not known whether genotypes also vary in response to level of fertilization. Published production guidelines for angelonia vary by company. All recommend constant liquid feed with a complete fertilizer, but the level of fertilization ranges from N levels of 150 to 250 mgL-1 (Ball FloraPlant, 2006; PanAmerican Seed, 2006; Proven Winners, 2006; Fischer, 2008). Recommended N fe rtilizer guidelines published in the trade press range from 75 to 200 mgL-1 (Armitage, 1997; Schoellhorn and Alvar ez, 2002; Smith, 2007). It is unknown whether these differences in recommended levels are due to cultivar, series, or environmental conditions. Cultivar differences have been noted with poinsettia ( Euphorbia pulcherimma ), with optimal fertilization related to leaf color. Da rk-green leaved cultivars require an electrical conductivity (EC) of 1.5 to 2.0 mmhoscm-1 (mScm-1) using the saturated media extract procedure (SME), whereas medium-green l eaved cultivars require 2.0 to 2.5 mmhoscm-1 (Ecke, III et al., 2004). Temperature also influences fertilizer recommendations. The optimal fertilizer rate for petunia ( Petunia xhybrida ) decreases as temperature increases (Kang and van Iersel, 2001). It is unknown whether or not cultivars with in or across commercial angelonia series will respond similarly to changes in th e level of fertilization. The objectives of this experiment were

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84 to determine the response of si x angelonia cultivars to varying levels of fertilization and to determine the optimal concentration of fertilizer for angelonia production. Materials and Methods The six angelonia cu ltivars, Serena White AngelMist Dark Lavender, AngelMist Lavender Stripe, AngelMist Purp le Stripe, Angelface White and Angelface Blue, were selected for this experiment due to their range of growth habits and time to flower. See Expt. 2-2 for maintenance of stock plants and harvesting of unrooted cuttings. Forty-eight rooted cuttings of each cultivar were planted on 5 June 2007 into 2.2 L (16 cm diameter) pots filled with Fafard 2 soiless medium (Conrad Fafard, Inc., Apopka, FL), watered with regular water for 1 week. Plants were not fertilized to prevent a buildup of salts in th e medium before plants were transferred to subirrigation benches on 12 Ju ly. Treatments were N at 12.5, 25, 50, 100, 200, or 400 mgL-1using a 20.0N-4.4P-16.6K fertilizer (Peters 2010-20 Florida Special, The Scotts Co., Marysville, OH) (0.08, 0.16, 0.33, 0.65, 1.30, and 2.60 mSm-1) applied at each irrigation. The electrical conductivity (EC) of the irrigation water was 0.38 mScm-1. The experiment was set up as a split-plot design, with fertiliz er concentration as the main plot and cultivar as the sub-plot. One bench constituted a fertilizer treatment. W ithin each bench, cultivars were randomized in a complete random block design with four blocks a nd two plants of each cultivar per block. Plants were subirrigated as needed, every 2 to 3 days for the first 3 weeks and then daily for the remaining 3 weeks. Over the length of the e xperiment the average daily minimum, maximum, and 24-hour temperatures were 22.7, 36.2, and 28.0C, respectively. The average daily light level was 18 molm-2day-1. The number of days to first open flower we re recorded. SPAD readings and leachate pH and electrical conductivity (EC) were collected after 3 and 6 weeks. Plant height, plant width,

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85 number of inflorescences, and dr y weight (g) were collected af ter 6 weeks. SPAD readings indicate leaf greenness and we re taken using a SPAD 502 Minolta chlorophyll meter (Spectrum Technologies Inc., Plainfield, IL). Four readings per plant were recorded and averaged for both the upper leaves and the lower leaves. Readings for the upper leaves were taken on recently mature leaves and readings for the lower leaves were taken from the bottom 25% of the plant. Leachate pH and EC were collected using one pl ant per block for all cultivars on each bench. Leachate was collected 1 hour afte r irrigation using the PourThru method (Cavins et al., 2000). Leachate pH and EC were measured us ing a hand-held HANNA 98130 combination pH/EC meter (HANNA instruments, Woonsocket, RI). Pl ant height was measured from the top of the pot to the tallest growing tip. Plant width was measured at the widest point in the plant canopy and then again perpendicular the first measurement. Average plant width was used for analysis. Plant size was calculated as (plant height + average width)/2. Pl ants were harvested at the soil surface, bagged by block, and dried at 70 C for 48 hrs. Dry weight on a per plant basis was used for analysis. Data were analyzed usi ng SAS 9.1 (SAS Institute, Cary, NC) PROC MIXED and then regression analysis where appropriate. Results and Discussion Substrate pH and Electrical Conductivity The inte raction and the main effects of cultivar were not significant for pH, but the main effects of fertilizer concentra tion were significant at 6 weeks (Table 4-1). As a result, the regression equation for substrate pH relative to fertilizer c oncentration was pooled over all cultivars. As the fertilizer concentration incr eased, the general trend was a decrease in the pH value of the leachate. At 6 weeks, pH values ranged from 6.4 with N at 12.5 mgL-1 to 4.3 at 400 mgL-1 (Fig. 4-1). The6 week values were very si milar to the 3 week results, which ranged from 6.7 at 12.5 mgL-1 to 4.9 at 400 mgL-1. These results are consistent with the fact that Peters 20-

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86 10-20 (20N-4.4P-16.6K) has a potential acidity of 410 lbs CaCO3 equivalent per ton (The Scotts Company, 2004). Over time the pH of the medium and leachate will decrease, with the rate of decrease quicker as the fertilizer concentration applied is increased. The recommended pH range for angelonia is 5.5 to 6.2 (Ball, 2006). This range falls between the 25 and 50 mgL-1 treatments in this experiment. In this range, however, plants exhibited visual chlorotic symptoms. It is unkno wn why the soil solution pH fell below than the recommended range for plant growth in t hose treatments supplied with N at 100 mgL-1 and higher. The irrigation water used in this experiment had an alkalinity of 1.2-1.4 meq CaCO3, which should have provided some bufferi ng capacity against s udden drops in pH. The fertilizer x cultivar interaction was significant for leachate EC at 6 weeks. As the fertilizer concentration increased, the general tr end was an increase in leachate EC (Fig. 4-2), with the range between cultivars greater as fertilizer concentrati on increased. The EC range at the lowest and highest concentrations, 12.5 and 400 mgL-1, was 0.6 to 0.7 and 3.5 to 6.2 mScm-1, respectively. All cultivar s except Angelface White and AngelMist Purple Stripe exhibited a quadratic response (T able 4-2). These two cultiv ars, which exhibited a linear response, are the only two cultivars in this expe riment with thick, very pubescent, glandular leaves. The other cultivars ha d thinner, glabrous leaves. Whether or not these phenotypic differences are indicators of additional tolerance or sensitivity to fertilization is unknown since their growth responses were similar to the other cu ltivars in the range of fe rtilizer concentrations tested (see discussion below). The recommended EC range for angelonia is 0.6 to 0.9 mScm-1 using the 2:1 dilution method (Proven Winners, 2006), which is approx imately equivalent to 2.0 to 3.2 mScm-1 using

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87 the PourThru method (Cavins et al., 2000). Th is range falls between the 100 and 200 mgL-1 treatments, which is consistent with optim al plant growth and flowering (see below). Leaf Greenness SPAD readings, a nondestructive m easure of leaf greenness, are strongly correlated (greater than r2=0.9) with actual leaf ch lorophyll content obtained using destructive sampling (Uddling et al., 2007; Wang et al., 2005; Monje and Bugbee, 1992). A measure of the chlorophyll content in leaves is a strong indicator of th e N status of the plant. Although this experiment did not focus on the effects of increa sing N fertilization, but ra ther increasing levels of a complete fertilizer, the SPAD meter can be a useful tool for measuring the leaf greenness and the overall nutrient status of the plant. When single elements are withheld from an otherwise complete fertilizer, deficiency symptoms in angelonia for N, P, and K all appear at the same number of days after start of treatment (Williams, 2004). An indication of lower N content in the leaves, therefore, can help identify th e overall nutrient status of the plant. The fertilizer x cultivar interaction was highly significant at 3 and 6 weeks for both the upper and lower leaves (Table 4-1) The trend in all cu ltivars, in the upper and lower leaves, at both sampling intervals was an increase in th e SPAD value as the fertilizer concentration increased. However, the magnitude of the chan ge varied among cultivars. At 3 weeks, SPAD values in the lower leaves ranged from 36 to 41 at 12.5 mgL-1 and 52 to 59 at 400 mgL-1. Values in the upper leaves ranged from 41 to 51 and 54 to 59 at the lowest and highest concentrations, respectively (data not shown). Since the trends observed at 3 weeks are similar to those observed at 6 weeks, this discussion will be limited to the 6-week data. At 6 weeks, differences between cultivars were more pronounced. SPAD values in the lower leaves ranged from 15 to 26 at the lowe st concentration and 47 to 59 at the highest concentration (Fig. 4-3). Valu es in the upper leaves ranged fro m 31 to 43 and 54 to 59 at the

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88 lowest and highest concentrations, respectivel y (Fig. 4-4). In the lower leaves, Angelface White and AngelMist Purple Stripe had the lowest and highest SPAD values at 12.5 mgL-1, and Serena White and Angelface White had the lowest and highest values at 400 mgL-1. Serena White was the only cultivar with a linear response, with a ll others quadratic (Table 4-3). In the upper leaves, AngelMis t Lavender Stripe and Angelface White had the lowest and highest values at 12.5 mgL-1, and AngelMist Dark Lavender and AngelMist Lavender Stripe had the lowest and high est values at 400 mgL-1. Angelface White had the lowest SPAD value at 12.5 mgL-1 in the lower leaves, but also the highest value at 400 mgL-1 in the upper leaves, which may be an indication of increased nitrogen pa rtitioning or mobility in this cultivar relative to the others. There was little difference in the 6 week SPAD values for lower and upper leaves of plants in the 200 and 400 mgL-1 treatments. At 400 mgL-1, the ranges for the lower and upper leaves were 50 to 59 and 54 to 59, respectively. At 200 mgL-1, the ranges for the lower and upper leaves were 38 to 51 and 50 to 56, respectively. At 100 mgL-1 and below, the ranges no longer overlapped. The ranges for 100 mgL-1 were 32 to 42 for the lower leaves and 47 to 52 for the upper leaves. Even t hough the lower and upper leaf valu es no longer overlapped, the plants did not visually look chloroti c. At concentrations below 100 mgL-1, however, all cultivars at all concentr ations were visually chlorotic (F ig. 4-5). SPAD values in the upper leaves at 12.5 mgL-1, the lowest concentration used, were comparable to the lower leaf values recorded at 100 mgL-1. The lower leaves of the plants subjected to the 12.5 and 25 mgL-1 treatments exhibited symptoms of nutrient defi ciencies, including purpling of the leaves, light green or yellow coloration, and marginal necros is. As the fertilizer concentration became a

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89 limiting factor to plant growth, m obile nutrients were mobilized from the lower leaves to the upper leaves and meristems in order to support continued growth. Plant Growth The interaction was significant for height, widt h, size, and dry weight ind icating that the cultivars responded differently to ch anges in fertilizer levels. Tr ends for plant height and width are similar to those for plant size, so only the data for plant size will be presented. Plant size increased as the fertilizer concen tration increased up to maximum value, then decreased slightly (Fig. 4-6). Regression equations were significant for all cultivars except Angelface Blue and quadratic in nature (Tab le 4-4). Maximum plant size was obtained between 180 and 230 mgL-1. Serena White and Angelface White were the sma llest plants over the range of fertilizer concentrations and AngelMist Lave nder Stripe and AngelMist Purple Stripe were the largest. Both the two smallest and two largest cultivars ha d very different growth habits which inversely contributed to the determination of plant size. Of the two smallest cultivars, Serena White is very short and compact in habit, while Angelfac e White is naturally tall and narrow with very little lateral branching. Of the two largest cultiv ars, AngelMist Lavender Stripe is the shorter cultivar but has more of a spr eading habit than Serena White, while AngelMist Purple Stripe is the tallest cultivar in this experiment and de velops lateral branches shortly after the start of flowering (for additional descriptions of cultivars, see Chapter 2). Changes in plant size were due to changes in bo th plant height and width. As fertilizer concentrations increase from suboptimal to optim al, a small increase in concentration will have a large positive effect on plant grow th. As fertilizer concentra tions continue to increase beyond what is required for normal plant growth, elevat ed soluble salts levels tend to occur in the substrate, especially in subirrig ated containers where leaching does not occur. This increase, as

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90 monitored by EC levels, can result in decreased plant grow th (Whipker et al., 2003). The optimal fertilizer concentration for plant size in this experiment was around 200 mgL-1 for most cultivars. The interaction for plant dry weight was si gnificant. For those cultivars that had a quadratic response to increased fertilization, dry weight (g ) per plant increased as the concentration applied in creased up to a maximum between 232 and 272 g, and then decreased at higher concentrations (Fig. 4-7). In AngelMis t Lavender Stripe, the response was linear and did not reach a plateau at the concentrations te sted. Angelface Blue did not have a significant regression equation, and the highest observed value was at 100 mgL-1. In general, cultivars that were larger in plant size tende d to have higher dry weights. AngelMist Lavender Stripe and AngelMist Purple Stripe had the highest dry weights over al l concentrations except at 12.5 mgL-1, and AngelMist Dark Lavender, Angelface Wh ite, and Serena White had the lowest. As the fertilizer concentration increased from a de ficient level to an optimal level, the plant dry weight increased. Above an optimal level, the hi gh EC levels in the leach ate inhibited growth to some degree and plants accumulated le ss dry weight on a per plant basis. Flowering The cultivar x f ertilizer interaction for number of inflorescences per plant was significant. Large differences were observed in the responses of the cultivars to increased fertilization. The general trend was an increase in the number of inflorescences as fertilizer concentration increased. Between 200 and 400 mgL-1, inflorescence number either declined slightly or remained unchanged (Fig. 4-8). Angelface White and Serena White had the least and greatest number of inflorescences, respectively, at all con centrations. The three cultivars with the lowest number of inflorescences Angelface White, A ngelface Blue, and AngelMist Purple Stripe are cultivars that tend to have fewer inflorescences per plant and a greater number of flowers

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91 per inflorescence (see Chapter 2). The interacti on for number of days to first flower was not significant. The differences observed were due to the main effects of cultivar, but not fertilizer concentration (Table 4-5). The average number of days to first flower in this experiment was very similar to the results of a cultivar screen conducted one mont h earlier in the same greenhouse with these cultivars (see Chapter 2). Seren a White flowered in an average of 22 d and was the earliest to flower relative to all other cultivars except for AngelMist Da rk Lavender. Angelface White was the latest to flower and averaged 43 d. It was almost 2 weeks later than AngelMist Purple Stripe and nearly 3 weeks later than the othe r cultivars. Flowering was not significantly enhanced or delayed by fertilizer concentra tion. These results ar e in contrast to Salvia splendens in which flowering was delayed by 8 d at lower fertilizer concentrations (Kang and van Iersal, 2004). Thus, the length of the crop cycle in ange lonia will be unaffected if slight nutritional deficiencies are induced as a means to cont rol crop vegetative gr owth during production. Conclusions There was n ot a particular fertilizer concen tration that yielded maximum growth and flowering for all parameters. The response of cu ltivars to increased ferti lizer concentration was not cultivar-specific, indicating that they will respond similarly to changes in the concentration applied. Leaf greenness, a primary visual indicator of the nutritiona l status of a plant, increased in both the upper and lower leaves as the fertil izer concentration app lied increased. Maximum plant size occurred between 180 and 230 mgL-1, maximum plant dry weight (g) occurred between 230 and 275 mgL-1, and maximum inflorescence number occurred between 245 and 340 mgL-1. The recommended EC range (2.0 to 3.2 mScm-1) fell between the 100 and 200 mgL-1 treatments.

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92 The selection of a fertilizer concentrati on is based upon many factors, including the concentration required for acceptable plant qualit y, the irrigation method used, and the cost of the fertilizer. Best management practices r ecommend using the least amount of fertilizer necessary to ensure adequate growth and development of th e crop. Although maximum plant growth occurred at concentra tions greater than 100 mgL-1, it was the lowest fertilizer concentration at which all cultivars were not visually chlorotic. Plant growth and quality were acceptable for the retail market. Recommendations from this experiment would be to use a fertilizer concen tration of 100 mgL-1 or slightly higher for the warm-season production of subirrigated angelonia, regardless of cultivar. Slight adjustments may need to be made for climate and time of year. Previous recommendations for angelonia fe rtilization ranged from N at 75 to 200 mgL-1. Differences in recommendations were not defined as being specific to any cultivar or irrigation method. Typically, fertilizer recommendations are based upon overhead irrigation. Since no leaching occurs in subirrigation systems, it is generally recommended that growers use a halfstrength fertilizer solution when using subirr igation instead of overhead irrigation (Nelson, 1994). The results from this study agree with pr oduction guidelines of overhead irrigating with 150 to 200 mgL-1 as published for these cultivars by their suppliers a nd by Schoellhorn and Alvarez (2002). However, this range is much narrower than what had been published in the trade press.

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93 4.0 4.5 5.0 5.5 6.0 6.5 7.0 050100150200250300350400 Fertilizer N concentration (mgL-1)pH Figure 4-1. Effect of incr easing fertilization with 20.0N4.4P-16.6K on substrate pH in Angelonia angustifolia 6 weeks after start of treatments. Treatment N concentrations were 12.5, 25, 50, 100, 200, or 400 mgL-1. The regression line was generated from individual plant data and the points on th e graph are the means for each concentration pooled over cultivar (n= 24). The equation and r2 value can be found in Table 4-2. 0 1 2 3 4 5 6 7 050100150200250300350400 Fertilizer N concentration (mgL-1)Electrical conductivity (mScm-1) Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue Figure 4-2. Effect of increas ing fertilization with 20.0N-4.4P -16.6K on substrate electrical conductivity in Angelonia angustifolia 6 weeks after start of treatments. Treatment N concentrations were 12.5, 25, 50, 100, 200, or 400 mgL-1. The regression lines were generated from individual plant data and the points on the graph are the means for each individual treatment (n=8). Regression equations and r2 values can be found in Table 4-2.

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94 10 20 30 40 50 60 050100150200250300350400 Fertilizer N concentration (mgL-1)SPAD Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue Figure 4-3. Effect of increas ing fertilization with 20.0N-4.4P16.6K on lower leaf SPAD values in Angelonia angustifolia 6 weeks after start of treatments. Treatment N concentrations were 12.5, 25, 50, 100, 200, or 400 mgL-1. SPAD units indicate relative leaf greenness. The regression lines were genera ted from individual plant data and the points on the graph are the m eans for each individual treatment (n=8). Regression equations and r2 values can be found in Table 4-3. 30 35 40 45 50 55 60 050100150200250300350400 Fertilizer N concentration (mgL-1)SPAD Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue Figure 4-4. Effect of increas ing fertilization with 20.0N-4.4P16.6K on upper leaf SPAD values in Angelonia angustifolia 6 weeks after start of treatments. Treatment N concentrations were 12.5, 25, 50, 100, 200, or 400 mgL-1. SPAD units indicate relative leaf greenness. The regression lines were genera ted from individual plant data and the points on the graph are the m eans for each individual treatment (n=8). Regression equations and r2 values can be found in Table 4-3.

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95 A B C D E F Figure 4-5. Response of Angelonia angustifolia AngelMist Dark Lavender to increasing rates of fertilization with 20.0N-4.4P-16.6K. N concentrations listed are in mgL-1. A) 12.5 B) 25 C) 50 D) 100 E) 200 F) 400.

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96 30 40 50 60 70 050100150200250300350400 Fertilizer N concentration (mgL-1)Size (cm) Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue Figure 4-6. Effect of incr easing fertilization with 20.0N -4.4P-16.6K on plant size in Angelonia angustifolia 6 weeks after start of treatments. Treatment N concentrations were 12.5, 25, 50, 100, 200, or 400 mgL-1. Plant size calculated as [(plant height + average plant width)/2]. The regression lines were generated from individual plant data and the points on the graph are the means for each individual treatment (n=8). Regression equations and r2 values can be found in Table 4-4. 0 10 20 30 40 50 60 70 050100150200250300350400 Fertilizer N concentration (mgL-1)Dry weight (g) Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue Figure 4-7. Effect of increas ing fertilization with 20.0N-4.4P16.6K on plant dry weight in Angelonia angustifolia 6 weeks after start of treatments. Treatment N concentrations were 12.5, 25, 50, 100, 200, or 400 mgL-1. The regression lines were generated from individual plant data and the points on the graph are the means for each individual treatment (n=8). Regression equations and r2 values can be found in Table 4-4.

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97 0 10 20 30 40 50 60 70 80 90 100 050100150200250300350400 Fertilizer N concentration (mgL-1)Inflorescence numbe r Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue Ser White AM Dk Lav AM Lav Stripe AM Purp Stripe AF White AF Blue Figure 4-8. Effect of increasing fert ilization with 20.0N-4.4P-16.6K on number of inflorescences in Angelonia angustifolia 6 weeks after start of treatments. Treatment N concentrations were 12.5, 25, 50, 100, 200, or 400 mgL-1. The regression lines were generated from individual plant data and the points on the graph are the means for each individual treatment (n=8). Regression equations and r2 values can be found in Table 4-4.

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98Table 4-1. Analysis of variance for Angelonia angustifolia cultivars in respons e to fertilization 3 wk SPAD upper leaves 3 wk SPAD lower leaves 6 wk SPAD upper leaves 6 wk SPAD lower leaves 3 wk pH 6 wk pH 3 wk EC 6 wk EC Plant height (cm) Plant width (cm) Plant size (cm) Dry weight (g) Inflor. Number Days to flower Fertilizer *** *** *** *** *** *** *** *** *** *** *** *** *** NS Cultivar *** *** ** *** ** NS NS *** *** *** *** *** *** Fert x Cv *** *** *** *** *** NS NS *** *** *** *** *** NS NS, *, **, *** nonsignificant and significant at P 0.05, 0.01, and 0.001, respectively. Table 4-2. Regression equa tions for pH and EC measurements co llected at 6 weeks after treatment. Variable Cultivarz Significancey r2 Equation pH Q*** 0.73 y = 6.4 0.01x + 0.00002x2 EC (mScm-1) SW Q* 0.76 y = 0.1 + 0.02x 0.00003x2 AMDL Q* 0.66 y = 0.1 + 0.03x 0.00004x2 AMLS Q* 0.94 y = 0.6 + 0.001x + 0.00001x2 AMPS L*** 0.93 y = 0.5 + 0.001x AFW L*** 0.86 y = 0.6 + 0.01x AFB Q* 0.81 y = 0.3 + 0.02x 0.00003x2 z Cultivar abbreviations: SW Serena Wh ite, AMDL AngelMist Dark Lavender, AMLS AngelMist Lavender Stripe, AMPS AngelMist Purple Stripe, AFW Angelface White, AFB Angelface Blue. y *, **, *** significant at P 0.05, 0.01, and 0.001, respectively.

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99Table 4-3. Regression equations for SPAD readings collected at 6 weeks after treatmentz. Variable Cultivarz Significancey r2 Equation 6 week SPAD SW Q** 0.58 y = 41.7 + 0.08x 0.0001x2 (upper leaves) AMDL Q* 0.37 y = 45.3 + 0.07x 0.0001x2 AMLS Q*** 0.66 y = 35.7 + 0.15x 0.0002x2 AMPS Q*** 0.57 y = 39.9 + 0.14x 0.0003x2 AFW Q* 0.54 y = 44.8 + 0.07x 0.0001x2 AFB Q* 0.67 y = 39.0 + 0.08x 0.0001x2 6 week SPAD SW Q*** 0.76 y = 22.3 + 0.16x 0.0002x2 (lower leaves) AMDL L*** 0.32 y = 47.4 + 0.02x AMLS Q*** 0.83 y = 20.6 + 0.23x 0.0004x2 AMPS Q*** 0.85 y = 23.2 + 0.19x 0.0003x2 AFW Q*** 0.92 y = 13.1 + 0.20x 0.0002x2 AFB Q*** 0.87 y = 25.4 + 0.16x 0.0002x2 z Cultivar abbreviations: SW Serena Wh ite, AMDL AngelMist Dark Lavender, AMLS AngelMist Lavender Stripe, AMPS AngelMist Purple Stripe, AFW Angelface White, AFB Angelface Blue. y *, **, *** significant at P 0.05, 0.01, and 0.001, respectively.

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100Table 4-4. Regression equations for plan t size, dry weight, and number of inflorescences at 6 weeks after treatment. Variable Cultivarz Significancey r2 Equation Plant size (cm) SW Q*** 0.33 y = 41.5 + 0.09x 0.0002x2 AMDL Q*** 0.36 y = 47.3 +0.09x 0.0002x2 AMLS Q* 0.14 y = 59.5 + 0.04x 0.0001x2 AMPS Q*** 0.36 y = 55.0 + 0.14x 0.0003x2 AFW Q* 0.14 y = 47.4 + 0.05x 0.0001x2 AFB NS Dry weight (g) SW Q* 0.43 y = 13.0 + 0.15x 0.0003x2 AMDL Q* 0.27 y = 15.4 + 0.12x 0.0002x2 AMLS L* 0.26 y = 32.2 + 0.08x AMPS Q*** 0.52 y = 9.8 + 0.50x 0.001x2 AFW Q*** 0.58 y = 18.4 + 0.16x 0.0003x2 AFB NS Inflorescence Number SW Q*** 0.67 y = 23.4 + 0.57x 0.001x2 AMDL L** 0.20 y = 35.4 + 0.07x AMLS Q*** 0.58 y = 20.3 + 0.19x 0.0003x2 AMPS Q* 0.20 y = 4.0 + 0.04x 0.00008x2 AFW L*** 0.41 y = 0.6 + 0.006x AFB Q* 0.46 y = 4.6 + 0.05x 0.00007x2 z Cultivar abbreviations: SW Serena Wh ite, AMDL AngelMist Dark Lavender, AMLS AngelMist Lavender Stripe, AMPS AngelMist Purple Stripe, AFW Angelface White, AFB Angelface Blue. y NS, *, **, *** nonsignificant and significant at P 0.05, 0.01, and 0.001, respectively.

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101 Table 4-5. Number of days to first open flower in Angelonia angustifolia The fertilizer x cultivar interaction and th e main effect of cultivar were not significant at P 0.05. The main effect of cultivar displayed in th is table are pooled over all fertilizer concentrations for each cultivar. Mean separation was performed using Tuleys at =0.05. Cultivarz Number of days Mean separationx AFW 43.0 a AMPS 29.9 b AMLS 26.9 c AFB 25.5 cd AMDL 23.8 de SW 22.2 e z Cultivar abbreviations: SW Serena White, AMDL AngelMist Dark Lavender, AMLS AngelMist Lavender Stripe, AMPS AngelMist Purple Stripe, AFW Angelface White, AFB Angelface Blue

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102 CHAPTER 5 Angelonia angustifolia CULTIVARS DIFFER I N RESPONSE TO PACLOBUTRAZOL, DAMINOZIDE, AND ETHEPHON Introduction Comm ercially-available Angelonia angustifolia cultivars differ significantly in growth habit (Chapter 2), and many tend to become ex cessive in height duri ng production without the use of plant growth regulato rs (PGRs). Commonly used PG Rs on bedding plants include chlormequat chloride, daminozide, ethephon, p aclobutrazol, and uniconazole. With the exception of ethephon, all inhibit cell elongation by blocking gibberellin synthesis (Gent and McAvoy, 2002). Ethephon releases ethylene within plant tissues, which ca n result in shorter internodes and increased lateral branching (Beau dry and Kays, 1988). Plant response to PGR applications varies by chemical and concentration applied (Ba rrett, 2001), method of application (Gibson and Whipker, 2003), and species or cult ivar sensitivity (Gibson and Whipker, 2001a; Lewis et al., 2004; Starman et al., 2004; Warner and Erwin, 2003). Cultivar differences in response to PGR applications are often due to di fferences in sensitivity to the PGR applied, and this may or may not correlate with plant vigor (Ecke III et al., 2004). Research indicates that at least some ange lonia cultivars are responsive to daminozide, ethephon, and paclobutrazol applic ations. Blue Pacific pl ants treated with 5000 mgL-1 daminozide were shorter than untreated plants but paclobutrazol sprays of up to 80 mgL-1 were ineffective at controlling height (Starma n, 2001). Miller and Armitage (2002), however, observed that five cultivars in the AngelMist series had simila r responses to PGRs and that paclobutrazol sprays at 50 and 100 mgL-1, as well as ancymidol at 50 and 100 mgL-1 and daminozide at 2500 and 5000 mgL-1 effectively controlled plant he ight. It is unknown if current cultivars and/or series will vary in their sensitivity to the type and concentration of PGR applied.

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103 Published recommendations for PGR applications for growth control of angelonia suggest spraying daminozide at 2500 mgL-1, uniconazole at 2 to 5 mgL-1; paclobutrazol at 5 mgL-1; or a tank mix of chlormequat chloride plus daminozide at 700 to 1000 and 1500 to 2000 mgL-1, respectively (Ball FloraPlant, 2006; PanAmerican Seed, 2006; Proven Winners, 2006; Selecta, 2008). Proven Winners recomme nds avoiding the use of ethephon on cultivars in the Angelface series due to its ability to cau se leaf tip burn or distorted growth, whereas the other companies neither recommend using or avoiding this chemi cal. It is unknown whether the phytotoxicity symptoms observed in the Angelf ace series are series-specific. The objectives of these experiments were 1) to determine which PGR chemicals and which concentrations will effectively control growth of angelonia, and 2) whether or not response to the PGRs is cultivar specific or not. Four experime nts were conducted: a ra te screen examining the efficacy of daminozide, ethephon, and paclobutrazol on seven cultivars, and three more in-depth studies, each focusing solely on daminozide, pacl obutrazol, or ethephon in order to look at differences between representative cultiv ars under different growing conditions. Materials and Methods Experiment 5-1 Paclobutrazol, Damino z ide, and Ethephon Response Curves Cuttings from seven Angelonia angustifolia cultivars Angelina Violet and White, AngelMist Dark Lavender, AngelMist Purp le Improved, AngelMist Lavender Stripe, AngelMist Purple Stripe, Angelface White, and Angelface Blue were harvested from stock plants on 14 May 2007 (see Expt. 2-2 for stock plant culture). Sixty rooted cuttings of each cultivar were planted on 12 June, into 2.2 L (16 cm diameter) pots filled with Fafard 2 soilless medium (Conrad Fafard, Inc., Apopka, FL) and pinched to two nodes. Two weeks later, on 26 June, the following plant growth regulator (PGR) treatments were applied: paclobutrazol (Bonzi; Syngenta Corp., Greensboro, NC) drenches at 5, 10, or 20 mgL-1; ethephon (Florel;

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104 Monterey, Fresno, CA) sprays at 250, 500, or 1000 mgL-1 plus Capsil (Aquatrols Corp. of America; Paulsboro, NJ) at 0.5mLL-1; daminozide (B-Nine; OHP, Mainland, PA) sprays at 1250, 2500, or 5000 mgL-1; and an untreated control. Drench volume was 180 ml per pot for paclobutrazol, and spray volumes were 306 mLm-2 for ethephon and 204 mLm-2 for daminozide, respectively.. Plants were grown in a naturally ventilated greenhouse and continuously liquid-fed as needed with N at 150 mgL-1 using 20.0N-4.4P-16.6K (Peters 20-1020 Florida Special, The Scotts Co., Marysville, OH). Average daily minimum, maximum, and 24-hour temperatures over the length of th e experiment were 22.7, 35.9, and 27.9 C, and the average daily light integral (DLI) was 17.8 molm-2d-1. Plant height, measured from the rim of the pot to the tallest point on the plant, was measured at treatment and again 23 d later, on 19 July, once plants reached a marketable stage. Stem elongation was calculated as the difference be tween the two heights. The experiment with 10 PGR treatments and seven cultivars was set up in a split plot design, with PGR as the main plot and cultivar as the sub-plot. Plants we re randomized in a complete random block design with three blocks and two plants of each cultivar per block. Data were analyzed by regression analysis in SAS 9.1 (SAS Institute, Cary, NC) in order to generate response curves for each cultivar. Each PGR was analy zed individually, with a common unt reated control used for each cultivar for all three growth regulators. Experiment 5-2 Daminozide The cultivars AngelMis t Dark Lavender, AngelMist Purple St ripe, and Angelface White were selected from those used in Expt. 51 because they were examples of a wide range of growth habits. See Expt. 5-1 for descripti on of materials and methods Forty-eight rooted cuttings of each cultivar were transplanted in to 2.2 L (16 cm diameter) pots on 11 July 2007. Daminozide (B-nine; OHP, Mainland, PA) sprays were applied on 27 July, at 0, 1250, 2500, or

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105 5000 mgL-1 at a spray volume of 204 mLm-2. Average daily minimum, maximum, and 24-hour temperatures over the length of the experime nt were 23.8, 37.5, and 29.1 C, and average DLI was 18.8 molm-2day-1. Final height was measured on 22 Aug. and stem elongation was calculated. The experiment was set up as a fact orial with four daminozide concentrations and three cultivars. Plants were randomized in a complete random block design with three blocks and four plants of each treatment per block. An alysis of variance and Tukeys mean separation at =0.05 were conducted using SAS. Experiment 5-3 Paclobutrazol Spray and Drench Applications See Expt. 5-1 for m aterials and methods. Un rooted cuttings from AngelMist Purple Stripe and Angelface White were harvested on 25 July 2007, and 81 rooted cuttings of each cultivar were transplanted into 2.2 L (16 cm diameter) pots on 17 Aug. Paclobutrazol sprays were applied 2 weeks after pinch at 50 or 100 mgL-1 at a volume of 204 mLm-2. For drench applications, paclobutrazol concentrations were 2, 4, or 8 mgL-1 using a drench volume of 180 mL per pot. Applications were made at visi ble bud, which was 4 and 6.5 weeks after pinch for AngelMist Purple Stripe and Angelface White respectively. Average daily minimum, maximum, and 24-hour temperatures over the le ngth of the experiment were 22.7, 34.5, and 27.1 C, and average DLI was 14.8 molm-2day-1. Final height was measured when control plants reached a marketable stage, on 28 Sept. and 15 Oct. for AngelMist Purple Stripe and Angelface White, respectively, and stem elongation was calculated. Plants were arranged in a randomized complete block design with 3 blocks and 3 plants of each treatment per block. Analysis of variance and m ean separation using Tukeys =0.05 were conducted using SAS. Experiment 5-4 Ethephon Rooted liners of AngelMist Dark L avender, AngelMist Purple Improved, and AngelMist Purple Stripe from Ball FloraPla nt (West Chicago, IL) and rooted liners of

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106 Angelface White Improved from EuroAmerican Propagators (Bonsall, CA) were received the week of 1 Oct. 2007. Forty-eight plants of each cultivar were planted on 8 Oct. into 650 cm3 (11 cm) pots filled with Fafard 2-P soilless medium (Conrad Fafard, Inc.; Apopka, FL) and pinched to two nodes. Ethephon applications were a pplied 10 d after transplant, on 18 Oct., at concentrations of 0, 250, 500, or 1000 mgL-1 using techniques given in Expt. 5-1. This experiment was designed as a factorial co nsisting of four cultivars and four ethephon concentrations. Plants were arranged in a random ized complete block design, with three blocks, 16 treatments, and three plants per treatment pe r block. They were grown on a subirrigation bench and fertilized with N at 75 mgL-1 using 20.0N-4.4P-16.6K (Peters 20-10-20 Florida Special, The Scotts Co., Marysv ille, OH). Average daily maximum, minimum, and 24-hour temperatures and DLI during the first week of the experiment were 31.5, 19.5, and 24.6 C and 9.8 molm-2day-1, and during the last week were 21.2, 13.4, and 16.7 C and 7.8 molm-2day-1, respectively. At 1 week after treatment (1 WAT), plan ts were visually ra ted for appearance of phytotoxicity symptoms on a 0 3 scale, with 0 = no visible symptoms, 1 = slight chlorosis of leaf tips, 2 = moderate chlorosis of leaves and slight necrosis at leaf edges, and 3 = severe chlorosis of leaves, moderate to severe leaf necrosis, and leaf distor tion. Plant height was measured at 4 WAT and stem elongation was calcula ted. The number of laterals per plant was determined at 5 WAT. All shoots greater than 5 cm were considered as laterals, and no distinction was made as to whether they were primary or secondary laterals. Plants were observed daily and date of fi rst flower was recorded. Analysis of variance was determined for all parameters using SAS. Regression analysis was run by cultivar on plant height, width, and si ze, and number of days to flower. Mean

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107 separation using Tukeys at =0.05 was determined for phytotoxicity rating and number of laterals per plant. Results and Discussion Experiment 5-1 The chem ical x cultivar interaction wa s significant. Daminozide, ethephon, and paclobutrazol were effective at co ntrolling plant growth, but their effectiveness varied with the cultivar and concentration applied. For each PG R, the concentrations selected were based upon published recommendations. The middle con centration was the one recommended for production, and the higher and lower concentrations were selected in order to generate response curves. Paclobutrazol All cultivars exhibited a significant quadrati c response to paclobutrazol, with a general decrease in stem elongation as the concentration increased (Table 5-1, Fig. 5-1a). All of the regression curves showed a strong linear component between the untreated control plants and the 5 mgL-1 concentration, and as th e concentration applied in creased from 5 to 20 mgL-1, there was a smaller incremental reduction in stem elongation. Cultivars varied in their respons e to paclobutrazol. At 5 mgL-1, stem elongation in treated plants was 22 to 65 per cent of untreated controls. A ngelina Violet and White was the most sensitive cultivar and AngelMist Purple Improved and AngelMist Dark Lavender were the least sensitive cultivars. The percent stem elongation observed in Angelina Violet and White at 5 mgL-1 was comparable to Angelface Blue or AngelMist Purple Improved plants treated with 20 mgL-1 (data not shown). Paclobutrazol drenches at 5 mgL-1 have been recommended for height control of angelonia cultivars grown in warm climates (PanAmerican Seed, 2006). However, 5 mgL-1 was

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108 too high for most cultivars, even though applic ations were applied du ring the summer. In AngelMist Purple Stripe, a vigorous cultivar, the efficacy of the application had started to decrease by the time data was co llected and the inter nodes in the inflorescen ce were beginning to elongate almost normally (Figure 5-2). Concentrations less than 5 mgL-1 should be trialed on an individual cultivar basis for a more precise determination of optimal drench concentration. Daminozide Stem elongation decreased as the concentration applied increased in all cultivars (Fig. 51b, Table 5-1). Angelina Violet and White an d AngelMist Purple Stripe had significant quadratic responses and the other five cultiv ars exhibited linear re sponses to increasing daminozide concentration. Stem elonga tion in plants sprayed with 5000 mgL-1 was 62 to 87 percent of the untreated contro ls. Angelina Violet and White and Angelface Blue were the two most responsive cultivars. AngelMist Da rk Lavender was the least responsive cultivar. AngelMist Purple Stripe and Angelface Blue had the greatest stem elongation at all concentrations applied. Both of these cultiv ars are tall plants and naturally have longer internodes than more compact cultivar s. At concentrations of 2500 mgL-1 and below, both cultivars had similar responses to daminozide. However, at 5000 mgL-1, Angelface Blue had a greater response to the a pplication than AngelMist Purple Stripe(Fig. 5-1b). The recommended spray concentration for daminozide is 2500 mgL-1 (Selecta, 2008), and it is applied at much higher concentrations than paclobutrazol because it has much lower activity (Barrett, 2001). It is commonly used on bedding crops in a tank mix solution with chlormequat chloride. The concentration of daminozide us ed varies depending upon the concentration of chlormequat chloride used, but ranges from 1500 to 2500 mgL-1. Daminozide applied by itself is not as effective as using a tank mix, but can be effective in certain crops.

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109 The cultivars in this experiment varied in their response to daminozide, from moderately effective to ineffective (Fig. 5-3). This is simila r to the varied response observed in a trial of 26 ornamental cabbage and kale ( Brassica oleracea ) cultivars in which efficacy was observed in two cultivars when sp rayed with 2500 mgL-1 and in eight cultivars when sprayed with 5000 mgL-1 (Gibson and Whipker, 2001a). Ethephon The general trend observed with ethephon wa s that stem elongation decreased as the concentration applied increased (F ig. 5-2c, Table 5-1). All of the cultivars had significant linear regression responses. AngelMist Purple Stri pe was the most responsive cultivar and AngelMist Dark Lavender was the least responsi ve cultivar. Stem elongation in plants treated with 1000 mgL-1 ranged from 54 to 80 percent of untreated plants, respectively. The efficacy of ethephon in controlling hei ght of vegetative annuals is both crop and cultivar specific. Hammond et al. (2007) observed that blanketflower ( Gaillardia pulchella ) Torch Flame was more compact than untreat ed plants but a native Florida ecotype was insensitive to ethephon applicati ons. Starman et al. (2004) obse rved in a wide selection of vegetative annuals that 81% were responsive to ethephon applications. Multiple cultivars were trialed within seven species and cul tivar differences were observed in Antirrhinum majus Calibrachoa hybrids, and Petunia xhybrida, but not in Diascia xhybrida Impatiens wallerana Lantana camara, or Nemesia xhybrida. Experiment 5-2 The cultivar x dam inozide concentration inte raction was not significant, but the main effects of cultivar and concentration were signifi cant. Within each concentration, the averages were pooled across cultivars, and within each cultivar, the averages were pooled across concentrations for mean separation (Table 5-2).

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110 For the main effects of concentration, all co ncentrations were signi ficantly different from the untreated control and a ra te response was observed. Stem elongation was not significantly different between 1250 and 2500 mgL-1, but 5000 mgL-1 was significantly different from the other treatments. These results ar e similar to those observed in Expt 5-1, in which an increase in concentration decreased stem elongation. All three cultivars had similar responses to daminozide, even though they had different growth habits. In Expt. 5-1, stem el ongation in plants sprayed with 5000 mgL-1 was 73 to 82 percent of untreated plants, de pending upon cultivar, and in Expt 5-2, stem elongation was 78 to 87 percent of untreated plants. Plants were slight ly less responsive in Expt. 5-2 due to the higher average daily temperatures under which the plants were grown. Angelfac e White was the most responsive cultivar to daminozide in both experiments. AngelMist Dark Lavender had the greatest decrease in sensitivity between the two experiments, in which plants treated with 5000 mgL-1 were 75 percent the height of untreated plan ts in Expt. 5-1 but 87 percent in Expt. 5-2. For the main effects of cultivar, all three cultivars were significantly different from each other. These differences can be explained ba sed upon each cultivars natural growth habit (see Chapter 2). Plant height, from tallest to shortest, was AngelMist Purple Stripe, Angelface White, and AngelMist Dark Lavender, resp ectively. AngelMist Purple Stripe and Angelface White are both tall, vi gorous cultivars and AngelMist Dark Lavender is a short, compact cultivar. Daminozide applications were effective at controlling plant height, but a high concentration is needed for efficacy. In warm climates and during the summer months, it is not the most effective PGR for this crop. However, in the cooler months and in northern states, it

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111 may be a viable option since angelonia cultivars are more responsive to daminozide than paclobutrazol sprays. Experiment 5-3 The cultivar x trea tment interaction (with treatment defined as the drench and spray applications at varying concentr ations) and the main effect of paclobutrazol treatment, were not significant, but the effect of cultivar was significa nt (Table 5-3). AngelMist Purple Stripe is a slightly taller cultivar than A ngelface White at marketability in the late summer. None of the treatments resulted in pl ants with less stem elongation than untreated plants. The paclobutrazol spray applied 2 weeks after pinch was not effective at controlling plant height, which is consistent with results from Starman (2001) but inconsistent with results observed by Miller and Armitage (2002). The la ter drenches applied at visible bud in this experiment were not effective at controlling plant height either, but most likely because they were applied too late in producti on to significantly alter final height. Both of these cultivars are tall and have vigorous growth habits and will need to receive a paclobutrazol drench, rather than a spray, within the first 2 to 3 weeks after pinch in order to obtain adequate height control. AngelMist Purple Stripe was treated 4 weeks af ter pinch, and by then it had already grown too tall. Paclobutrazol drenches at 5 mgL-1 have been recommended for height control of angelonia cultivars grown in warm climates (P anAmerican Seed, 2006). Uniconazole, another triazole PGR, has been recommended as a drench application at 2 to 5 mgL-1 (Proven Winners, 2006). Uniconazole is more active than paclobu trazol and in gene ral, paclobutrazol concentrations are required to be two to four tim es higher than uniconazole in order to get the same degree of efficacy (Barrett, 2001).

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112 Based on these recommendations, dren ch applications of 5 to 10 mgL-1 paclobutrazol should have resulted in marketable plants but instead resulted in severely stunted plants. Early sprays, which have been shown to be effective at 50 and 100 mgL-1 (Miller and Armitage, 2002), and late drenches at lower concentrations were used in E xpt. 5-3 in order to examine if the timing of the drench applica tion affects efficacy and if early sprays would be a viable option instead of drench application. However, these cultivars did not respond to the spray or drench concentrations used in Expt. 5-3. Spray appl ications will not be practical due to the high concentration required for efficacy. Drench appli cations will need to be applied carefully and concentrations will need to be altered dependi ng upon the age of the plant. Early in production, lower concentrations should be effective at controlling stem elongati on. Later in the crop, drench applications will need to be applied before the presence of visible bud and at higher concentrations than those used in the early dren ches. Applications should be made before the plant begins to elongate ra pidly following transplant. Experiment 5-4 Signif icant cultivar x concentration interactions occurred for phytotoxicity, stem elongation, and number of days to flower (Table 5-4). The interaction was not significant for number of laterals, but the main effects of cultivar and ethephon c oncentration were significant. Phytotoxicity Cultivars res ponded similarly to the application of ethephon at 250 mgL-1 and very little phytotoxicity (marginal chloroisis, marginal necr osis, and/or leaf malformation) was observed (Table 5-5). At 500 mgL-1, all cultivars exhibited some de gree of phytotoxicit y, and AngelMist Dark Lavender and Angelface White Improved we re more severely affected than AngelMist Purple Improved and AngelMist Purple Stripe. At 1000 mgL-1, AngelMist Purple Stripe exhibited the least phytotoxicity sy mptoms relative to the other th ree cultivars. The appearance

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113 of phytotoxicity in all cultivars at the recommende d application concentration is consistent with cultural information published by Proven Winne rs (2006), the supplier of Angelface White Improved. Starman (2001), however, applied et hephon to Blue Pacific and reported an inhibition of stem elongation but did not mention the appearance of any phytotoxicity symptoms. These phytotoxicity symptoms only appeared on th e leaves that were di rectly sprayed with ethephon. By 4 weeks after treatment, damaged leaves were no longer visible under the plant canopy and the leaf morphology of the newly-expanding leaves was normal. Stem elongation Ethephon reduced stem elongation in all cultivar s (Fig. 5-5, Table 5-6) at 4 weeks after treatment. All cultivars ex cept AngelMist Purple Improved exhibited quadratic response curves. The difference between the tallest and sh ortest cultivar at each concentration decreased as the concentration applied increased. AngelMist Dark Lavender and AngelMist Purple Stripe had very similar response curves. Stem elongation was almost identical at 4 weeks after treatment, but most likely would not have continued if data had been taken at a later date due to the presence of flower buds on AngelMist Dark Lavender, an early-flowering cultivar, at the time of data collection. These results are consistent w ith the decrease in stem elonga tion observed in Expt. 5-1. Starman et al. (2004) observed that multiple cultivars of diascia, impatiens, lantana, and nemesia all exhibited decreased stem elongation following the application of ethephon, but in snapdragon, calibrachoa, and petunia, some cultivars responde d and others did not. In this experiment, AngelMist Dark Lavender and AngelMist Purple Stripe ha d a greater response to the ethephon applications than A ngelMist Purple Improved an d Angelface White Improved.

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114 Lateral number The cultivar x concentration interac tion was not significant, but the main effects of cultivar and concentration were significan t (Table 5-7). AngelMist Dark Lavender had a significantly higher number of laterals compared to the other three cultivars. This may be due to the fact that this cultivar flowers earlier than the others, and once angelonia plants begin to develop inflorescences, the axillary meri stems are released from apical inhibition and lateral branching ensues. Since the data was colle cted after AngelMist Dark La vender had initiated flowering, it had started developing secondary laterals in both treated and untreated plants. Ethephon applications at 250 or 500 mgL-1 were not different from the control. Ethephon at 1000 mgL-1 resulted in a higher number of laterals compared to 500 mgL-1. This increase in lateral number is consistent with results from other studies conducted on a number of bedding plants and perennial species (Hayashi et al., 2001; Faust and Lewis, 2005). Flowering The application of ethephon de layed flowering in all cultivars (Fig. 5-6, T able 5-6). AngelMist Dark Lavender and AngelMist Purple Stripe exhibited quadrat ic responses. As the ethephon concentration a pplied increased up to 500 mgL-1, plants exhibited close to a linear increase in time to flower, but between 500 and 1000 mgL-1, the additional amount of delay observed was relatively small. AngelMist Purple Improved and Angelface White Improved exhibited linear responses. AngelMist Dark Lavender was the earliest to flower at all concentrations applied. AngelM ist Purple Improved and Angelface White were the latest to flower at all concentrations appl ied. The untreated controls in bot h cultivars flowered within 2 d of each other, but as the et hephon concentration increased, Angelface White Improved had more of a delay in flowering than A ngelMist Purple Improved. At 1000 mgL-1, it flowered 8 d later.

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115 Delayed flowering in angelonia is consistent with reports of delayed flowering in other ornamental annuals and perennials sprayed with ethephon (Hayashi et al., 2001; Starman et al., 2004). This delay in flowering will inhibit the use of ethepho n as a PGR during crop production. However, it will be beneficial in stock pr oduction, a production situation where it is advantageous to maintain plants in a vegetative state, especially those that are day-neutral. Conclusions Paclobutrazol drenches, dam inozide sprays and ethephon sprays are effective for controlling stem elongation in angelo nia cultivars. Paclobutrazol drenches are activ e at very low concentrations while daminozide and ethephon spra ys require much higher concentrations for efficacy. Response to these PGRs varies with cultivar. For example, Angelina Violet and White was responsive to paclobutrazol, da minozide and ethephon, while AngelMist Dark Lavender was responsive to pacl obutrazol but less sens itive to ethephon and daminozide sprays. Sensitivity to the PGRs was cultivar-specific, bu t was not related to plant vigor. For example, plants of AngelMist Purple Stri pe, one of the most vigorous cultivars in the experiments, drenched with paclobutrazol had a greater percen tage of height control related to untreated plants, while AngelMist Purple Improved, a less vigorous cultivar relative to other angelonia cultivars, had a smaller per centage of height control. The recommended paclobutrazol drench concentration of 5 mgL-1 applied two weeks after transplant (PanAmerican Seed, 2006) was higher than necessary for sufficient control of plant height in Expt. 5-1. However, it was not sufficient for height control in Expt. 5-3, primarily due to it being applied too late in production to control overall plant growth. Drenches should be applied early to mid-way through the cr op, before plants start growing rapidly. Early applications will also help to reduce the amount of lodging present in the ve ry vigorous cultivars, such as AngelMist Purple Stripe and Angelface Blue. Once plants reach visible bud, it is too

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116 late for drench applications to be effective. These experiments were conducted in a warm climate during the summer, and lower concentrations should be effective in cooler temperatures. The recommended rate for daminozide was effective for Angelina Violet and White, the cultivar for which it was recommended (Selecta Fi rst Class, 2008). However, most of the other cultivars were unresponsive or only slightly respon sive to the daminozide sprays. It will need to be applied at higher concentrations or in a tank mix combination with another chemical for sufficient efficacy. Ethephon reduced stem elongation in angel onia cultivars, and the end result was a marketable plant. However, applications result ed in phytotoxicity symptoms in all cultivars. The degree of severity varied depended upon cultiv ar at the low concentrations, but all were severely damaged at 1000 mgL-1. AngelMist Dark Lavender and Angelface White Improved were more sensitive than Ange lMist Purple Improved or Ange lMist Purple Stripe. Ethephon helped increase lateral branching, a positive result in cultivars that have strong apical dominance, but delayed flowering, a negative resu lt in commercial production.

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117 -5 5 15 25 35 45 55 65 05101520 Paclobutrazol concentration (mgL-1)Stem elongation (cm) AM Dk Lav AM Dk Lav Ang. Violet & Wht. Ang. Violet & Wht. AM Purp Imp AM Purp Imp AF Blue AF Blue AM Lav Stripe AM Lav Stripe AF White AF White AM Purp Stripe AM Purp Stripe A -5 5 15 25 35 45 55 65 01250250037505000 Daminozide concentration (mgL-1)Stem elongation (cm) AM Dk Lav AM Dk Lav Ang. Violet & Wht. Ang. Violet & Wht. AM Purp Imp AM Purp Imp AF Blue AF Blue AM Lav Stripe AM Lav Stripe AF White AF White AM Purp Stripe AM Purp Stripe B -5 5 15 25 35 45 55 65 02505007501000 Ethephon concentration (mgL-1)Stem elongation (cm) AM Dk Lav AM Dk Lav Ang. Violet & Wht. Ang. Violet & Wht. AM Purp Imp AM Purp Imp AF Blue AF Blue AM Lav Stripe AM Lav Stripe AF White AF White AM Purp Stripe AM Purp Stripe C Figure 5-1. Effect of plant grow th regulators on stem elongation in Angelonia angustifolia 23 days after treatment. Stem elongation was calculated as the difference between final and initial height. Regression equations (Tab le 5-1) were calculat ed using individual measurements of all plants (n=6). Obse rved points at each c oncentration represent the mean of each cultivar. Plant growth regulators used were A) paclobutrazol B) daminozide C) ethephon.

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118 A B C D E F G H Figure 5-2. Examples of the response of Angelonia angustifolia cultivars to increasing concen trations of paclobutrazol (mgL-1) (Expt. 5-1). The top row is AngelMist Pu rple Stripe, a less sensitive cult ivar. A) 0 B) 5 C) 10 D) 20 mgL-1. The bottom row is Angelina Violet and White, a very sensitiv e cultivar. E) 0 F) 5 G) 10 H) 20 mgL-1.

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119 A B C D E F G H Figure 5-3. Examples of the response of Angelonia angustifolia cultivars to increasing concen trations of daminozide (mgL-1) (Expt. 5-1). The top row is Angelface White, a less responsive cultivar: A) 0 B) 1250 C) 2500 D) 5000 mgL-1. The bottom row is AngelMist Lavender Stripe, a responsiv e cultivar: E) 0 F) 1250 G) 2500 H) 5000 mgL-1.

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120 A B C D E F G H Figure 5-4. Examples of the response of Angelonia angustifolia cultivars to increasing con centrations of ethephon (mgL-1) (Expt. 5-1). The top row is AngelMist Dark Lavender, a less responsive cultivar: A) 0 B) 1250 C) 2500 D) 5000 mgL-1. The bottom row is AngelMist Purple Stripe, a responsive cultivar: E) 0 F) 1250 G) 2500 H) 5000 mgL-1.

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121 0 5 10 15 20 25 30 02004006008001000 Concentration (mgL-1)Stem elongation (cm) AM Dk Lav AM Purp Imp AM Purp Stripe AF White Imp AM Dk Lav AM Purp Imp AM Purp Stripe AF White Imp Figure 5-5. Stem elongation in Angelonia angustifolia cultivars treated wi th ethephon (Expt. 5-4). Data collected at 4W AT. Regression equations (T able 5-6) were calculated using individual measurements of all plants Observed points at each concentration represent the mean of each cultivar (n=9). 0 10 20 30 40 50 60 70 80 90 100 02505007501000 Concentration (mgL-1)Days to first flower AM Dk Lav AM Purp Imp AM Purp Stripe AF White Imp AM Dk Lav AM Purp Imp AM Purp Stripe AF White Imp Figure 5-6. Days to first open flower in Angelonia angustifolia cultivars treated with ethephon (Expt. 5-4). Regression equations (Table 5-6) were calculated using individual measurements of all plants. Observed poi nts at each concentration represent the mean of each cultivar (n=9).

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122Table 5-1. Regression equatio ns for stem elongation for Angelonia angustifolia cultivars in response to PGR concentration (Expt. 5-1). Analysis of variance for treatment, cultivar, and treatment x cultivar were si gnificant at P<0.001. Regression equations were linear (L) or qua dratic (Q) and significant at P 0.05, 0.01, or 0.001. Variable Cultivar Significancez r2 Equation Paclobutrazol AVW L***, Q*** 0.94 y = 47.3 7.10x + 0.246x2 AMDL L***, Q*** 0.96 y = 36.8 3.09x + 0.083x2 AMPI L***, Q*** 0.85 y = 39.6 3.03x + 0.078x2 AMLS L***, Q*** 0.92 y = 44.7 4.74x + 0.140x2 AMPS L***, Q*** 0.94 y = 58.5 5.74x + 0.164x2 AFW L***, Q*** 0.83 y = 54.9 5.31x + 0.158x2 AFB L***, Q*** 0.89 y = 35.7 4.37x + 0.144x2 Daminozide AVW L**, Q* 0.40 y = 47.7 0.008x + 0.000001x2 AMDL L*** 0.53 y = 37.9 0.001x AMPI L*** 0.53 y = 39.5 0.002x AMLS L* 0.45 y = 45.3 0.002x AMPS L***, Q* 0.53 y = 59.3 0.005x + 0.0000005x2 AFW L* 0.24 y = 55.2 0.003x AFB L*** 0.47 y = 38.8 0.003x Ethephon AVW L** 0.34 y = 46.1 0.01x AMDL L*** 0.46 y = 39.6 0.01x AMPI L*** 0.41 y = 37.6 0.01x AMLS L** 0.31 y = 46.3 0.01x AMPS L*** 0.73 y = 59.2 0.03x AFW L* 0.18 y = 54.1 0.01x AFB L***, Q* 0.71 y = 36.1 0.04x + 0.00002x2 z *, **, *** significant at P 0.05, 0.01, and 0.001, respectively.

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123 Table 5-2. Stem elongation (cm) in Angelonia angustifolia four weeks after application of daminozide (Expt. 5-2). The cultivar x con centration interaction was non-significant. Cultivar and concentration were significant at P 0.001 respectively. This table presents the main effects of cultivar and c oncentration. Same letters within each row or column are not significantly different using Tukeys at =0.05. Daminozide concentration (mgL-1) Cultivar 0 1250 2500 5000 Pooled mean (cv) AMDL 46.7 41.6 38.3 35.2 40.5 c AMPS 88.5 76.8 77.0 72.2 78.6 a AFW 52.6 46.3 44.8 38.4 45.5 b Pooled mean (Daminozide) 62.6 a 54.9 b 53.4 b 49.0 c Table 5-3. Stem elongation (cm) in Angelonia angustifolia in response to paclobutrazol applications (Expt. 5-3). Th e early spray was applied 2 weeks after pinch. The late drench was applied at visible bud stage, at 4 weeks after pinch for AngelMist Purple Stripe and at 6.5 weeks for Angelface White. Mean separation was performed using Tukeys at =0.05. Treatmenty Concentration AngelMist Purple Stripe Angelface White Late drench 2 89.3 72.4 Late drench 4 85.8 64.6 Late drench 8 81.2 61.6 Early spray 50 85.8 70.9 Early spray 100 81.9 64.0 Control 83.0 65.7 Pooled means 84.5 a 66.5 b Table 5-4. Analysis of variance for Angelonia angustifolia treated with ethephon (Expt. 5-4). For each treatment, n=12. Phytotoxicity Stem elongation Lateral number Time to flower Cultivar *** *** *** *** Concentration *** *** ** *** Cv. x conc. *** *** NS *** NS,*, **, *** Nonsignificant or significant at P 0.05, 0.01, and 0.001, respectively.

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124 Table 5-5. Mean separation for seve rity of phytotoxicity symptoms in Angelonia angustifolia one week after treatment with ethephon (Expt 5-4). Phytotoxicity was rated on a 0 3 scale, with 0 = no visible symptoms, 1 = slight chlorosis of leaf tips, 2 = moderate chlorosis of leaves and slight necrosis at leaf edges, and 3 = severe chlorosis of leaves, moderate to severe leaf necrosis, and leaf distortion. Mean separation was performed using Tukeys at =0.05, and HSD =0.05 = 0.6. Cultivar Ethephon (mgL-1) AngelMist Dark Lavender AngelMist Purple Improved AngelMist Purple Stripe Angelface White 0 0.0 c 0.0 c 0.0 c 0.0 c 250 0.1 c 0.1 c 0.3 c 0.2 c 500 2.7 a 1.8 b 1.4 b 2.9 a 1000 2.9 a 2.7 a 2.0 b 2.9 a Table 5-6. Regression equations for Angelonia angustifolia cultivars treated with ethephon (Expt. 5-4). Variable Cultivar Significancez r2 Equation Stem elongation (cm) AMDL L***, Q*** 0.78 y = 25.8 0.03x + 0.00002x2 AMPI L** 0.29 y = 12.6 0.005x AMPS L***, Q** 0.78 y = 24.8 0.03x + 0.00002x2 AFWI L***, Q** 0.59 y = 11.0 0.01x + 0.00001x2 Days to flower AMDL L***, Q*** 0.80 y = 33.4 + 0.06x 0.00004x2 AMPI L*** 0.36 y = 66.8 + 0.0124x AMPS L***, Q** 0.68 y = 54.7 + 0.03x 0.00002x2 AFWI L*** 0.67 y = 65.7 + 0.02x z NS,*, **, *** Non-significant or significant at P 0.05, 0.01, and 0.001, respectively. Table 5-7. Mean separation for number of laterals in Angelonia angustifolia 4 weeks after ethephon treatment (Expt. 5-4). Mean separation was performed using Tukeys at =0.05. HSD values for the main effects of both cu ltivar and concentration were 1.7. Same letters within each row or column are not significantly different. Ethephon concentration (mgL-1) Cultivar 0 250 500 1000 Pooled means (cv) AMDL 19.8 20.7 20.7 22.7 20.9 a AMPI 3.8 4.0 5.1 7.0 4.8 b AMPS 6.3 4.2 4.6 6.3 5.4 b AFWI 3.4 3.9 5.8 6.3 4.9 b Pooled means (ethephon) 8.3 b 8.2 b 9.1 ab 10.8 a

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125 CHAPTER 6 CONCLUSIONS The increase in popularity of Angelonia angustifolia has led to the developm ent and release of cultivars with novel grow th habits and flower color, and companies are including these new releases within their available product line, ofte n referred to as a series. The availability of new cultivars into an existing product line does not guarantee that all of them will have similar growth habits or production requirements, which can be a problem for growers. In production, it is optimal to select cultivars with similar pr oduction requirements, such as optimal fertilizer concentration and like sensitivities to PGRs. Al so, time to flower will determine the length of the production schedule, and grower s would prefer cultivars with shorter production times so that more turns can be accomplished in a greenhouse during a season. Thirty-one Angelonia angustifolia cultivars we re observed for differences in growth habit and flowering traits. Differences of more th an 3 weeks were observed between cultivars for number of days to first flower and number of days to marketabilit y. Plant height at marketability ranged from 14 to 64 cm, and plant width ranged from 28 to 54 cm. Cultivars with naturally tall or wide growth habits will be difficult to grow in production without the us e of PGRs to inhibit stem elongation. Another issue in production is the susceptibility of certain cultivars to powdery mildew. Plants were grown in a naturally ventilated greenhouse and not treated with any fungicides for the duration of the experiment. In some cultivars, no powdery mildew was detected, while in others, it severe ly affected plant quality. The range of susceptibility observed may indicate that it could be a trait that breede rs can select for in their breeding programs. Angelonia is a warm season crop produced during the early spring in southern states and in late spring for the northern states. While angelonia are not photoperiodic, they respond to changes in temperature and light quantity associated with seasonality. From late winter to early

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126 summer, cultivars flowered 17 to 29 d earlier, indi cating that temperature and light most likely play a major role in hastening flower development. In cooler climates, the cost of supplemental heating and lighting may be cost -effective if the crop time can be shortened significantly. Angelonia cultivars responded similarly to incr easing applications of fertilizer and can be treated uniformly in a greenhouse. This is beneficial to large-scal e growers who may have multiple annual species or cultivars in a gree nhouse section. They respond well to a complete fertilizer with N at 100 mgL-1, which is similar to many other warm season annuals (Kang and van Iersal, 2002; Kent and Reed, 1996). Although this is less th an recommended for other crops, such as petunia (James and van Iersal, 2002), cult ivars did not show any sensitivity to increased fertilizer concentrations other than the appearan ce of darker green leaves and a slight reduction in growth. Angelonia cultivars, however, are cultivar-specific in response to PGRs. Daminozide is effective at 5000 mgL-1 in Angelina Violet and White a nd Angelface Blue but other cultivars are relatively insensitive. Pacl obutrazol is ineffective as a spray, but is highly effective as a drench if applied early in the crop. Ethephon is effective at inhibiti ng stem elongation, but the potential for phytotoxicity is greater in certain cultivars. None of the cultivars show permanent damage from ethephon, but its ability to delay flowering can be an issue. Angelonia cultivars show a wide range of heat tolerance with respect to flowering during the summer months. The earliest plant date, 4 Apr. 2007, provided the best display of color when all of the cultivars reached their peak flowering ability. In the latest plant date, none of the cultivars were able to get established adequately and provide a similar display of color. All of the cultivars were negatively affected by the high temperatures, but AngelMist Purple Stripe and Angelface White were less affected.

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127 In greenhouse production, AngelMist Dark Lave nder is an ideal cultiv ar to grow due to its early flowering and compact plant habit, which requires minimal growth regulation to keep its size under control. However, it is very sens itive to powdery mildew and will need to be monitored during the spring growing season. In th e landscape, it will flower in late spring and early summer, but not stay in flower throughout the summer. This is an example of a greenhouse-friendly cultivar but not a landscape-friendly cultivar. On the other side of the spectrum is AngelMist Purple Stripe. It is a me dium to late flowering cultivar, and it is one of the most vigorous angelonia cultivars available. It also has very sticky, oily, pubescent leaves and stems, which makes the application of PGRS which are necessary to keep it from becoming overgrown, difficult. However, in the landscape, it has excellent heat tolerance and will provide a consistent display of colo r throughout the summer season. Our study complements the studies published by Miller and Armitage (2002) and Miller et al. (2001) on angeloni a production. However, many gaps s till exist, including the specific effects of temperature and light on flowering. What is the rela tive impact of temperature and flowering on flower initiation and development, and can this be quantified such that a grower can know exactly how to hasten or delay marketability of the crop? In certai n cultivars, such as AngelMist Purple Stripe and Angelface White, it appears that certain traits may be linked; for example, these were the only cul tivars with sticky pubescent leaves and also they were the only two to continuously flower throughout the summer, which may indicate heat tolerance relative to other cultivars with respect to floral development. Also, Angelface White was the latest cultivar to flower and AngelMist Purple Stripe was one of the late r-flowering cultivars, possibly indicating that these two may have a higher temperature optimum for flower development and continued flowering.

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128 Angelonia is a crop with breedi ng potential. The range of va riability present within this species indicates that many of th ese traits can be selected and improved upon such that a cultivar that is easily grown and produced and will provide good landscape color during the season. In addition, A. angustifolia is the only species known to curren tly be in commercial production and related species may hold cultivation potential or have desirable traits which could be incorporated into a breeding program.

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138 Williams, A.L. 2004. Foliar symptomology and tissu e concentrations of five nutritionally deficient floriculture crops. Masters Thesis, N.C. State Univ. Willits, D.H. and D.A. Bailey. 2000. The effect of night temperature on chrysanthemum flowering: heat-tolerant ve rsus heat-sensitive cultiv ars. Scientia Hort. 83:325-330.

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139 BIOGRAPHICAL SKETCH Jennifer Kay Boldt was born in Minnesota and rais ed in Florida. She and her twin sister Jessica em braced horticulture at a young age and grew up helping their parents in the familyowned nursery and garden center. She earne d a Bachelor of Science in environmental horticulture and a Bachelor of Arts in business ad ministration from the University of Florida in December 2005. As an undergraduate, Jennifer completed two horticulture-related internships: a six-month production internship wi th Van Wingerden International in Fletcher, North Carolina, and a three-month trial garden internship with Ball Horticultural Company in West Chicago, Illinois. As a graduate student, she was a resear ch assistant and coordina ted the University of Floridas on-campus floriculture tr ial gardens. Jennifers future career plans lead her back to Minnesota, where she will begin a PhD program in Fall 2008.


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