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Stock Plant Management of Tropical Perennials

HIDE
 Title Page
 Dedication
 Acknowledgement
 Table of Contents
 List of Tables
 List of Figures
 Abstract
 Literature review
 Stock plant nutrition
 Lateral branching agents
 Rooting hormones
 Research summary
 References
 Biographical sketch
 

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STOCK PLANT MANAGEMENT OF TROPICAL PERENNIALS By CHRISTOPHER BRIAN CERVENY 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 2006

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Copyright 2006 by Christopher Brian Cerveny

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This document is dedicated with expresse d appreciation to the “Research Machine” Cricket, Dawn, Jude, Leah, and Neil.

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iv ACKNOWLEDGMENTS I thank my graduate committee, Drs. Jame s E. Barrett, Brian E. Myers, and James L. Gibson, for their guidance and support throug hout the degree process. I thank Carolyn Bartuska and Dr. Grady L. Miller for stat istical assistance. For numerous hours of assistance during experiment prep arations and collection of da ta, I wish to thank Jeffrey Anderson, Shannon Crowley, Neil Greishaw, J ude Groninger, Alina Lovelace, Leah McCue, Meghan Pressley, Dawn Taylor, Cars on Winn, and Mariah Williams. Additional appreciation is expressed to Bob Weidman for assistan ce with growth regulator applications and for providing st udent labor at even the shorte st of notice. I thank the Fred C. Gloeckner Foundation for grant support and Hatchett Creek Farms for donation of plant materials. For chemical dona tions throughout the experiment process appreciation is given to Fine Agrochemicals, Monterey Ga rden Supply, and Dip n’ Grow Inc. Lastly, I wish to thank my parents for expressing pride in my accomplishments. This was a source of motivational support that is without comparison.

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v TABLE OF CONTENTS page ACKNOWLEDGMENTS.................................................................................................iv LIST OF TABLES...........................................................................................................viii LIST OF FIGURES..........................................................................................................xii ABSTRACT.....................................................................................................................xi ii CHAPTER 1 LITERATURE REVIEW.............................................................................................1 Introduction to the Floriculture Industry......................................................................1 Asexual Propagation of Cuttings..................................................................................3 Physiological Age of Cuttings...............................................................................3 Growing Environment...........................................................................................4 Mineral Nutrition...................................................................................................5 Growth Regulator Treatments...............................................................................6 Rooting Hormone Treatments...............................................................................7 Conclusion.............................................................................................................8 Selected Tropical Species.............................................................................................9 Allamanda schottii .................................................................................................9 Bougainvillea glabra ...........................................................................................10 Mandevilla splendens ..........................................................................................11 Nerium oleander ..................................................................................................12 Tecoma stans .......................................................................................................13 Objectives...................................................................................................................13 2 STOCK PLANT NUTRITION...................................................................................14 Introduction.................................................................................................................14 Materials and Methods...............................................................................................15 Experiment 1.......................................................................................................15 Experiment 2.......................................................................................................19 Results........................................................................................................................ .19 Experiment 1.......................................................................................................19 Stock Plant Parameters........................................................................................19 Cutting quantity............................................................................................19

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vi Cutting length, stem diameter, and leaf area................................................20 Rooting Parameters.............................................................................................21 Percent survival, visual rooting quality, and quantity of primary and lateral roots...............................................................................................21 Cutting dry weight and root : shoot ratio.....................................................24 Stock plant leachate......................................................................................26 Foliar analysis..............................................................................................27 Experiment 2.......................................................................................................28 Stock plant parameters........................................................................................28 Cutting quantity............................................................................................28 Cutting length, stem diameter, and leaf area................................................29 Rooting parameters..............................................................................................31 Percent survival, visual rooting quality, and quantity of primary and lateral roots...............................................................................................31 Cutting dry weight and root : shoot ratio.....................................................33 Stock plant leachate......................................................................................34 Foliar analysis..............................................................................................34 Discussion...................................................................................................................36 Conclusion..................................................................................................................41 3 LATERAL BRANCHING AGENTS.........................................................................55 Introduction.................................................................................................................55 Materials and Methods...............................................................................................56 Experiment 1.......................................................................................................56 Experiment 2.......................................................................................................57 Harvesting procedure...................................................................................57 Cutting evaluations.......................................................................................58 Results........................................................................................................................ .58 Experiment 1.......................................................................................................58 Harvest yield, cutting length, and stem diameter.........................................58 Percent survival and vi sual rooting quality..................................................62 Cutting total dry weight and root : shoot ratio.............................................63 Experiment 2.......................................................................................................65 Harvest yield, cutting length, and stem diameter.........................................65 Percent survival and vi sual rooting quality..................................................69 Cutting total dry weight and root : shoot ratio.............................................71 Discussion...................................................................................................................74 Conclusion..................................................................................................................79 4 ROOTING HORMONES...........................................................................................81 Introduction.................................................................................................................81 Materials and Methods...............................................................................................82 Experiment 1.......................................................................................................82 Harvesting procedure...................................................................................83 Cutting evaluations.......................................................................................83

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vii Experiment 2.......................................................................................................84 Results........................................................................................................................ .85 Allamanda schottii ...............................................................................................85 Experiment 1................................................................................................85 Experiment 2................................................................................................91 Discussion....................................................................................................96 Conclusion....................................................................................................98 Bougainvillea glabra ‘California Gold’..............................................................98 Experiment 1................................................................................................98 Experiment 2..............................................................................................103 Discussion..................................................................................................105 Conclusion..................................................................................................107 Bougainvillea glabra ‘Helen Johnson’..............................................................107 Experiment 1..............................................................................................107 Experiment 2..............................................................................................112 Discussion..................................................................................................118 Conclusion..................................................................................................119 Mandevilla splendens ‘White’...........................................................................120 Experiment 1..............................................................................................120 Experiment 2..............................................................................................121 Discussion..................................................................................................123 Conclusion..................................................................................................124 Nerium oleander ‘Dwarf Salmon’.....................................................................125 Experiment 1..............................................................................................125 Experiment 2..............................................................................................129 Discussion..................................................................................................134 Conclusion..................................................................................................136 5 RESEARCH SUMMARY........................................................................................138 LIST OF REFERENCES.................................................................................................141 BIOGRAPHICAL SKETCH...........................................................................................145

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viii LIST OF TABLES Table page 2-1 Temperature and relative humidity da ta (maximum, minimum, and mean) as recorded during simulate d and actual shipping........................................................42 2-2 Influence of harvest date, cultivar, and nitrogen (N) concentration on quantity of sub-terminal bougainvillea stem cuttings …............................................................42 2-3 Significance levels for effects of harvest date, cultivar, and nitrogen (N) concentration on length and leaf area of cuttings.....................................................43 2-4 Harvest date (H) by nitrogen (N) con centration interacti on for length of subterminal bougainvillea stem cuttings........................................................................43 2-5 Significance levels for effects of ha rvest date, evaluation date, cultivar, and nitrogen concentration..............................................................................................44 2-6 Harvest date (H) by nitrogen (N) concen tration interaction for percent survival of sub-terminal bougainvillea...................................................................................44 2-7 Harvest date (H) by cultivar by N con centration interaction for visual rooting quality (1=poor; 5=best)...........................................................................................45 2-8 Significance levels for effects of harvest date, cultivar, and nitrogen concentration............................................................................................................45 2-9 Harvest date by cultivar by N concentr ation interaction for total cutting dry weight (shoot weight + root weight)........................................................................45 2-10 Significance levels for effects of harvest date, cultivar, and nitrogen (N) concentration on foliar concentration.......................................................................46 2-11 Harvest date (H) by cul tivar interaction for foliar nutrient concentration of macronutrients..........................................................................................................46 2-12 Harvest date (H) by nitrogen (N) con centration interaction for foliar nutrient concentrations of macronutrients.............................................................................47 2-13 Cultivar by nitrogen (N) concentration interaction for foliar nutrient concentration of macronutrients...............................................................................47

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ix 2-14 Influence of harvest date, cultivar, and nitrogen (N) concentration on quantity of sub-terminal bougainvillea stem cuttings….............................................................48 2-15 Significance levels for effects of harvest date, cultivar, and nitrogen (N) concentration............................................................................................................48 2-16 Cultivar by nitrogen (N) concentration interaction for leaf area of sub-terminal bougainvillea stem cuttings harvested during the cool season (Expt. 2)..................49 2-17 Significance levels for effects of ha rvest date, evaluation date, cultivar, and nitrogen concentration..............................................................................................49 2-18 Harvest date (H) by cultivar by nitrogen (N) concentration interaction for visual rooting quality..........................................................................................................50 2-19 Influence of harvest date (H), cu ltivar, and nitrogen (N) concentration on quantity of primary roots..........................................................................................50 2-20 Significance levels for effects of harvest date, cultivar, and nitrogen (N) concentration on foliar concen tration of macronutrients.........................................51 2-21 Harvest date (H) by cultivar by nitrogen (N) concentration interaction for foliar nutrient concentration...............................................................................................51 3-1 Significance levels for effects of harv est date, cultivar, and branching treatment on quantity, length, and stem diameter…................................................................59 3-2 Cultivar by treatment interaction for qua ntity and length of 2-node stem cuttings.59 3-3 Significance levels for effects of harv est date, cultivar, and lateral branching treatment on percent survival visual rooting quality...............................................63 3-4 Harvest date by cultivar by branching treatment inte raction for rooting quality (1=poor; 5=best) and percent survival.....................................................................64 3-5 Significance levels for effects of harv est date, cultivar, and branching treatment on quantity, length, and stem diameter of cuttings harvested from stock plants of oleander and tecoma during th e cool season (Expt. 1).............................................65 3-6 Influence of lateral branching agents on number and lengt h of cuttings and subsequent rooting performance..............................................................................66 3-7 Influence of lateral branching agen ts on quantity and le ngth of cuttings and subsequent dry weights............................................................................................67 3-8 Influence of lateral branching agen ts on quantity and le ngth, and subsequent rooting......................................................................................................................70

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x 3-9 Influence of lateral branching agents on number and lengt h of cuttings, and subsequent rooting performance..............................................................................71 3-10 Significance levels for effects of harv est date, cultivar, and lateral branching treatment on percent survival visual rooting quality…...........................................80 4-3 Harvest date (H) by treatment interacti on for visual rooting quality (1-5; 1=poor, 5=best) of allamanda cuttings planted during the warm season (Expt. 1)...............86 4-4 Harvest date by treatment interaction for number of primary roots of stem cuttings of allamanda planted during the warm season (Expt 1).............................88 4-5 Harvest date (H) by treatment intera ction for number of lateral roots of allamanda cuttings planted duri ng the warm season (Expt. 1).................................88 4-6 Harvest date by treatment interaction fo r total cutting dry weight of allamanda cuttings planted during the warm season (Expt. 1)..................................................91 4-7 Harvest date by treatment interaction for the cutting root : shoot ratio of allamanda cuttings planted duri ng the warm season (Expt. 1).................................91 4-9 Harvest date by treatment interaction fo r number of primary roots of allamanda cuttings planted during the cool season (Expt. 2)....................................................94 4-2 Influence of rooting hormone applic ation on percent survival of allamanda cuttings evaluated.....................................................................................................99 4-8 Influence of rooting hormone applic ation on the visual rooting quality of allamanda cuttings evaluated.................................................................................100 4-10 Influence of rooting hormone on cutting root : shoot ratio of allamanda cuttings evaluated.................................................................................................................101 4-11 Harvest date (H) by treatment intera ction for bougainvill ea ‘California Gold’ cuttings planted during the warm season (Expt. 1)................................................104 4-12 Harvest date (H) by treatment interacti on for root : shoot ra tio (root weight + shoot weight) of ‘Helen Johnson’ bougainvillea....................................................110 4-13 Harvest date (H) by treatment interacti on for visual rooting quality (1-5; 1=poor, 5=best) of bougainvillea.........................................................................................116 4-14 Harvest date (H) by treatment intera ction for number of primary roots of bougainvillea ‘Helen Johnson’ cuttings pl anted during the cool season (Expt. 2).116 4-15 Harvest date (H) by treatment intera ction for number of lateral roots of bougainvillea ‘Helen Johnson’ cuttings pl anted during the cool season (Expt. 2).117

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xi 4-16 Harvest date by treatment interaction for visual rooting quality (1-5; 1=poor, 5=best) of mandevilla cuttings plante d during the cool season (Expt. 2)..............122 4-17 Harvest date (H) by treatment interacti on for visual rooting quality (1-5; 1=poor, 5=best) of oleander cuttings plante d during the warm season (Expt. 1)................126 4-18 Harvest date (H) by treatment interacti on for number of lateral roots of oleander cuttings planted during the warm season (Expt. 1)................................................128 4-19 Harvest date (H) by treatment interacti on for total cutting dry weight of oleander cuttings planted during the warm season (Expt. 1)................................................129 4-20 Harvest date (H) by treatment interacti on for visual rooting quality (1-5; 1=poor, 5=best) of oleander cuttings plante d during the cool season (Expt. 2)..................131 4-21 Harvest date (H) by treatment intera ction for number of primary roots of oleander cuttings planted duri ng the cool season (Expt. 2)....................................132 4-22 Harvest date (H) by treatment interacti on for number of lateral roots of oleander cuttings planted during the cool season (Expt. 2)..................................................133 4-23 Harvest date (H) by treatment interacti on for root : shoot ra tio (shoot dry weight / root dry weight) of oleander cuttings pl anted during the cool season (Expt. 2)..134 4-1 Temperature and relative humidity data as recorded during simulated and actual shipping..................................................................................................................137

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xii LIST OF FIGURES Figure page 2-1 Nitrogen (N) concentration x sampling da te interaction for leachate pH collected from bougainvillea stock plants fertilized with........................................................52 2-2 Main effects for leachate electrical conductivity (EC) over time, as recorded from bougainvillea stock plants duri ng the warm season (Expt. 1).........................52 2-3 Nitrogen (N) concentration x sampling date interaction for nitrate nitrogen (NO3-N) values recorded from stock plant leachate................................................53 2-4 Nitrogen (N) concentration by sampli ng date interaction for leachate pH collected from bougainvillea stock plants................................................................53 2-5 Main effects for leachate electrical conductivity (EC) over time as recorded from bougainvillea stock plants during the..............................................................54 2-6 Nitrogen (N) concentration by sampling date interaction for leachate nitrate nitrogen (NO3-N) values recorded from stock plants...............................................54

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xiii 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 STOCK PLANT MANAGEMENT OF TROPICAL PERENNIALS By Christopher Brian Cerveny August 2006 Chair: James L. Gibson Major Department: Environmental Horticulture A series of greenhouse-based propagation experiments were conducted during the 2005 summer (warm season) and 2005-2006 fall/w inter (cool season) to investigate the influence of nutrition, chemical branching ag ents, or rooting hormone treatments on the stock plant yield of tropical perennials and subsequent rooting performance of cuttings. In the first set of experiments stock plants of Bougainvillea glabra (Choisy.) 'Raspberry Ice' and 'Purple Small Leaf' were fertilized with three concentrati ons of nitrogen (N) at 100, 200, and 300 mgL-1 to determine optimum N fertig ation standards for maximizing yield of high quality stem cutti ngs that root the best. Of the concentrations studied, the range of 200 to 300 mgL-1 was shown to effectively increase cutting yield and improve subsequent rooting performance. Concen trations should be provided at 300 mgL-1 during the active growth periods associat ed with summer, and reduced to 200 mgL-1 as daylength becomes shorter, temperatures de crease, and light quant ity is reduced during fall/winter months. In a second set of experiments, stock plants of Tecoma stans (L.) Juss. ‘Esperanza’ and Nerium oleander (L.) ‘Dwarf Salmon’ were sprayed with

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xiv commercially available lateral branching agents to identify appropriate treatments for increasing yield and enhanc ing rooting performance. Results indicated that concentrations applied may have been excessi ve, as phytotoxic symptoms were shown to damage or distort cutting tissue and limit r ooting. Fresco, although highly phytotoxic to both species, tended to increase cutting yi eld in oleander. FAL-457 was shown to increase yield; however rooting was hindered. Florel was shown to increase yield with similar rooting to the control; unfortunately cutting stunting and tipburn occurred at the concentrations applied. Furt her investigation with these species should be conducted at lower concentrations or with combination la teral branching agent mi xtures. In a third series of experiments several species of tr opical perennials were treated with rooting hormones to measure the effect of synthetic a uxin on rooting. Treatments consisted of an untreated control, a water-sol uble rooting hormone indole-3butyric acid, potassium salt (KIBA) applied at concen trations of 1500 to 6000 mgL-1, or the standard alcohol-based Dip N’ Grow at 1500 mgL-1. Results indicated KIBA was an effective formulation for propagation of alcohol-sensitive species such as oleander, an d was comparable to Dip N’ Grow. Although concentrations varied with species invest igated, a recommended KIBA range of 3000 to 6000 mgL-1 was shown to be most effective for increasing cutting rooting performance.

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1 CHAPTER 1 LITERATURE REVIEW Introduction to the Floriculture Industry The U.S. Floriculture Industry has changed in several ways over the past 50 years. From garden plants to house plants and cu t flowers, florists no longer are the primary distributor for horticultural pr oducts. Presently, floriculture crops are marketed through a wide variety of sales outlets, from retail gr eenhouses and garden centers, to supermarkets, large chain stores, and seasonal road-s ide stands (Ball, 1998; Hamrick, 2003). Today’s ornamental plant industry offers a large number of plant species and cultivars for sale including: traditional bedding plants, trees a nd shrubs, vegetables, ground covers, and foliage crops. However, fr om a research and development point of view, perennials are the most popular crop in the North Amer ican market. This broad classification of herbaceous plants has pr oduced an abundant amount of research and production information over the past 15 year s (Nau, 1999). In 2004 production totals of perennials were reported at $687 million, up 8% from the previous year (NASS, 2004), and are the fastest-growing se gment of floriculture crops. As herbaceous perennial production becomes mo re efficient, plants are able to be mass produced with less capital input. Thus, perennials ar e becoming available at the low priced, “big box” stores, which mercha ndise nearly 75% of horticultural products (Wolnik, 1994). Faced with a highly compe titive market and narrow profit margins, the U.S. Bedding Plant Industry has shifted from the traditional packaging of multi-celled, inexpensive units with many plants per containe r, to larger pots th at fetch a higher price

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2 per individual plant, some of which include: 4.5-inch round -pots, trade-gallon nursery containers, and various specialty container gardens including color bowls, window boxes, and patio planters (Ball, 1998; Hamrick, 2003). This shift has brought an increased interest in using temperate climate herbaceous perennials for winter landscapes in the South; unfortunately the excessively high temperatures of summer can negatively im pact those plants that perform well under USDA Hardiness Zones 4 through 7. Hence, tropical perennials are growing in popularity. These versatile plants can be used as hardy perenn ials in southern climates and as summer annuals in the North. Annuals have often been described as us eful enhancements to landscape designs through foliar textures, flower colors, and plan ting patterns that can be changed each year (Arent and Voigt, 1994). Perennial plant viewpoints differ amongst various regions of the country, and what may be considered a perennial in one lo cation may likely be considered an annual in another (Nau, 1999). Regardless of these differences in opinion or performance of perennials, there has been an emerging trend for gardeners to purchase and trial new and exciting plants. Because the popularity of gardening with tr opical plants has grown in the U.S., there has been a considerable increase in production and sa les of tropical annuals and perennials over the past several years (Bowden, 2004). However, little cultural information is known on how to reproduce thes e plants effectively, so growers tend to use some degree of trial and error (G. Griffith, Hatchett Creek Farms, personal communication). This literatu re review will focus on how tr opical plants are asexually

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3 propagated by stem cuttings, with an emphasi s on select tropical species, and how the stock plants are managed. Asexual Propagation of Cuttings Vegetative or asexual propagation has become an established system for reproducing plants which exhibit desirable characteristics. The common methods for asexual propagation include using roots, stem s, or leaves of stock plants for grafting, tissue culture, division, or cutting propaga tion (Davidson et al., 2000). In general, propagation by stem cuttings has numerous a dvantages. Many plants can be grown in high density trays from a limited amount of st ock plants. As compared to other asexual means of reproduction, it is typically less e xpensive, quicker, relatively simple, and does not require mastery of di fficult techniques such as grafting, budding, or micropropagation (Hartmann et al., 2002). Stem cutting production involves removing a shoot tip from a mother plant and planting it in a growing substrate to r oot. This method allows for retention of foliar, flowering, and growth habit characteristics that may not be carried over via seed production. Successful propagation of landscape plants by rooting vegetative stem cuttings depends on several factors, including: phys iological state of stem cuttings, the propagation environment, fertility management and growth regulator treatments, whether applied to the stock plant prior to harvesti ng or exogenously applied rooting hormones to stem cuttings (Atzmon et al., 1997; Hartmann et al., 2002) Physiological Age of Cuttings The physiological age of plant material harvested for propagation has been documented as an important factor in some tropical species. Terminal stem or tip cuttings include: the stem apex or shoot tip, developing immature leav es, and one or more

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4 mature leaves. These cuttings generally pr oduce a more efficient crop, by effectively rooting in the quickest time (D ole and Wilkins, 1999). Older stem tissue is frequently slower to root than young tissue, but gene rally cuttings with thick stems and short internodes will produce the best plants. Th ere is little information published on how to root tropical cuttings or at which physiological age the tissue should be. However, some species such as bougainvillea ( Bougainvillea glabra Choisy) root better from semihardwood to hardwood cuttings depending on temperature (Schoellhorn and Alvarez, 2002), location (Auld, 1987), and time of year (Chakraverty, 1970). Growing Environment Propagation success also depends on the level to which a suitable growing environment is offered. The propagation substr ate temperature, ambient air temperature, and relative humidity levels should be car efully monitored during propagation, whether the plant material is herbace ous, softwood, semi-hardwood, or hardwood tissue. Tropical plants such as bougainvillea benefit from bottom soil temperatures of 30 C (86 F) (Singh et al., 1976). If bottom heat is provided, so il thermometers or remo te recording sensors should be installed within th e root zone area and checked frequently. Excessively high temperatures in substrate, even for a short ti me, are likely to result in damage to basal tissue (Hartmann et al., 2002). In addition to temperature, it is important to maintain high humidity levels while rooting vegetative stem cuttings in order to reduce water loss due to transpiration. Intermittent mist should be used to mainta in high humidity levels, but unfortunately prolonged periods of mist can leach mineral nut rients (Hartmann et al., 2002). In most cases, cuttings benefit from eith er a top dressing of slow rele ase fertilizer or a low level application of liquid fertilizer after roots begin to emerge fr om the cutting base. Because

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5 some tropical plants often require extensive ro oting time in propagation, the risk of air and waterborne pathogens also increases (Howard, 1994). Mineral Nutrition Mineral nutrition is one of many factors influencing fo rmation of adventitious roots in cuttings. Rooting stages can be generalized into two categories: 1) root initiation; and 2) root growth and development (Blazic h, 1988b). Therefore, when considering the influence various mineral nutrients have on adventitious root formation, one should consider the function of these elemen ts during each stage of development. Root initiation involves dedifferentiation of specific cells, leading to the formation of root meristems and is dependent on th e presence of auxin, wh ether endogenous or artificially applied. Most studies have not given a clear understand ing of the importance of specific nutrients in the initiation of adve ntitious roots; however one could argue that any nutrient essential for root initiation is also necessary for plant growth and development. For example, nitrogen (N) content is crucial in cuttings as it is important for nucleic acid and protein synthesis in plan t tissue (Hartmann et al., 2002). Nitrogen concentration also affects cutting yield of stock plants and rooting performance of cuttings. When considering the role N has in metabolic processes, one could strongly argue its importance in root initiation (Blazic h, 1988b). While excessive N has been shown to negatively affect cutting propa gation (McAvoy, 1995), heavy fertilization in some tropical plants such as bougainvillea, ha s been observed to i nhibit flowering as it promotes vegetative shoot growth (Schoellhorn and Alvarez, 2002). While the role of specific elements in r oot initiation remains unclear, the nutrition of stock plants seems to have a more notew orthy effect. Although th is influence on stock plants of tropical perennials is not we ll documented, nutrient le vel concentrations

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6 recommended for other floriculture crops ha ve been studied. Dreuge et al. (2000) reported a positive correlation with the number and length of roots of propagules when stock plants of chrysanthemum ( Dendranthema x grandiflorum Kitam.) were treated with increased N concentration rates from 60 to 400 mg.L-1. In another experiment, Roeber and Reuther (1982) showed a negative correl ation in cutting production with increasing rates of N fertilization, when concentrations of 56, 112, or 168 mg.L-1 were applied to stock plants of chrysanthemum grown in nutri ent solution. In a study where stock plants of geranium ( Pelargonium x hortorum L. H. Bailey) were fertilized at N concentrations of 50, 100, 200, and 400 mg.L-1, respectively, higher concentrations of N produced similar results in number of vegetative cu ttings generated, but with N at 50 mg.L-1, lower numbers of vegetative cuttings develope d (Ganmore-Neuman, 1990; Ganmore-Neuman, 1992). Although optimum nutrient standards for tropi cal stock plants remain undefined, as with stock plants from other floriculture cr ops, it is essential that the propagules come from a nutritionally healthy source (Hartma nn et al., 2002). Nutrition studies on stock plants of tropical perennials should be c onducted to determine optim um concentrations of N fertilization necessary to produce the greate st number of high quality cuttings with subsequently greater propagation su ccess and cuttings establishment. Growth Regulator Treatments Some naturally occurring pl ant hormones such as ethyl ene and cytokinin can be used as growth regulator treatments to pr omote lateral branching. The effects plant growth regulators (PGRs) ethephon [(2-chloroethyl) phosphoni c acid] (Florel) (Monterey Lawn and Garden Products, Inc., Fresno, Ca lif.) and benzyladenine (BA) (cytokinin) have on stock plant cutting quantity has been investigated and has been proven to be

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7 beneficial for promoting axillary shoot devel opment in some floricultural crops such as: chrysanthemum; geranium; poinsettia ( Euphorbia pulcharrima Willd. ex Klotsch); and vinca vine ( Vinca major L.) (Konjoian, 1994; Witaszek and Mynett, 1989). In foliar applications of ethephon to stock plants of Scaevola ( Scaevola aemula R. Br ) cutting length was shown to be longer with ethephon foliar sprays of 250 to 1000 mg.L-1 than the untreated control (Gibson and Whipker, 2004). There is some evidence that cytokinin applications assist in adve ntitious root formation. Improved shoot development and rooting is achieved by its applic ation to leaves and buds of the stock plant, rather than to the basal tip of stem cuttings (Davis et al ., 1988). However in most cases cytokinin application to plants will i nhibit adventitious root formation as it increases lateral branching (Mynett, 1985). Rooting Hormone Treatments Countless studies have reported on the s timulatory influence of auxin on the propagation of cuttings of diffi cult-to-root species. Treating cuttings with auxin increases rooting percentage, hastens root formation, and increases uniformity of rooting (Davis et al., 1988). Indole acetic acid (IAA) is the natu rally occurring auxin found in plants and has been documented in nearly every aspect of plant growth and development. Some of the processes regulated by IAA, as they pertain to tropical pl ant propagation include: induction of cell division, stem elongation, ap ical dominance, induc tion of rooting, and vascular tissue differentia tion (Srivastava, 2002). Synt hetic forms of auxin are commercially available in the form of indole butyric acid (IBA) and napthaleneacetic acid (NAA). Preference for these sythentic compounds compared to IAA, is illustrated by the large number of rooting products containing IBA, NAA, or in combination (Blazich, 1988a).

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8 Auxin containing rooting hormones can be a pplied to the base of cuttings as talcbased powder or dipped in a liquid solution containing 50% ethanol or isopropyl alcohol and 50% water. Cuttings are dipped for a pe riod of time from a few seconds to 12 hours (Dole and Wilkins, 1999). Liquid treatments us ually provide the most consistent results because application is more uniform than with powder. However, powder dips are sometimes preferred since it decreases the chance for water-borne disease transmittance. There are also potassium salt (K+) formul ations that enable IBA (KIBA) and NAA (KNAA) to be dissolved in wa ter (Hartmann et al., 2002). Sometimes mixing auxin with other car riers such as polyethylene glycol (antifreeze), propylene glycol, or windshield wa sher fluid may facilitate auxin application on difficult-to-root species which require high levels of hormone to induce rooting, but with sensitivity to alcohol. However these methods often involve heating the solutions to 72 C (161.6 F), which facilitates dissolving th e auxin into polyethyl ene glycol causing the materials to not disassociate after cool ing (Hartmann et al., 2002). Often nurseries use KIBA with vegetative cuttings during active growth stages and IBA with ethanol during dormant periods. Research has suggested some tropi cal species such as oleander ( Nerium oleander L.) and bougainvillea benefit from applications of 3,000 mg.L-1 IBA and 4,000 to 16,000 mg.L-1 IBA respectively (Hartmann et al., 2002). In other difficu lt-to-root tropical species an evaluation of commercially avai lable rooting hormones should be conducted to determine the optimum concentration to achieve the highest rooting percentage. Conclusion Shortages in cutting availability and poor cutting quality exists among shipments to growers, subsequently low quality rooted li ners supplied by domestic propagation firms

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9 has become a concern for growers wishing to pr oduce tropical plants for sale. Examples include poorly rooted or nutrient deficient plants with weak stems and poor lateral branching Propagators of tropical plants need to adopt nutrient management and PGR treatment programs which produce the maximum quantity of high quality cuttings so that growers who receive rooted line rs can profit from the specialized cultivation of the stock plants. Relating shoot and root performance of cuttings from stock plants fertilized at different rates of liquid fertili zers and treated with lateral branching agents is a means of achieving this goal. Additionally identification of optimum rooting hormone concentrations is essential to the efficien t production of high quality rooted liners. Selected Tropical Species To better understand the signi ficance of selected tropical perennials in this review and purpose for which they are being examine d, detailed descriptions of each are as follows: Allamanda schottii Allamanda or Golden Trumpet ( Allamanda schottii L.) is a member of the family Apocynaceae or Milkweed Family which cont ains other important ornamental plants such as flowering vinca ( Catharanthus roseus L.), milkweed ( Asclepias incartata L.), wax plant ( Hoya carnosa L. f.), oleander, frangipani ( Plumeria alba L.), and vinca vine (Judd et al., 2002). Allamanda is characterized by its showy, trumpet-shaped, yellow to purple-red flowers and is na tive to Tropical America (Griffiths, 1994). Depending on species, allamanda may be used as a climbing vine or as a shrub. It performs well in neglected, poor soil and is vigorous enough to withstand co astal conditions (Courtright, 1988; Walker and Hanly, 1996). Certain specie s of allamanda are reported to propagate easily by semi-hardwood cuttings in summer and fall with up to 2,000 mg.L-1 IBA

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10 applied to the cutting base (Hartmann et al., 2002). There is little information published on propagation during other seasons, using othe r combinations of rooting hormones, or stock plant treatments to improve cutting quantity and quality. Bougainvillea glabra Bougainvilleas are members of the family Nyctaginaceae or the Four O’Clock Family, which includes other orna mentals such as four o’clock ( Mirabalis longiflora L.) and sand verbena ( Abronia latifolia Eschsch.) (Judd et al ., 2002). Bougainvillea is native to South America and was originally collect ed by Commerson, a French Botanist from Brazil. Commerson named the plant, because of its wandering growth habit, after Louis Antonyne de Bougainville, a French navigato r known for his journey around the world near the end of the 18th century (Rama Rao, 1976). Bougainvillea are characterized by colorful bracts which exhibit the best flowering performance during cool season months and s horter day lengths in the U.S. The leaves are simple and alternate, but shape and textur e differ in varieties. The plants can be grown as specimen shrubs, hedges, pot crops, or climbing vines. They grow readily in tropical and sub-tropical areas and are seen in diverse locations from Florida to California, India, and Australia (Hacke tt et al., 1972; Rama Rao, 1976; Auld, 1987). Bougainvillea plants are generally propagate d from stem cuttings or air-layerings, but the root systems are extremely fine and fr agile, with low rooting success, even during the cultivated seasons (Chakraverty, 1970). It has been suggest ed that the reason bougainvillea is not more commercially availabl e is due to its difficulty in propagation (Czekalski, 1989). The published literature offers varying in formation on length of stem cuttings and position. Some suggested cutting lengths in clude: 10 to 12 cm (Aldrich and Norcini,

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11 1995; Atzmon et al., 1997) 15 to 17 cm (Auld, 1987); 20 to 25 cm (Chakraverty, 1970; Yadav et al., 1978) and 80 to 100 cm with t op portions removed back to 15 cm of total length (Mudge et al., 1995). Another, perhaps more effective means determining cutting length in bougainvillea is by counting number of nodes. This process has less of a widespread definition of parameters, ra nging from 2 nodes (Auld, 1987), 3 to 4 nodes (Atzmon et al., 1997), or 5 to 9 nodes (Schoe llhorn and Alvarez, 2002). Most sources agree that cuttings should be se vered 0.5 cm below the bottom node. Bougainvillea cuttings should be taken from wood that is mature to the point where it starts to become stiff. If propagating by softwood cuttings night temperatures should be above 12.8 C (55 F), whereas hardwood cuttings root more efficiently when night temperatures are below 12.8 C (55 F) (Schoellhorn and Alvarez, 2002). Cuttings should be provided with rooting s ubstrate temperatures of 30 C (86 F) (Singh et al., 1976). Mandevilla splendens Mandevilla ( Mandevilla splendens Hook.f.), like allamanda, is a member of the Apocynaceae family. It is an evergreen c limbing vine characterized by large pink to white trumpet-shaped flowers appearing mid to late-summer in the U. S. The twining woody tissue should be supported vi a trellis or other structure. Mandevilla is native to Brazil, but is hardy to USDA Hardiness Zone 10 in protected locati ons (Courtright, 1988; Griffiths, 1994). Mandevilla, formerly known as Dipladenia, has been documented as a popular greenhouse plant for over a century in Great Britain (Dress, 1974). Hartmann et al. (2002) suggest propagating Ma ndevilla cultivar Alice DuPont with one node and half of the leaves removed. The stem cutting should then be dipped in 2,500 mg.L-1 IBA and rooted in summer. Other than ‘Alice DuP ont’, there is little published on how to effectively grow and reproduce Ma ndevilla from stem cuttings.

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12 Nerium oleander Oleander is also a member the family Apoc ynaceae. It should be noted that nearly all members of this taxa are poisonous (Judd et al., 2002), therefore care should be taken when harvesting stem cuttings, handling fini shed plants, and planting and maintaining specimens in garden designs. Hands should be washed thoroughly and care should be taken to avoid sap entering the body. Oleander is an evergreen shrub native to the Mediterranean re gions of Southern Europe and North Africa, where it is widely used outside in lands caping designs and as an indoor potted plant (Arias et al., 2001). It has been documente d that oleander first arrived in the U.S. via Spanish settlers in St. Augustine, FL, and it was likely that these plants arrived as stem cuttings (Popenoe, 1976). (Roeber and Reuther., 1982) Oleander is a versatile tropical perennial, hardy to USDA Hardiness Zone 9 and which can be used as a foundation plant, hedge, or as a featured specimen plant. Once established, oleander produces flowers with virtually no main tenance requirements. It tolerates poor soil condi tions and is reportedly salt tolera nt (Walker and Hanly, 1996). It requires a sunny location for northern gardeners, who could use it as a potted plant to be taken indoors during winter months (McCulla, 1973). Oleander is commonly available as an evergreen shrub reaching maturity at 6 m (20 ft), with single, sometimes double, flowers in pink, white, rose, or red. Dwarf vari eties are also availabl e (Courtright, 1988). Propagation has reportedly been successful from cuttings taken from hardwood or softwood during the summer, at lengths of 15.24 cm (6 in), with quick dip applications of 3,000 mg.L-1 IBA (McCulla, 1973; Hartmann et al., 2002). There is limited literature currently available on how to manage the st ock plants of these species and propagation

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13 during other seasons. However, oleander is reportedly not sus ceptible to ethylene branching treatments (Dole and Wilkins, 1999). Tecoma stans Texas Star [ Tecoma stans (L.) Juss.] or ( Bignonia stans L.), is a member of the Bignoniaceae or Trumpet Creeper family, wh ich includes other known ornamentals as catalpa ( Catalpa bignonioides Walter.), sausage tree ( Jacaranda arborea Urban.), and cape honeysuckle ( Tecomaria capensis Spach.) (Judd et al., 2002). Tecoma is a mounding evergreen shrub native to Central Am erica and is charac terized by its funnelshaped yellow flowers throughout summer and ear ly fall in the subtropics (Walker and Hanly, 1996). In Florida, Tecoma is a deci duous shrub or tree hardy to USDA Hardiness Zone 10, that grows to 7.6 m (25 ft) at ma turity, unless pruned, and flowers only for a short period in autumn (Courtright, 1988). Little information is published on how to commercially produce Tecoma or how to ma nage stock plants for improved cutting quantity and quality. Objectives 1. Determine which of three N liquid fertilizer concentrations is optimal for producing the greatest number of bouga invillea cuttings with the highest rooting percentage and quality This study will also simula te long distance shipping, as the cuttings will be harvested in one location an d sent to another for evaluation. 2. Determine which of three concentrations of three commercially available lateral branching PGRs applied to stock plants will most efficiently increase cutting quantity and quality, while not adversely a ffecting subsequent rooting of cuttings. PGRs surveyed will compare three con centrations of chemicals containing cytokinin, ethylene, or gibberellin and cytokinin in combination. 3. Determine the optimum concentration of liquid KIBA, delivered via quick dip, for improving both quantity and quality of adve ntitious root formation in select difficult-to-root tropical perennials. KI BA will also be compared to untreated control and commercial standard quick-di p Dip N’ Grow (Dip N’ Grow Inc., Clackamas, Ore.)(Indole-3-butyric acid 1%, Naphthalene acetic acid 0.5%)

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14 CHAPTER 2 STOCK PLANT NUTRITION Introduction Because the popularity of gardening with tr opical annuals and perennials has grown in the U.S., there has been a considerable increase in production and sales of tropical plants over the past severa l years (Bowden, 2004). Shortage s and poor quality cuttings supplied by domestic propagation firms have become concerns for growers wishing to produce tropical plants for sale. Propagator s have expressed a need for standardized production practices to maximize cutting yi eld and subsequent adventitious root formation in propagules; however, little is published on how to effectively manage tropical stock plants (Hartmann et al., 2002). A tropical species that has potential to be more widely used by gardeners is bougainvillea ( Bougainvillea glabra Choisy). This multi-use plant grows best in U.S. landscapes during summer months followed by at tractive bracts disp layed during natural short-days. Unfortunately because of its warm temperature production requirements and difficulty in propagation, commercial avai lability is limited (Czekalski, 1989). Bougainvillea is generally propagated by stem cuttings or air-layering, yet the rooting percentage is low, even during the cultivat ed seasons (Chakraverty, 1970). The root systems are also known to be extremely fine and fragile (Auld, 1987) Bougainvillea root best from semi-hardwood to hardwood cutti ngs depending on temperature (Schoellhorn, 2002), location (Auld, 1987), and time of year (Chakraverty, 1970). These plants have

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15 been documented to benefit from bottom heat w ith rooting substrate temperatures of 30 C (Singh et al., 1976). Nitrogen (N) concentration is crucial in cut tings as it is important for nucleic acid and protein synthesis in plan t tissue (Hartmann et al., 2002). When considering the role N has in these metabolic processes, one c ould strongly argue its importance in root initiation (Blazich, 1988). Nitrogen concentrat ion also affects cutt ing yield and rooting performance of cuttings. While excessive N may negatively affect cutting propagation (McAvoy, 1995), heavy fertilization in some tr opical plants such as bougainvillea, has been observed to inhibit flowering an d promote growth (Schoellhorn, 2002). Although optimum nutrient levels of tropical stock plants remain undefined, it is crucial that propagules come from a nutritionally healthy source (Hartmann et al., 2002). Limited research has been conducted in the management of stock plants of bougainvillea for purposes of maximizing quantity and quali ty of cuttings suitable for propagation; therefore the experimental objective was to evaluate the impact of three nitrogen concentrations on stock plant yi eld and cutting performance of Bougainvillea ‘Purple Small Leaf’ and ‘Raspberry Ice’. Materials and Methods Experiment 1 Rooted stem cuttings of Bougainvillea glabra ‘Raspberry Ice’ and ‘Purple Small Leaf’ were transplanted, three per pot, into 2.8-L (0.74-gal) [15.2-cm (6-inch diameter)] round plastic containers on 18 April 2005. Th e root substrate was Fafard 2 (Fafard, Anderson, S.C.), which contained 6.5 sphagnum p eat : 2 perlite : 1.5 vermiculite (v/v). A continual liquid fertilization program was in itiated on 23 May, at which time a soft pinch was provided to develop a canopy for harvesting cuttings. Fertiliza tion treatments were

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16 applied with N concentration levels at 100, 200, or 300 mg.L-1. Phosphorous (P), potassium (K), calcium (Ca), and magnesium (Mg) were held constant at 20, 200, 100, and 50 mg.L-1, respectively. Micronutrients were provided using soluble trace element mix in solution (S.T.E.M.) (The Scotts Co mpany, Marysville, Ohio) at a rate of 6.78 mg.L-1. Supplemental iron was applied using fe rric ethylenediaminetertraacetic acid (FeEDTA) (Sequestrine) at a rate of 1.76 mg.L-1. Ammoniacal-nitrogen was 6% to 35% of the total nitrogen in all treatments. Plants were fertigated through a drip irrigation system. The experiment was a randomized co mplete block design with 12 single-plant replications of each of three treatments. Stock plants were grown in a greenhouse under natural photoperiod and irradiance with day/ night minimum setpoint temperatures of 21/18 C. Leachate samples were colle cted initially 4 weeks af ter potting and biweekly thereafter until 28 weeks after potting via a m odified Virginia Tech Extraction Method (VTEM) (Wright, 1986) to dete rmine pH, electrical conduc tivity (EC), and nitrate nitrogen (NO3-N). Distilled water (85 ml) was app lied to the substrate surface to displace 50 ml of leachate. Leachate solutions were analyzed for pH (pHep pH meter; Hanna Instruments, Woonsocket, R.I.), EC (DiS T WP4 EC meter; Hanna Instruments), and NO3-N (Horiba Compact Ion Meter; C-141 Spectrum Technologies, Inc., Plainfield, Ill.). Leachate was collected from 3 random experi mental blocks at each sampling date and values were recorded as re plications 1, 2, and 3, respectiv ely. Photosynthetic photon flux (PPF) was recorded each day at or near solar noon using a Quantum Meter (Spectrum Technologies, East-Plainfield, Ill.). Av erage monthly PPF readings were 453, 586, 426, 596, 647, 355, and 380 molm-2s-1, for June through D ec., respectively.

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17 Stock plants were maintained in Milton, Fla. and cuttings were harvested every three weeks starting 7 weeks af ter potting, on 7 June (H1), 28 June (H2), 19 July (H3), 9 Aug. (H4), 30 Aug. (H5), and 20 Sept. (H6), re spectively. Harvesting protocols consisted of removing all sub-terminal stem cuttings from stock plants by excising the shoot tip 0.5 cm above the first fully mature leaf node, a nd then harvesting the cutting 1 cm below the fourth successive node (Cerveny et al., 2005). The quantity of cuttings was recorded at each harvest. At 7, 13, and 19 weeks after potting shoot length a nd diameter was measured at th e base of 3 randomly selected cuttings per plant. Cutting leaf area was also measured with a LI-COR LI-3100 portable area meter (LI-COR, Lincoln, Nebr.). For tissue analysis, leaves from all cuttings were removed and stock plant blocks 1 to 4, 5 to 8, and 9 to 12 were pooled into treatment replications 1, 2, and 3, respectively. Tissue samples were rinsed in deionized water, then washed in 0.2 N HCl for 30 s, and dried at 70 C for 72 h. Dried tissue was ground in a stainless steel Wiley mill to pass a 1-mm screen (20-mesh). Tissue was then analyzed for macro and micronutrients at Qu ality Analytical Labs (QAL, Panama City, Fla.). Cuttings harvested 10, 16, and 22 WAP were p ackaged in moist towels and inserted into perforated plastic bags. The bags were placed into boxes, along with a temperature and relative humidity sensor (Hobo Pro, m odel number H08-032-08) (Onset Computer, Bourne, Mass.), then stored over-night in a cooler at 10 C. Boxes were sealed and cuttings were then shipped 24h, to simula te industry handling. Upon arrival, boxed cuttings were left intact until the followi ng morning and were then unpacked and planted, totaling approximately 72 h between cutti ng excision and planting (Table 2-1).

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18 Cuttings harvested from stock plant replic ations 1 to 12 of each treatment were pooled and assigned randomly to 6 propagati on replicates with 6 sub-samples of each treatment. Basal portions of cuttings (1 cm ) were inserted into indole-3-butyric acid, potassium salt (KIBA) at 3,000 mg.L-1 for 3 s then planted into 3.3 x 8.8 x 6 cm 6-cell containers in a substrate consisting of 4 pe rlite: 1 vermiculite (v/v) and placed under intermittent mist. Cuttings were maintained under natural photoperiod with day/night set-point temperatures of 20/18 C, and misted daily for 5 s every 20 min (07:00 19:30). Minimum rooting substrate temperatures we re provided at 24 C and bottom heat was supplemented via rooting mats as needed. The experimental design was a completely randomized design with 2 cultivars, 3 N treatme nts, and 6 replications per treatment with 6 sub-samples (pooled) per replicate. Adventitious roots 1.0 mm in length were evalua ted 4 and 6 weeks after planting (WAP). At the first evaluation of each harvest percent survival (based on presence of at least minimal callus formation), rooting qualit y, number of primary roots, number of lateral roots, cutting total dr y weight, and cutting root : s hoot ratio (root weight divided by shoot weight) were recorded. At the s econd evaluation, roots were not counted; all other parameters were recorded in the same manner as the first. Rooting quality was based on a 1 to 5 scale: 1= minimal callus formation with no roots; 2= minimal rooting; 3= moderate, uneven rooting; 4= moderate, uni form rooting; 5= welldeveloped rooting. Cuttings were prepared for dry weight measurement by excision of the rooted portion of stem 1 cm above cutting base. Th en each portion was placed into individual envelopes and dried for 3 d at 70 C. All data were subjected to analysis of variance using

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19 generalized linear model procedure in SAS (SAS institute, Cary, N.C.) and means separated using LSD with P 0.05. Experiment 2 The second experiment was replicated in the same manner as the first experiment, with the following exceptions: Stock plants we re transplanted 18 Ju ly, and given a soft pinch along with initiation of fertilization treatments on 8 Aug. 2005. Cuttings were harvested every three weeks on 23 Aug. (H1), 13 Sept. (H2), 4 Oct. (H3), 25 Oct. (H4), 15 Nov. (H5), and 6 Dec. (H6). Cuttings were handled in the same manner as the first experiment, except they were shipped to Ga inesville, Fla. using Federal Express 24 h service (Table 2-1). Results Experiment 1 Stock Plant Parameters Cutting quantity The harvest date x cultivar and harvest date x N concentration interactions were significant for cutting quantity (Table 2-2). For 'Raspberry Ice', cutting quantity at H1 (10.2) was similar to all other harvest dates. Harvest 4 and H6 were similar to each other with 8.9 and 8.2 cuttings produced per plant, re spectively. Both H4 and H6 were at least 28% less than H2, H3, or H5, all of whic h produced 12.0 cuttings per plant (LSD=2.4, n=36). 'Purple Small Leaf' produced a similar number of cuttings at H1 and H6 (17.0 and 17.4, respectively), while both were at least 30% less than all other harvests. Harvest 5 produced more cuttings (31.3) than H2 (24.8) or H3 (25.1) (LSD=5.2, n=36). Harvest 4 produced a similar number of cuttings to H2, H3, and H5. For all harvests, 'Purple Small Leaf' produced more cuttings per plant, than 'Raspberry Ice' (data not shown). When

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20 considering both cultivars, differences in cutting quantity were not significant amongst treatments for H1, H2, or H4, with 13.5, 18.4, and 18.9 cuttings produced per plant, respectively (Table 2-2). For H3, 126% more cuttings were harvested from stock plants fertilized with N at 300 mgL-1 than from N at 200 or 100 mgL-1. For H5, stock plants fertilized with N at 200 mgL-1 produced a similar number of cuttings to those fertilized with N at 300 mgL-1, while both concentrations produ ced at least 153% more cuttings than N at 100 mgL-1. For H6, N at 300 mgL-1 produced more cuttings than at 100 mgL1, while both treatments were similar to N at 200 mgL-1. For stock plants fertilized with N at 100 or 300 mgL-1, differences in cutting quanti ty were not significant amongst harvests, with 14.4 and 19.7 cuttings produced pe r plant, respectively. For stock plants fertilized with N at 200 mgL-1, the number of cuttings produ ced at H5 was similar to H4 and H2, yet greater th an H1, H3, and H6. Cutting length, stem diameter, and leaf area There was a harvest date x N concentration interaction for cutting length; the main effect for cultivar will be presented separately (Table 2-3). Stock plants of 'Raspberry Ice' and 'Purple Small Leaf' produced similar length cuttings (7.0 cm). For H1 and H5, differences in cutting length were not si gnificant amongst treatments with lengths of 7.0 and 7.5 cm, respectively (Table 2-4). At H 3, stock plants treated with N at 100 or 200 mgL-1 produced cuttings that were at least 112% longer than those treated with N at 300 mgL-1. Over time cuttings harvested from st ock plants treated with N at 100 mgL-1 had longer cuttings at H5 than at H3, while bot h harvest dates were si milar to H1. Stock plants of bougainvillea produ ced cuttings with a mean st em diameter of 0.24 cm and differences were not significant amongst treatments.

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21 There was a cultivar x N concentration inte raction for leaf area (Table 2-3). The harvest date main effects will be presente d separately. Cuttings from H5 (40.5 cm2) had a 114% larger leaf area than H1 (35.3 cm2), and was 116% larger than H3 (30.4 cm2) (LSD=2.1, n=72). Stock plants of 'Raspbe rry Ice' fertilized with N at 100 mgL-1 produced cuttings with similar leaf area to 200 mgL-1 (45.1 cm2), while both concentrations were gr eater than N at 300 mgL-1 (38.9 cm2) (LSD=4.3, n=36). Differences in leaf area were not signifi cant amongst treatments for stock plants of 'Purple Small Leaf'. Rooting Parameters Percent survival, visual rooting quality, and quantity of primary and lateral roots There was a cultivar x N concentration and a harvest date x N concentration interaction for percent survival of planted cuttings (Table 2-5). Differences in percent survival were not significant for 'Raspberry Ice' and 'Purple Small Leaf' (84% and 58%, respectively). 'Raspberry Ice' had a higher perc ent survival than 'Purple Small Leaf' when stock plants were fertiliz ed with N at 100 to 300 mgL-1 (data not shown). When considering both cultivars, cu ttings planted from H2 had higher percent survival from stock plants provided with N at 300 mgL-1 than from concentrations of 100 to 200 mgL1 (Table 2-6). Differences in percent surviv al were not significant amongst treatments for H4 or H6. Nitrogen fe rtilization at 100 mgL-1 produced cuttings with a higher percent survival at H2 or H4 than at H6. Differen ces in percent survival were not significant amongst harvest dates, with 70% survival when stock plants were fertigated with N at 200 mgL-1. Similar to N at 100 mgL-1, stock plants fertigated with N at 300 mgL-1 had a higher percent survival from H2 or H4 than from H6.

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22 There was a harvest date x cultivar x N c oncentration interacti on for rooting quality (Table 2-5). The main effects for evaluation week will be presented separately. Cuttings evaluated 6 WAP had a root quality rating of 2.2, which was higher than those evaluated 4 WAP (2.0) (LSD=0.2, n=108). For H2, diffe rences in rooting quality were not significant amongst treatments for cuttings of 'R aspberry Ice' (Table 2-7). 'Purple Small Leaf', however, had lower rooting qual ity when provided N at 100 or 200 mgL-1 as compared to 300 mgL-1. For H4, differences in rooti ng quality for both 'Raspberry Ice' and 'Purple Small Leaf' were not significant amongst treatments. For H6, 'Raspberry Ice' cuttings planted from stock plants treated with N at 200 mgL-1 had higher rooting quality than from those treated with N at 100 or 300 mgL-1. Differences in rooting quality for 'Purple Small Leaf' cuttings planted from H6 were not significant amongst treatments. When considering the interaction over time 'Raspberry Ice' exhibited a reduction in rooting quality with each ha rvest of stock plants trea ted with N at 100 to 300 mgL-1. Differences in rooting quality for 'Purple Small Leaf' were not significant over time for most treatments, except with N at 300 mgL-1, which had higher rooting quality from H2 than from H4 and H6. 'Raspberry Ice' had hi gher rooting quality than 'Purple Small Leaf' for all treatments and harvests, except for cutt ings planted from stock plants treated with N at 300 mgL-1 and harvested at H2, or any concentration of N at H6, in which the two rooted similarly. There was a harvest date x cultivar and a cultivar x N concentration interaction for number of primary roots (Table 2-8). Fo r cuttings planted from H2, H4, and H6, 'Raspberry Ice' had a greater number of primary roots than 'Purple Small Leaf' (data not shown). Additionally 'Raspberry Ice' cutt ings planted from H2 produced 14.9 primary

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23 roots per cutting, which was at least 470% more than from H4 (3.2) or H6 (1.2) (LSD=2.8, n=18). 'Purple Small Leaf' also had a similar number of primary roots at H4 and H6 (0.3), both of which were less than H2 (2.7) (LSD=1.4, n=18). When comparing cultivars, stock plants of 'Raspberry Ice' fertilized with N at 100 mgL-1 had a greater number of primary roots (5.5), than 'Purple Small Leaf' (1.1) (LSD=3.0, n=18). Similarly, when fertilized with N at 200 mgL-1, stock plants of 'Raspberry Ice' produced cuttings with great er number of primary roots than 'Purple Small Leaf' (data not shown). When fertilized with N at 300 mgL-1 differences between cultivars were not significant, with 3.8 roots produced per cutting. There was a harvest date x cultivar x N concentration interaction for number of lateral roots (Table 2-8). Th e number of lateral roots on cut tings planted from H4 and H6 was not significantly differe nt amongst treatments, with 6.2 and 2.0 lateral roots for 'Raspberry Ice', and with 0.1 and 0.7 lateral r oots for 'Purple Small Leaf', respectively. Cuttings of 'Raspberry Ice' planted from H2 had a greater number of lateral roots when fertilized with N at 200 mgL-1 (37.2) than at 100 mgL-1 (19.7), however both were similar to N at 300 mgL-1 (29.7) (LSD=12.2, n=6). 'Purple Small Leaf' cuttings planted from H2 produced a greater number of lateral roots from stock plants treated with N at 300 mgL-1 (10.4) than either 100 or 200 mgL-1 (2.5 and 0.2, respectively) (LSD=4.1, n=6). Both cultivars produced a similar num ber of lateral roots (1.7) with cuttings planted at H4 from stock plan ts treated with N at 300 mgL-1 (data not shown). Similarly, for cuttings planted from H6, differences betw een cultivars were not significant for all N concentrations. 'Raspberry Ice' cuttings had a greater number of lateral roots than 'Purple

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24 Small Leaf' when planted from stock plants treated with any N concentration from H2 and N at 100 to 200 mgL-1 from H4. Cutting dry weight and root : shoot ratio There was a harvest date x cultivar x N concentration and a harvest date x evaluation week x cultivar inte raction for total cutting dry we ight (Table 2-5). 'Raspberry Ice', cuttings planted from H2 had at least a 17% lower total dry weight when propagated from stock plants treated with N at 100 mgL-1 than at 200 or 300 mgL-1 (Table 2-9). For cuttings planted from either H4 or H6, differe nces in total dry weight were not significant amongst treatments with 165 and 223 mg per cutting, respectively. 'Purple Small Leaf' cuttings propagated at H2 from st ock plants treated with N at 300 mgL-1 had at least 133% greater dry weight than 100 or 200 mgL-1. Differences in total dry weight were not significant amongst N concentrations fo r H4, with 165 mg total dry weight per cutting. From H6, cuttings propagated from stock plants treated with N at 100 to 200 mgL-1 had greater dry weight than at 300 mgL-1. For each N concentration provided and with all harvest dates, 'Raspberry Ice' cutti ngs had greater dry wei ght than 'Purple Small Leaf' when planted from all harvest dates a nd for each N concentration applied to stock plants. 'Raspberry Ice' cuttings evaluated from H2 had a greater dry weight 6 WAP than 4 WAP (257 vs 213 mg, respectively) (LSD=29, n= 18). Similarly, H4 cuttings evaluated 6 WAP had higher dry weight than 4 WAP (data not shown), however at H6 those cuttings evaluated 6 WAP had an 11% lower dr y weight than 4 WAP (235 vs 210 mg, respectively) (LSD=22, n=18). 'Purple Small L eaf' cuttings also had higher dry weight 6 WAP as compared to 4 WAP for those plante d from H2 (160 vs 132 g) (LSD=27, n=18).

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25 Yet from H4 and H6, differences in tota l dry weight were not significant between evaluation weeks, with 101 and 132 mg pr oduced per cutting, respectively. There were harvest date x evaluation week x cultivar, harvest date x evaluation week x concentration, and cultivar x N concen tration interactions for the cutting root : shoot ratio (Table 2-5). 'Raspberry Ice' cu ttings had a higher root : shoot ratio than 'Purple Small Leaf' for each concentration of N applied, however differences in root : shoot ratio were not significant amongst treat ments applied to 'Raspberry Ice' with a mean ratio of 0.26 per cutting. 'Purple Small L eaf' had a higher root : shoot ratio with N at 300 mgL-1 than at 200 mgL-1 (0.18 vs 0.14), while both were similar to 100 mgL-1 (0.15) (LSD=0.04, n=36). While 'Raspberry Ice' had a higher root : shoot ratio than 'Purple Small Leaf' for H2 and H4 at bot h 4 and 6 WAP (data not shown), cuttings planted from H6 had a similar root : shoot ratio between cultivars of 0.15 and 0.14 at 4 and 6 WAP, respectively. When considering both cultivars differe nces in root : shoot ratio were not significant amongst treatments or evaluation weeks for H2, H4, and H6, except cuttings evaluated 6 WAP from H6. Cuttings planted fr om H6 had a greater root : shoot ratio 6 WAP from stock plants treated with N at 200 mgL-1 (0.17) than from N at 300 mgL-1 (0.11), while both were similar to 100 mgL-1 (0.14) (LSD=0.03, n=12). When evaluated 4 WAP, cuttings planted from N concentrations of 100 or 200 mgL-1 showed similar root : shoot ratios amongst harvest dates. When N was applied at 300 mgL-1, cuttings evaluated 4 WAP had a higher root : shoot ratio at H2 than H4 or H6 (data not shown). For cuttings evaluated 6 WAP, those propagated from stock pl ants treated with N at 100 mgL-1 had a higher root : shoot ratio from H2 (0.27) or H4 (0.23) than H6 (0.14)

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26 (LSD=0.09, n=12). Cuttings planted from stock plants fertigated at 300 mgL-1 produced similar root : shoot ratios with H2 or H4 and greater than H6 (d ata not shown), however cuttings planted from stock plants treated with N at 200 mgL-1 were similar amongst harvest dates with a root : shoot ratio of 0.24. Stock plant leachate There was a sampling date x N concentra tion interaction of leachate pH; main effects for cultivar will be presented separate ly. The root substrate of stock plants of 'Raspberry Ice' had a mean leachate pH of 5.3, which was greater than that of 'Purple Small Leaf' (5.1). Generally pH values decr eased with increasing concentrations of N applied to stock plants (Figure 2-1). Vari ations occurred over time and increased concentrations of N applied to stock plants tended to reduce pH va lues below acceptable values of 5.5 to 6.0 (Dole and Wilkins, 2005). Although pH values were in the acceptable range with N at 100 mgL-1, rooting was not improved. The sampling date x N concentration interaction was not significant for electrical conductivity (EC) therefore the main effects will be presented. Overall, EC values remained within the recommended range of 2.0 to 3.5 mS/cm for producing bougainvillea plants established by Whipker et al. (2003) (Figure 2-2). A dec ling then increase in soluble salts was observed during H2 to H4, s uggesting nutrients were utilized for growth of new shoots or leached. As the concentration of N delivered to stock plants increased, so did leachate EC. Stock plants fertigated with N at 300 mgL-1 had the highest EC values of 3.84 mS/cm, while N at 200 mgL-1 (2.92 mS/cm) was greater than 100 mgL-1 (1.87 mS/cm) (LSD=0.56, n=48). When comp aring cultivars, 'Raspberry Ice' (3.24 mS/cm) had higher EC values than 'Purple Small Leaf' (2.52 mS/cm) (LSD=0.46, n=72).

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27 There was a sampling date x cultivar interaction for nitrate nitrogen (NO3-N); main effects for N concentration will be presented separately. Generally NO3-N values were shown to increase with removal of plant tissue during cutting ha rvests (Figure 2-3). This trend was more evident in stock plants of 'Purple Small Leaf' than 'Raspberry Ice', although overall leachate NO3-N values tended to be higher for 'Raspberry Ice'. As N concentration increased, NO3-N values also increased, and were highest from stock plants fertigated with N at 300 mgL-1 (391.9 mgL-1) while N at 200 mgL-1 (278.5 mgL-1) was greater than at 100 mgL-1 (179.8 mgL-1) (LSD=83.9, n=48). Foliar analysis There was a harvest date x cultivar in teraction for foliar N, phosphorous (P), potassium (K), calcium (Ca), and Magnesium (Mg); a harvest date x N concentration interaction for foliar N, P, Ca, and Mg; and a cultivar x N concentration interaction for foliar P, K, and Mg (Table 2-10). 'Raspberry Ice' was generally similar to 'Purple Small Leaf' when comparing foliar N or Mg c oncentrations, although when significant 'Raspberry Ice' tended to be greater than 'P urple Small Leaf' (Table 2-11). For K and P tissue concentrations, 'Purple Small Leaf' tended to be greater than 'Raspberry Ice' at each harvest, while P tissue concentrations tended to be greater for 'Raspberry Ice' than 'Purple Small Leaf'. Generally foliar N concentration was simila r with each concentration of N provided to stock plants, however when cuttings were analyzed from H5, foliar N concentration increased as N concentration increased (Table 2-12). For foliar P, all N fertigation concentrations generally had a similar percentage, except t hose cuttings analyzed from H3, where P levels decreased as N concentr ation increased. Both foliar Ca and Mg concentrations decreased with increasing concen trations of N applied. This decrease may

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28 be due to the low pH substrate inhibiting uptake of these nutrients (W hipker et al., 2003). Magnesium levels remained within acceptable ranges of 0.2% to 0.8% as recommended by Dole and Wilkins (2005); however Ca levels fell below the 1.0% to 2.0% recommendation, when the N concentration in creased to 300 mgL-1 (Table 2-12). When tissue data was compared for each cultivar, both had similar results for P levels (Table 2-13). As the N concentration delivered to stock plan ts increased, foliar K concentration decreased; this trend occurred at lower N concentrations for 'Purple Small Leaf' than for 'Raspberry Ice'. For foliar Mg concentration, 'Raspberry Ice' levels decreased with increased concentrations of N applied, while 'Purple Small Leaf' had higher Mg levels with N at 200 mgL-1 than at 100 or 300 mgL-1. Experiment 2 Stock plant parameters Cutting quantity There was a harvest date x cultivar and a cultivar x N concentration interaction for cutting quantity (Table 2-14). Harvest 3 produc ed the most cuttings of 'Raspberry Ice' at 9.2 per plant, while H1 (4.9), H2 (5.9), H4 (4.6), and H5 (5.4) all produced similar quantities, which were at least 157% more than H6 (2.9) (LSD=1.4, n=36). For 'Purple Small Leaf', cutting quantity was similar at H4 (21.7) and H3 (20.7), both of which were greater than all other harvest dates. Harvest 2 (16.3) produced more cuttings than H5 (12.9), yet both were similar to H6 (15.4). Harvest 1 produced 7.6 cuttings per plant, which was at least 42% less than any othe r harvest (LSD=3.2, n=36). When comparing cultivars, 'Purple Small Leaf' produced cuttings in greater quantity than 'Raspberry Ice' during all harvests (d ata not shown).

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29 'Purple Small Leaf' stock plants produced more cuttings that 'Raspberry Ice' for all N concentrations applied (Table 2-14). 'Raspberry Ice' stock plants produced more cuttings with N at 300 mgL-1 than at 100 or 200 mgL-1. For 'Purple Small Leaf', cutting quantity was greater for stock pl ants treated with N at 200 mgL-1 than at 100 mgL-1 while yield from both treatments was similar to N at 300 mgL-1. Cutting length, stem diameter, and leaf area There was a harvest date x cultivar intera ction for cutting length (Table 2-15). Differences in cutting length were not significant between cultivars for H1 or H3 with 7.7 and 8.2 cm per cutting, respectively. For H 5, cuttings harvested from stock plants of 'Purple Small Leaf' had 116% longer cuttings than from 'Raspberry Ice' (10.2 vs 8.7 cm) (LSD=1.1, n=34). When considering only 'R aspberry Ice', cutting length was longer from H5 (8.7 cm) than H1 (7.9 cm), while H3 was similar to both with 8.4 cm per cutting (LSD=0.5, n=35). 'Purple Small Leaf' had a si milar cutting length from H1 and H3 (7.5 and 8.0 cm, respectively), both of which were less than H5 (10.1 cm) (LSD=0.9, n=36). Differences in stem length for either cultiv ar were not significant amongst treatments. There was a harvest date x N concentration and a harvest date x cultivar interaction for stem diameter (Table 2-15). For H1 'Purple Small Leaf' had a 16% smaller stem diameter than 'Raspberry Ice' (0.19 vs 0.23 cm, respectively). Both cultivars had similar stem diameter at H3 (data not shown). At H5, 'Purple Small Leaf' had larger sized stems (0.24 cm) than 'Raspberry Ice' (0.20 cm) (LSD=0.02, n=34). 'Raspberry Ice' cuttings from H3 had a stem diameter of 0.24 cm, which was 108% and 115% larger than H1 (0.23) and H5 (0.20), respectively (LSD=0.01, n= 35). Alternatively, 'Purple Small Leaf' cuttings had a larger stem diameter at H5 than H3 (0.24 vs 0.21 cm), both of which were greater than H1 (0.19 cm) (LSD=0.02, n=36). Although statistical differences were

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30 shown, differences in stem diameter of 0.03 cm may not be considered commercially important for either cultivar. When consid ering both cultivars, differences in stem diameter were not significant amongst treatment s for any harvest date. When considering harvest dates within treatments, cuttings harv ested from stock plants treated with N at 300 mgL-1 had a larger stem diameter from H3 (0.23 cm) than either H5 or H1, which had similar a diameter of 0.20 cm (LSD=0.02, n=24). Differences in stem diameter amongst harvest dates were not significant for other N concentrations. There was a harvest date x cultivar and a cultivar x N concentration interaction for leaf area (Table 2-15). From H1, cuttings of 'Raspberry Ice' had a 157% larger leaf area than 'Purple Small Leaf' (76.5 vs 48.5 cm2, respectively) (LSD=4.9, n=36). Similarly, at H3, 'Raspberry Ice' had a leaf area of 47.1 cm2, which was 115% larger than that of 'Purple Small Leaf' (40.8 cm2) (LSD=4.3, n=36). At H5, however, 'Purple Small Leaf' had larger leaf area than 'Raspberry Ice' (31.6 vs 24.1 cm2) (LSD=3.9, n=36). When considering differences in 'Raspberry Ice' am ongst harvest dates, leaf area from H1 (76.5 cm2) was larger than H3, both (47.1 cm2) of which were larger than H5 (24.1 cm2) (LSD=4.7, n=35). 'Purple Small Leaf' had larger leaf area at H5 (10.1 cm2) than at H1 or H3 (7.5 or 8.0 cm2, respectively) (LSD=0.9, n=36). Differences for both 'Raspberry Ice' and 'P urple Small Leaf' were not significant amongst treatments with a leaf area of 51.0 and 40.3 cm2, respectively (Table 2-16). When comparing cultivars, stock plants fertigated with N at 100 to 200 mgL-1, had a larger leaf area with 'Raspberry Ice' than 'P urple Small Leaf'. Differences in leaf area were not significant between cultivars when stock plants were provided N at 300 mgL-1.

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31 Rooting parameters Percent survival, visual rooting quality, and quantity of primary and lateral roots There was a harvest date x evaluation w eek x N concentration interaction for percent survival (Table 2-17). Main effects for cultivar differences will be presented separately. Percent survival of 'Raspberry Ice' was 93%, while 71% of 'Purple Small Leaf' cuttings survived (LSD=5, n=90). Di fferences in percent survival were not significant amongst treatments for cuttings plante d at H2, H4, or H6 and evaluated 4 or 6 WAP, respectively (data not shown). When considering N concentration, stock plants fertilized with N at 300 mgL-1 produced cuttings at H4 that had a higher percent survival 6 WAP than at 4 WAP (86% vs 55%) (LSD =20, n=12), otherwise all other harvest plantings had similar survival percentage s at both 4 and 6 WAP, regardless of N concentration. There was a harvest date x cultivar x N concentration and a harvest date x evaluation week interaction fo r rooting quality (Table 2-17 ). When considering both cultivars, cuttings evaluated 6 WAP had a higher rooting qual ity than those evaluated 4 WAP from H2, H4, and H6 (dat a not shown). For both 'Raspberry Ice' and 'Purple Small Leaf', rooting quality was similar amongst treatments and harvests. One exception was cuttings of 'Purple Small Leaf' planted fr om stock plants treated with N at 100 mgL-1, which had a lower root quality rating than those provided with N at 200 or 300 mgL-1 (Table 2-18). 'Raspberry Ice' when harv ested from N concentrations of 100 mgL-1, produced better quality rooted cuttings when planted from H2 and H6, than from H4. Nitrogen at 200 mgL-1 produced variable results in r ooting quality from each harvest, with mean ratings of 3.9, 1.6, and 3.0 from H2, H4, and H6, respectively. Similarly, cuttings planted from H2 had higher rooting quality than H6, whic h was higher than H4

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32 with N at 300 mgL-1. While 'Purple Small Leaf' cuttings had similar rooting quality from H4 and H6, rooting quality of cu ttings planted from N at 100 mgL-1 at H2 was higher. Differences in rooting qual ity were not significant amongst harvest dates with N concentrations from 200 to 300 mgL-1. There was a harvest date x cultivar x N concentration interaction for number of primary roots (Table 2-19). For most harves t plantings, both 'Raspberry Ice' and 'Purple Small Leaf' had a similar num ber of primary roots amongst treatments. However, 'Raspberry Ice' from H2 had more primary root s per cutting when fertilized with N at 200 mgL-1, than at 300 mgL-1, and both of these treatments were similar to N at 100 mgL-1. Cuttings of 'Purple Small Leaf' planted at H6 had 281% more primary roots when stock plants were fertigated with N at 300 mgL-1 than at 100 mgL-1, while both treatments were similar to N at 200 mgL-1. For cuttings planted from H2, 'Raspberry Ice' had 264%, 425%, and 237% more primary roots than 'Purple Small Leaf' when stock plants were fertigated with N at 100, 200, and 300 mgL-1, respectively (data not shown). 'Raspberry Ice' and 'Purple Small Leaf' cuttings both had a similar number of primary roots amongst the three N concentrations when planted from H4 and H6. There was a harvest date x cultivar inte raction for number of lateral roots. 'Raspberry Ice' had 18.5 lateral roots per cut ting for H2, which was at least 506% more than H4 or H6 (1.4 or 3.7, respectively) (L SD=5.6, n=12). 'Purple Small Leaf' had more lateral roots per cutting when planted from H6 (6.5) than H2 (3.4) or H4 (1.4) (LSD=2.6, n=18). For cuttings planted from H2, 'Pur ple Small Leaf' had an 82% lower number of lateral roots than cuttings of 'Raspberry Ice' (data not shown). 'Raspberry Ice' and 'Purple

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33 Small Leaf' had a similar numbe r of lateral roots from H4 and H6, with 1.4 and 5.1 lateral roots per cutting, respectively. Cutting dry weight and root : shoot ratio There was a harvest date x evaluation week and a harvest date x cultivar interaction for cutting total dry weight (Table 2-17). Fo r cuttings planted from H2, those evaluated 6 WAP had greater total dry weight than at 4 WAP (291 vs 231 mg) (LSD=39, n=35). For H4 and H6, differences in total dry weight were not significant between evaluation weeks with 219 and 173 mg per cutting, respectively. When comparing cultivars, 'Raspberry Ice' and 'Purple Small Leaf' had similar cutti ng dry weight when planted from H2 and H6 with 244 and 253 mg per cutting, respectively. For H4, 'Raspberry Ice' cuttings had greater dry weight than 'Purple Small Leaf' (245 vs 211 mg) (LSD=29, n=22). Dry weight of 'Raspberry Ice' cuttings decr eased over time with 289, 245, and 163 mg per cutting from H2, H4, and H6, respectivel y (LSD=32, n=16). 'Purple Small Leaf' responded similarly over time with 251, 211, and 177 g per cutting from H2, H4, and H6, respectively (LSD=32, n=45). There was a harvest date x evaluation week and a harvest date x cultivar interaction for the cutting root : shoot ratio (Table 2-17) The root : shoot ra tio was similar between 'Raspberry Ice' and 'Purple Small Leaf' at H 2, H4 and H6. 'Raspberry Ice' cuttings had similar root : shoot ratios for H2 and H6 with 0.24, which was greater than H4 (0.15) (LSD=0.05, n=16). 'Purple Small Leaf' cuttings performed similarly, with H2 (0.24) or H6 (0.28) being greater than H4 (0.15) (L SD=0.05, n=45). When considering evaluation weeks, cuttings evaluated from either H4 (0.15) or H6 (0.26) had similar root : shoot ratios at both 4 and 6 WAP, while cuttings evaluated 4 WAP from H2 had a 42% lower root : shoot ratio than at 6 WA P (0.17 vs 0.32) (LSD=0.05, n=34).

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34 Stock plant leachate There was a sampling date x N concentr ation interaction for leachate pH, differences in cultivar were not significant. Over time substrate pH increased with N at 100 mgL-1, while a decrease in pH was obs erved with N at 200 to 300 mgL-1 (Figure 24). The sampling date x N concentration inte raction was not significant for leachate EC, therefore the main effects will be presen ted separately. Both cultivars had similar EC values. Over time, values remained within the recommended range of 2.0 to 3.5 mS/cm for producing bougainvillea plants (Fig ure 2-5). As the concentration of N applied to stock plants increased, EC also increased. Nitrogen at 300 mgL-1 had the highest leachate EC (3.62 mScm-1), while 200 mgL-1 (2.89 mScm-1) was higher than 100 mgL-1 (1.76 mScm-1) (LSD=0.32, n=48). There was a sampling date x N c oncentration interaction for NO3-N, differences in cultivar were not significant. In general, leachate values for NO3-N were shown to increase with removal of plant tissue at cutti ng harvests when stock pl ants were fertigated with N at 200 to 300 mgL-1 (Figure 2-6). Leachate NO3-N values were excessively high with N at 300 mgL-1. Foliar analysis There was a harvest date x cultivar x N c oncentration interacti on for foliar Ca and Mg concentrations. There was a harvest x cultivar interaction for foliar P and K concentrations. There was a harvest x N con centration interaction for foliar N, while main effects for cultivar will be presented separately (Table 2-20). 'Raspberry Ice' had greater foliar N concentration than 'Pur ple Small Leaf' (4.59% vs 4.43%) (LSD=0.14, n=18). Early in the season, at H1, stoc k plants fertigated with N at 200 mgL-1 (4.52%)

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35 had greater foliar N concen tration than at 300 mgL-1 (4.16%), while both concentrations were similar to N at 100 mgL-1 (4.30%) (LSD=0.27, n=6). During H3, foliar N concentration was similar for stock plan ts fertigated with N at 200 to 300 mgL-1 (4.94%), while both were greater than 100 mgL-1 (4.14%) (LSD=0.33, n=6). Harvest 5 followed a similar pattern, as the N concentration delivered to stock plants increased, N concentration in the foliage also increased (d ata not shown). When considering foliar K concentrations, cuttings harvested from stock plants treated with N at 100 mgL-1 had the highest tissue concentrati ons (4.16%), while 200 mgL-1 (3.93%) was greater than 300 mgL-1 (3.54%) (LSD=0.16, n= 27). 'Purple Small Leaf' had a similar K concentration in foliage to 'Raspberry Ice' during H1 and H5, while at H3 'Purple Small Leaf' (4.23%) had a greater K concentration than 'Raspberry Ice' (3.63%) (LSD=0.31, n=9). Phosphorous levels were similar for all treatments duri ng the second study. During H1, 'Purple Small Leaf' cuttings had a greater foliar P concentr ation than 'Raspberry Ice' (0.52% vs 0.42%) (LSD=0.05, n=9). The third harvest followed an opposite trend, with P being greater in 'Raspberry Ice' (0.32%), than 'Purple Sma ll Leaf' (0.29%) (LSD=0.02, n=9). Differences in cultivar responses were not significant at H5, with 0.26% P measured in tissue. Over time, foliar P and K concentrations decrease d for both cultivars, although values were still within the acceptable range of 0.2% to 0.8% for P, and 3.0 to 5.5% for K (Whipker et al., 2003). When considering foliar Ca levels observed for 'Raspberry Ice' and 'Purple Small Leaf', concentrations were generally similar amongst N c oncentrations applied. During H3 however, decreased Ca concentrations we re observed as N con centration increased (Table 2-21). As the season progressed, 'Raspbe rry Ice' exhibited a decline in foliar Ca.

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36 Although the characteris tic leaf distortions and chlorosi s commonly associated with Ca deficiency were not observed, resultant slow growth due to a minor calcium deficiency may have contributed to the re duction in cutting quantity ha rvested later in the season (Whipker et al., 2003). Magnesium levels were shown to be above the recommended range of 0.2% to 0.8% during H1 for both cultivars (Table 2-21). Decreased foliar Mg was observed as the season progressed for both cultivars, however values remained within the recommended range. Discussion During the first experiment, 'Purple Sma ll Leaf' produced more cuttings than 'Raspberry Ice'. Although cutting length, stem diameter, and leaf area were similar between cultivars, percent survival, rooting qua lity, number of lateral roots, total cutting dry weight, and root : shoot ra tios for planted cuttings were shown to be greater with 'Raspberry Ice' than 'Purple Small Leaf'. When considering both cultivars, stock plan ts fertigated with N concentrations of 200 to 300 mgL-1, produced greater quantities of cuttings, when compared to 100 mgL-1. At times, all treatments produced similar numb ers of cuttings, yet throughout the study N at 300 mgL-1, more consistently produced the largest yield (Table 2-2). Additionally, when comparing consistency of harvest dates, stock plants fertigat ed with N at 100 or 300 mgL-1 produced similar numbers of cuttings at each harvest, while results from N at 200 mgL-1 varied. 'Purple Small Leaf' produced cuttings with a shorter stem length than 'Raspberry Ice', however the 0.3 cm difference may not be considered commercially important. When considering N concentration, cutti ng length was generally similar amongst treatments, however at the third harvest st ock plants fertigated with N at 100 to 200

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37 mgL-1 produced longer cuttings than at 300 mgL-1. Cutting leaf area was generally similar for all treatments, although 'Raspberry Ice' showed a reduction in area as N concentration increased. 'Purple Small Leaf' had a similar leaf area for all N concentrations provided. As the N concentr ation increased later in the harvesting season, a slight trend was shown with both cultivars toward shorter cu ttings with decreased leaf area. Percent survival for cuttings planted from H2 was higher with cuttings planted from stock plants treated with N at 300 mgL-1 than from concentrations of 100 to 200 mgL-1, otherwise all treatments had similar su rvival. When comparing harvest dates, cuttings planted from H6 had a considerable reduction in survival as compared to the other two plantings. Improved su rvivability with N at 200 mgL-1 than 300 mgL-1, occurred with cuttings planted from H6, ther efore growers may want to consider reducing fertilizer concentrations as the growing s eason progresses into the cool season. Rooting quality of 'Purple Small Leaf' cuttings was improved with N at 300 mgL-1, as compared to lower concentrations early in the st udy, yet as the season progressed treatment differences were not significant. 'Raspberry Ice' cutting performance was generally not influenced by treatment early in the season, yet had better rooting quality when planted from H6 when N was applied at 200 mgL-1, as compared to 100 or 300 mgL-1. When comparing primary root number all N concentrations produ ced a similar quantity. In general both cultivars had a reduction in prim ary roots as the season progressed; this trend seems to coincide with the decreases in visual rooting quality shown during the same time period. For 'Raspberry Ice', this reduction in rooting quality at H6 may be related to the reduction in both stem le ngth and leaf area seen with increased

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38 concentrations of nitrogen app lied to stock plants later in the season. Lateral root number was generally improved with N concentrations of 200 to 300 mgL-1 when applied to stock plants of 'Raspberry Ice', and with 300 mgL-1 applied to 'Purple Small Leaf'. Differences in total cutting dry weight and the root : shoot ratio seem to coincide with other rooting parameters previously discussed, as 'Purple Small Leaf' was shown to have increased total cutting dry weight and an improved root : shoot ratio with N at 300 mgL-1 as compared to the lower concentra tions applied to stock plants. Although 'Raspberry Ice' was not significant amongst trea tments for the root : shoot ratio, total cutting dry weight was great er from N at 200 to 300 mgL-1 than at 100 mgL-1. When comparing all parameters, stock plan ts of 'Raspberry Ice' and 'Purple Small Leaf' were shown to achieve the greatest benefits in yi eld and subsequent rooting performance when stock plants were fertigated with N at 200 to 300 mgL-1 during Expt. 1 (warm season). Results were similar to studies conducted during the 2005 spring growing season, in which the recommended concentration was 200 mgL-1 (Cerveny et al., 2005). During the second experiment, 'Purple Small Leaf' produced more cuttings than 'Raspberry Ice'. When comparing treatment s, 'Raspberry Ice' produced the greatest quantity of cuttings when stock plants were fertigated with N at 300 mgL-1 as compared to 100 to 200 mgL-1. Alternatively, 'Purple Small Leaf’ stock plants were shown to have optimum yield with N at 200 mgL-1 as compared to 100 or 300 mgL-1, suggesting no benefit to the increas ed concentration. Cutting length was generally similar for bot h cultivars, except 'Purple Small Leaf' produced longer cuttings than 'Raspberry Ice' during H5. 'Raspberry Ice' generally had a

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39 larger leaf area than 'Purpl e Small Leaf' with lower con centrations of N applied, yet differences were not significant as concentrations increased to 300 mgL-1. When comparing harvests, leaf area tended to be grea ter with 'Raspberry Ice' than 'Purple Small Leaf' during earlier harvests, although at H5 'Purple Small Leaf' had a larger leaf area than 'Raspberry Ice'. As the season progresse d into the cooler months, 'Raspberry Ice' stock plants produced fewer cu ttings with shorter lengths and less leaf area; this was shown to a lesser degree with 'Purple Small Leaf'. This change in performance might suggest a seasonal effect, although a similar tr end was observed to a lesser degree during summer. Because bougainvillea flowers under na tural short days, flower initiation may have inhibited vegetative growth (Chakr averty, 1970). Another possibility for the observed changes in cutting yield may be due to a natural reduction in vigor associated with 18 to 20 weeks in production. In comme rcial floriculture, stock plants of other species are often replanted several times per year to avoid reductions in vigor (Carlos Villalobos, General Manager, Ball Linda Vista facility, Costa Rica, personal communication). Alternatively, the foliar analys is data which showed slight Ca and Mg deficiencies, coupled with reduced pH valu es observed in stock plant leachate may indicate reductions in vigor ar e related to elemental deficien cies associated with low pH. Percent survival was genera lly not influenced by N con centration applied to stock plants and 'Raspberry Ice' tended to be simila r in percentage survival to 'Purple Small Leaf' for most harvest plantings and evaluati on dates. Visual rooting quality was also generally similar amongst N concentration pr ovided to stock plants, except for 'Purple Small Leaf' later in the season, in which higher rate s of N at 200 to 300 mgL-1 provided to stock plants had greater r ooting quality than at 100 mgL-1. Throughout the second

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40 study N fertilization provided to 'P urple Small Leaf' at 200 to 300 mgL-1 produced similar results when planted from all harves ts. 'Raspberry Ice' tended to have better rooting earlier in the study for all treatments, but was generally similar amongst all N concentrations. For the N concentrations investigated, 200 and 300 mgL-1 produced the highest quantity of primary roots for 'Raspberry Ice' and 'Purple Small Leaf', respectively. For lateral roots, differences in treatment were not significant, yet 'Raspberry Ice' was shown to have improved lateral rooting earlier in the season, while 'Purple Small Leaf' performed better as the season progressed. 'Raspberry Ice' wa s greater than 'Purple Small Leaf' when considering number of lateral roots when cutt ings were planted from H2, although H4 and H6 produced similar results between cultivars. 'Raspberry Ice' and 'Purple Small Leaf' generally had similar total cutting dry weight values throughout the study with few exceptions. Total cutting dry weight of 'Raspberry Ice' decreased with each harvest planting, while 'Purple Small Leaf' values were constant over time. This data furthe r illustrates differences observed with other parameters previously mentioned. The root : shoot ratios were similar for 'Raspberry Ice' and 'Purple Small Leaf' cuttings, and both follo wed a similar trend with harvest number. Cutting root : shoot ratios were generally si milar with each harvest, however a reduction was seen when cuttings we re planted from H4. When considering both studies, cuttings evaluated 4 WAP tended to have similar survival to those evaluated 6 WAP, this sugge sts that although rooti ng may not be at the desired level, no benefit was shown in allo wing cuttings to remain under mist for six weeks. All other parameters that showed improvement 6 WAP as compared to 4 WAP,

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41 such as rooting quality, root number, and dr y weight, all suggest growth with time and would not be considered the result of prolonge d misting. Growers will want to consider removing cuttings from the propagation environm ent when the majority of cuttings have begun rooting to encourage better establis hment of rooted liners. Length of time necessary to propagate bougainvillea cuttings va ries, so growers will want to experiment with each cultivar to determine the ideal harvesting window. Conclusion During the summer study stock plants of bougainvillea were shown to benefit from N at 300 mgL-1. The trend during the second study suggests that 300 mgL-1 may be more appropriate earlier in the growing season, but as the season progresses fertilization program should be reduced to 200 mgL-1. Stock plant leachate pH, EC, and NO3-N should be monitored regularly to assu re optimum root zone management. Bougainvillea glabra 'Raspberry Ice' and 'Purple Small Leaf' were also shown to survive similarly at 4 and 6 WAP under Florida condi tions. Growers should compare the risk of extended propagation time to the benefits achieved in rooting performance for their specific growing environments. Since the response of 'Purple Small Leaf' and 'Raspberry Ice' varied over time and between each other, further studies should be conducted on other cultivars of bougainvillea as well as other tropical spec ies to determine optimum concentrations of N for a successful stock plant management program.

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42 Table 2-1. Temperature and relative humidity data (maximum, minimum, and mean) as recorded during simulated and actual shipping (excision to planting) of cuttings from Harvests (H) 2, 4, and 6 of Expts. 1 and 2, respectively. Temperature (C) Relative Humidity (%) Expt 1 Max. Min. Mean Max. Min. Mean H2 34.4 13.3 22.4 94.5 46.4 81.4 H4 36.1 20.6 26.4 82.5 56.2 69.1 H6 38.8 22.9 26.5 91.7 36.3 72.3 Avg 36.4 18.9 25.1 89.6 46.3 74.3 Temperature Relative Humidity Expt 2 Max. Min. Mean Max. Min. Mean H2 38.8 21.0 25.8 82.1 30.4 65.1 H4 26.0 3.3 17.3 96.7 26.6 63.7 H6 25.6 3.3 13.9 88.0 30.4 72.7 Avg 30.1 9.2 19.0 88.9 29.1 67.2 Table 2-2. Influence of harvest date, cultivar, and nitrogen (N) concentration on quantity of sub-terminal bougainvillea stem cuttings harvested during the warm season (Expt. 1). N Concn Cutting quantity (mgL-1) H1 H2 H3 H4 H5 H6 LSD 100 13.5 16.1 16 15.9 15.6 9.5 NS 200 14 19 17.6 20 25.5 11.4 7 300 13.2 20.1 22.1 20.8 23.9 18.1 NS Significance NS NS *** NS * LSD 3.6 6.6 6.9 Source of variance df Pr > F Rep (R ) 11 0.0061 Harvest date (H) 5 0.0001 Cultivar (C ) 1 0.0001 H x C 5 0.0001 N concn (T) 2 0.0001 H x T 10 0.0495 C x T 2 NS H x C x T 10 NS NS, *, *** Nonsignificant or significant at P 0.05 or 0.001, respectively.

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43 Table 2-3. Significance levels for effects of harvest date, cultivar, and nitrogen (N) concentration on length and leaf area of cuttings harvested from stock plants of bougainvillea (Expt 1). Source df Cutting length Leaf area Rep (R ) 11 NS NS Harvest date (H) 2 0.0001 0.0001 Cultivar (C ) 1 0.0254 0.0001 H x C 2 NS NS N concn (T) 2 NS 0.0018 H x T 4 0.0454 NS C x T 2 NS 0.0063 H x C x T 4 NS NS NSNonsignificant at P 0.05. Table 2-4. Harvest date (H) by nitrogen (N) concentration interac tion for length of subterminal bougainvillea stem cuttings fr om harvested during the warm season (Expt 1). Cutting length (cm) N Concn (mgL-1) H1 H3 H5 LSD 100 7.3 6.7 7.5 0.6 200 6.7 6.7 7.3 0.5 300 6.9 6 7.6 *** 0.6 Significance NS NS LSD 0.5 NS, *, *** Nonsignificant or significant at P 0.05 or 0.001, respectively.

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44 Table 2-5. Significance levels for effects of harvest date, evaluation date, cultivar, and nitrogen concentration provided to stoc k plants on percent survival, visual rooting quality (1=poor; 5=best), tota l dry weight (shoot weight + root weight), and root : shoot ratio of s ub-terminal bougainvillea stem cuttings planted during the warm season (Expt. 1). Source df Percent survival Rooting quality Total dry weight Root : shoot ratio Rep (R ) 5 NS NS 0.0248 NS Harvest date (H) 2 0.0001 0.0001 0.0001 0.0001 Evaluation date (W) 1 NS 0.0185 0.0001 0.0001 H x W 2 NS NS 0.0001 0.0019 Cultivar (C ) 1 0.0001 0.0001 0.0001 0.0001 H x C 2 NS 0.0001 0.0041 0.0001 W x C 1 NS NS NS 0.0015 H x W x C 2 NS NS 0.0001 0.0482 N Concn (T) 2 NS NS 0.0001 NS H x T 4 0.0051 0.0005 0.0001 0.0011 W x T 2 NS NS NS NS H x W x T 4 NS NS NS 0.0314 C x T 2 0.0227 0.0003 NS 0.0096 H x C x T 4 NS 0.0482 0.006 NS W x C x T 2 NS NS NS NS H x W x C x T 4 NS NS NS NS NSNonsignificant at P 0.05. Table 2-6. Harvest date (H) by nitrogen (N) concentration interaction for percent survival of sub-terminal bougainvillea st em cuttings planted from stock plants treated with N at 100, 200, or 300 mgL-1 during the warm season (Expt. 1). Percent survival (%) N Concn (mgL-1) H2 H4 H6 LSD 100 75 77 58 15 200 76 69 63 NS 300 90 80 49 *** 14 Significance NS NS LSD 12 NS, *, *** Nonsignificant or significant at P 0.05 or 0.001, respectively.

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45 Table 2-7. Harvest date (H) by cultivar by N concentration interaction for visual rooting quality (1=poor; 5=best) of sub-termin al bougainvillea stem cuttings planted during the warm season (Expt. 1). Rooting quality N Concn Raspberry Ice Purple Small Leaf (mgL-1) H2 H4 H6 H2 H4 H6 100 3.6 2.8 1.5 1.8 1.3 1.6 200 3.9 2.8 2 1.5 1.1 1.3 300 3.5 2.2 1.3 2.9 1.4 1.1 Significance NS NS * NS NS LSD 0.5 0.8 NS, Nonsignificant or significant at P 0.05, respectively. Table 2-8. Significance levels for effect s of harvest date, cultivar, and nitrogen concentration provided to stock plants on number of primary and lateral roots on sub-terminal bougainvillea stem cuttings planted during the warm season (Expt. 1). Source df Primary roots Lateral roots Rep (R ) 5 NS NS Harvest date (H) 2 0.0001 0.0001 Cultivar (C ) 1 0.0001 0.0001 H x C 2 0.0001 0.0001 N Concn (T) 2 NS 0.0129 H x T 4 NS 0.0094 C x T 2 0.0343 0.0003 H x C x T 4 NS 0.0278 NSNonsignificant at P 0.05. Table 2-9. Harvest date by cultivar by N con centration interaction for total cutting dry weight (shoot weight + root weight) of sub-te rminal bougainvillea stem cuttings planted during the warm season (Expt. 1). Total cutting dry weight (mg) N Concn Raspberry Ice Purple Small Leaf (mgL-1) H2 H4 H6 H2 H4 H6 100 204 152 216 116 98 145 200 253 164 230 138 101 139 300 246 179 222 184 105 111 Significance NS NS *** NS LSD 36 25 23 NS, *, *** Nonsignificant or significant at P 0.05 or 0.001, respectively.

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46 Table 2-10. Significance levels for effects of harvest date, cultivar, and nitrogen (N) concentration on foliar concentration of macronutrients N, phosphorous (P), potassium (K), calcium (Ca), and magnesium (Mg) of sub-terminal bougainvillea stem cuttings harvested dur ing the warm season (Expt. 1). Source df N P K Ca Mg Rep 2 NS NS NS NS NS Harvest date (H) 2 NS 0.0001 0.0001 0.0001 0.0001 Cultivar (C) 1 NS 0.0001 0.0001 0.0001 NS H x C 2 0.0396 0.0038 0.0114 0.0001 0.0001 N concn (T) 2 0.0001 0.0368 0.0001 0.0001 0.0001 H x T 4 0.0213 0.0028 NS 0.0001 0.0002 C x T 2 NS 0.0009 0.0040 NS 0.0001 H x C x T 4 NS NS NS NS NS NSNonsignificant at P 0.05. Table 2-11. Harvest date (H) by cultivar interaction for fo liar nutrient concentration of macronutrients nitrogen (N), phosphorous (P), potassium (K), calcium (Ca), and magnesium (Mg) of sub-terminal bougainvillea stem cuttings harvested during the warm season (Expt. 1). Foliar concn (%) Cultivar HarvestN P K Ca Mg Raspberry Ice H1 4.63 0.40 3.92 0.96 0.62 Purple Small Leaf 4.57 0.37 4.65 1.16 0.52 NS *** *** ** Raspberry Ice H3 5.02 0.33 3.94 0.82 0.55 Purple Small Leaf 4.59 0.33 4.31 1.14 0.60 NS *** NS Raspberry Ice H5 4.80 0.33 3.75 0.83 0.51 Purple Small Leaf 4.87 0.29 4.21 0.87 0.51 NS *** *** NS NS NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively.

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47 Table 2-12. Harvest date (H) by nitrogen (N) concentration interac tion for foliar nutrient concentrations of macronutrients N, phosphorous (P), calcium (Ca), and magnesium (Mg) of sub-terminal bouga invillea stem cuttings harvested during the warm season (Expt. 1).. N concn Foliar concn (%) Harvest (mgL-1) N P Ca Mg H1 100 4.52 0.39 1.12a 0.61 a 200 4.68 0.38 1.11a 0.59 a 300 4.60 0.40 0.96b 0.51 b NS NS ** H3 100 4.56 0.31 b 1.11a 0.65 a 200 4.81 0.32 b 1.06a 0.62 a 300 5.05 0.35 a 0.77b 0.46 b NS *** *** H5 100 4.36 cz 0.31 1.13a 0.63 a 200 4.84 b 0.32 0.85b 0.52 b 300 5.31 a 0.30 0.57c 0.39 c *** NS *** *** NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. z Any two means within a column no t followed by the same letter are statistically different using LSD at P 0.05. Table 2-13. Cultivar by nitrogen (N) concen tration interaction for foliar nutrient concentration of macronutrients pho sphorous (P), potassium (K), and magnesium (Mg) of sub-terminal bouga invillea stem cuttings harvested during the warm season (Expt. 1). Foliar concn (%) N concn (mgL-1) P K Mg Raspberry Ice 100 0.35 4.00a 0.62 a 200 0.36 3.94a 0.55 b 300 0.35 3.66b 0.51 c NS *** *** Purple Small Leaf 100 0.33 4.62a 0.52 b 200 0.32 4.22b 0.60 a 300 0.35 4.32b 0.51 b NS ** *** NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. z Any two means within a column not followe d by the same letter are statistically different using LSD at P 0.05.

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48 Table 2-14. Influence of harvest date, cu ltivar, and nitrogen (N) concentration on quantity of sub-terminal bougainvillea st em cuttings harvested during the cool season (Expt. 2). N concn Cutting quantity (mgL-1) Raspberry Ice Purple Small Leaf LSD 100 4.9 13.9 *** 2 200 4.9 17.5 *** 2.3 300 6.6 15.8 *** 1.8 Significance ** LSD 1 2.2 Source of variance df Pr > F Rep (R ) 11 0.0241 Harvest date (H) 5 0.0001 Cultivar (C ) 1 0.0001 H x C 5 0.0001 N concn (T) 2 0.0037 H x T 10 NS C x T 2 0.006 H x C x T 10 NS NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, 0.001, respectively. Table 2-15. Significance levels for effects of harvest date, cultivar, and nitrogen (N) concentration on length, stem diameter and leaf area of sub-terminal bougainvillea stem cuttings harvested during the cool season (Expt. 2). Source df Cutting length Stem diameter Leaf area Rep (R ) 11 NS NS 0.0051 Harvest date (H) 2 0.0001 0.0044 0.0001 Cultivar (C ) 1 NS 0.0221 0.0001 H x C 2 0.0012 0.0001 0.0001 N concn (T) 2 NS NS NS H x T 4 NS 0.0283 NS C x T 2 NS NS 0.0002 H x C x T 4 NS NS NS NSNonsignificant at P 0.05.

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49 Table 2-16. Cultivar by nitrogen (N) concen tration interaction for leaf area of subterminal bougainvillea stem cuttings ha rvested during the cool season (Expt. 2). N concn Leaf area (cm2) (mgL-1) Raspberry Ice Purple Small Leaf LSD 100 50.6 37.6 9.1 200 53.5 40.1 9.3 300 45.8 43.2 NS Significance NS NS NS, Nonsignificant or significant at P 0.05, respectively. Table 2-17. Significance levels for effects of harvest date, evaluation date, cultivar, and nitrogen concentration provided to stoc k plants on percent survival, visual rooting quality (1=poor; 5=best), tota l dry weight (shoot weight + root weight), and root : shoot ratio of s ub-terminal bougainvillea stem cuttings planted during the cool season (Expt. 2). Source df Percent survival Rooting quality Total dry weight Root : shoot ratio Rep (R ) 5 NS NS NS NS Harvest date (H) 2 0.0001 0.0001 0.0001 0.0001 Evaluation date (W) 1 NS 0.0001 0.0083 0.0001 H x W 2 0.0038 0.0032 0.0001 0.0001 Cultivar (C ) 1 0.0001 0.0732 0.0024 NS H x C 2 NS 0.0001 0.0001 0.0032 W x C 1 NS NS NS NS H x W x C 2 NS NS NS NS N Concn (T) 2 NS NS NS NS H x T 4 NS NS NS NS W x T 2 NS NS NS NS H x W x T 4 0.0480 NS NS NS C x T 2 NS NS NS NS H x C x T 4 NS 0.0024 NS NS W x C x T 2 NS NS NS NS H x W x C x T 4 NS NS NS NS NSNonsignificant at P 0.05.

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50 Table 2-18. Harvest date (H) by cultivar by nitrogen (N) concentr ation interaction for visual rooting quality (1-5; 1=poor, 5= best) of sub-terminal bougainvillea stem cuttings planted during the cool season (Expt. 2). Rooting quality N concn Raspberry Ice Purple Small Leaf (mgL-1) H2 H4 H6 H2 H4 H6 100 3.6 1.8 3 3.2 2.4 2.4 200 3.9 1.7 3 2.6 2.3 3 300 4.1 1.6 2.7 2.5 2.5 3.1 Significance NS NS NS NS NS LSD 0.6 NS, Nonsignificant or significant at P 0.05, respectively. Table 2-19. Influence of harvest date (H), cultivar, and nitrogen (N) concentration on quantity of primary roots of sub-term inal bougainvillea stem cuttings planted during the cool season (Expt. 2). Number of primary roots N concn Raspberry Ice Purple Small Leaf (mgL-1) H2 H4 H6 H2 H4 H6 100 8.8 0.5 3.2 3.3 3.1 2.1 200 10.2 0.5 6.2 2.4 4.6 3.6 300 6.2 1 2.7 2.6 1.7 5.9 Significance NS NS NS NS LSD 2.6 2.8 Source of variance df Pr > F Rep (R ) 5 NS Harvest date (H) 2 0.0001 Cultivar (C ) 1 0.0259 H x C 2 0.0001 N Concn (T) 2 0.0483 H x T 4 NS C x T 2 NS H x C x T 4 0.0059 NS, Nonsignificant or significant at P 0.05.

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51 Table 2-20. Significance levels for effects of harvest date, cultivar, and nitrogen (N) concentration on foliar concentration of macronutrients N, phosphorous (P), potassium (K), calcium (Ca), and magnesium (Mg) of sub-terminal bougainvillea stem cuttings harvested during the cool season (Expt. 2). Source df N P K Ca Mg Rep 2 NS NS NS NS NS Harvest date (H) 2 0.0006 0.0001 0.0001 0.0001 0.0001 Cultivar (C ) 1 0.0213 0.0048 0.0041 0.0001 0.0001 H x C 2 NS 0.0001 0.0003 NS 0.0036 N concn (N) 2 0.0001 NS 0.0001 NS 0.0013 H x T 4 0.0001 NS NS NS NS C x T 2 NS NS NS 0.0024 0.0052 H x C x T 4 NS NS NS 0.0046 0.0330 NSNonsignificant at P 0.05. Table 2-21. Harvest date (H) by cultivar by nitrogen (N) concentr ation interaction for foliar nutrient concentrati on of calcium (Ca) and magnesium (Mg) in subterminal bougainvillea cuttings harvested from stock plants fertigated with N at concentrations of 100, 200, or 300 mgL-1 during the cool season (Expt. 2). Ca (%) N concn Raspberry Ice Purple Small Leaf (mgL-1) H1 H3 H5 H1 H3 H5 100 1.40 1.04 a1.15 1.64 b1.25 1.03 200 1.44 0.79 b0.57 1.70 a1.28 1.38 300 1.56 0.55 c0.76 1.73 a1.25 1.01 NS ** NS ** NS NS Mg (%) N concn Raspberry Ice Purple Small Leaf (mgL-1) H1 H3 H5 H1 H3 H5 100 1.07 0.73 a0.75 1.03 0.81 0.70 200 1.07 0.54 b0.55 1.07 0.76 0.77 300 1.02 0.43 c0.45 1.05 0.73 0.62 NS ** NS NS NS NS NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. z Any two means within a column not followe d by the same letter are statistically different using LSD at P 0.05.

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52 0 1 2 3 4 5 6 7 024681012141618202224 Weeks after pottingpH 100 200 300 Harvest date Figure 2-1. Nitrogen (N) concentration x sa mpling date interacti on for leachate pH collected from bougainvillea stock plants fertilized with N at 100, 200, or 300 mgL-1 during the warm season (Expt. 1). E ach data point represents mean of three samples. Cutting harvests are indicated with a tr iangle along x-axis. 0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 024681012141618202224 Weeks after plantingEC (mS/cm) EC Harvest date Figure 2-2. Main effects for leachate electrical conductivity (EC) over time, as recorded from bougainvillea stock plants during th e warm season (Expt. 1). Each data point represents the mean of three samp les. Cutting harvests are indicated by a triangle along the x-axis.

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53 0 100 200 300 400 500 600 700 800 900 024681012141618202224 Weeks after plantingNO3-N (mgL-1) Raspberry Ice Purple Small Leaf Harvest date Figure 2-3. Nitrogen (N) concentration x samp ling date interaction for nitrate nitrogen (NO3-N) values recorded from stock pl ant leachate of 'Raspberry Ice' and 'Purple Small Leaf' bougainvilleas (Expt. 1) Each data point represents the mean of three samples. Cutting harves ts are indicated by a triangle along the x-axis. 0 1 2 3 4 5 6 7 0246810121416182022 Weeks after plantingpH 100 200 300 Harvest date Figure 2-4. Nitrogen (N) concentration by sa mpling date interact ion for leachate pH collected from bougainvillea stock plants fertilized with N at 100, 200 or 300 mgL-1 during the cool season (Expt. 2). E ach data point represents the mean of three samples. Cutting harvest da tes are indicated by a triangle along the xaxis.

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54 0 0.5 1 1.5 2 2.5 3 3.5 0246810121416182022 Weeks after plantingEC (mS/cm) EC Harvest date Figure 2-5. Main effects for leachate electrical conductivity (EC) over time as recorded from bougainvillea stock plants during th e cool season (Expt. 2). Each data point represents the mean of three samp les. Cutting harvests are indicated by a triangle along the x-axis. 0 200 400 600 800 1000 1200 1400 1600 1800 0246810121416182022 Weeks after plantingNO3-N (mg.L-1) 100 200 300 Harvest date Figure 2-6. Nitrogen (N) concentration by samp ling date interaction for leachate nitrate nitrogen (NO3-N) values recorded from stock plants of bougainvillea fertilized with N at 100, 200 or 300 mgL-1 during the cool season (Expt. 2). Each data point represents the mean of three samp les. Cutting harvests are indicated by a triangle along the x-axis.

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55 CHAPTER 3 LATERAL BRANCHING AGENTS Introduction With the ability to withstand high humidity levels and extremely warm temperatures in southern climates and the versa tility to be used as annuals in the northern landscapes, tropical perennials offering vibr ant color choices and interesting foliar textures for U.S. consumers (Palmer, 2000). Oleander ( Nerium oleander L.) ‘Dwarf Salmon’ and Tecoma [ Tecoma stans (L.) Juss.] ‘Esperanza’ are two highly desirable tropical perennials because of their outstand ing flowering characteristics and superior garden performance. Oleander is a multipurpose tropical pere nnial, hardy to USDA Hardiness Zone 9, that can be used as a foundation plant, hedge or as a featured specimen in a container garden. Once established, oleander is able to withstand poor soil conditions and is reportedly salt tolerant (Walker and Hanl y, 1996). It requires a sunny location for northern gardeners, who could use it as a pot ted plant to be take n indoors during winter months (McCulla, 1973). Tecoma, characteri zed by its trumpet-shaped yellow flowers was recognized as a 2005 “Plant of the Year” by the Florida Nursery, Growers and Landscape Association (FNGLA, 2005). Used as a deciduous shrub or tree and hardy to USDA Hardiness Zone 10, tecoma grows to 7.6 m (25 ft) at maturity, and is equally attractive as a containerized specimen plant. While demand is increasing for both sp ecies, shortages in rooted cutting availability exist amongst shipments to grower s. This, coupled with low quality rooted

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56 liners supplied by domestic propagation firms ha s become a concern for growers wishing to produce oleander and tecoma for sale. As a means of increasing cutting number the effects of ethephon (Florel) [3.9% (2-chlor oethyl) phosphonic acid] (Monteray Lawn and Garden Products, Inc., Fresno, Calif.) and benzyladenine (BA) on stock plant cutting quantity has been proven benefi cial for promotion of axillary shoot development in some floriculture crops such as: chrysanthemum ( Chrysanthemum morifolium ); geranium ( Pelargonium x hortorum L'Her.); poinsettia ( Euphorbia pulcharrima Willd. ex Klotsch); scaevola ( Scaevola aemula R. Br ); and vinca vine ( Vinca major L.) (Gibson and Whipker, 2004; Konjoian, 1994; Witaszek and Mynett, 1989). Because oleander and tecoma display limited branching in containe rs, they are ideal candidates for lateral branching chemical sprays to increase cutting quantity and quality. The objective of the experiment was to determine the efficacy of lateral branching treatments on stock plants of Nerium oleander ‘Dwarf Salmon’ and Tecoma stans ‘Esperanza’and to determine the most effective concentrations of Fres co (N-[phenylmethyl]-1H-purine 6-amine +gibberellins A4A7: 1.8%)(Fine Agrochemicals Ltd., Worchester, United Kingdom), FAL-457 (N-[phenylmethyl]-1H-purine 6-amin e: 2.0%)(Fine Agrochemicals Ltd.), and Florel for increasing harvest quantity, quality, and subseque nt rooting performance of cuttings. Materials and Methods Experiment 1 Rooted liners of tecoma ( Tecoma stans ‘Esperanza’) and oleander ( Nerium oleander ‘Dwarf Salmon’) were planted, 2-per pot, into 16d x 18.5h cm round plastic containers on 18 May 2005. The root substr ate used was Fafard 4-P (Fafard, Anderson, S.C.), which contained: 4 sphagnum peat: 2 pi ne bark: 2 vermiculite : 1 perlite (v/v).

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57 Slow release fertilizer (19N-2.6P-9.9K) was incorporated into the stock plant rooting substrate at a rate of 3.21 gm-3 (1.33 oz/ft3) prior to planting. Plants were then fertigated once-weekly using 14N-1.7P11.6K with N at 200 mgL-1. Plants were established for 4 weeks, until roots reached the bottom of the pot, and then given a hard pinch on 6 July. Stock plants were sprayed, using a volume of 204 mLm3, with a treatment solution consisting of Fresco or FAL457 at 125, 250, or 500 mgL-1 or Florel at 250, 500, or 1000 mgL-1. All treatments were mixed with water containing 0.66 mgL-1 polyether modified polysiloxane (Capsil) (The Scotts Company, Ma rysville, Ohio) spray adjuvant. A control treatment was included, which was sprayed w ith water and adjuvant only. Treatments were applied 7 July, 22 July, 4 Aug., and 24 Aug., and cuttings were harvested 4 and 8 weeks after treatments were initiated, on 3 Aug. and 30 Aug., respectively. Stock plants were assigned to a randomized complete block design (RCBD) with 10 treatments and five single pot replications of each treatment. Experiment 2 The second experiment was repeated in th e same manner as the first, with the following exceptions: Stock plants were transp lanted on 5 Oct. and given a hard pinch on 31 Oct. Lateral branching agents with nine single pot replications of the 10 treatments were applied 1 Nov., 15 Nov., 29 Nov., and 13 D ec. Cuttings were harvested on 28 Nov. and 28 Dec. 2005. Harvesting procedure At each harvest, all 2-node tip cuttings were removed from stock plants of oleander and tecoma. Cutting quantity was recorded al ong with stem length and diameter (cm) of three random cuttings per plant. Cuttings we re then dipped (0.5 cm for 3 s) in a 1,500 mg.L-1 solution containing indolebutyric acid (IBA) 1%: napthaleneacetic acid (NAA)

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58 0.5% and then inserted in a substrate containi ng 4 perlite: 1 vermiculite (v/v). Cuttings were assigned to a completely randomized design (CRD) and propagated under mist (5 s every 20 min). Each species was randomi zed separately and was assigned, when numbers were available, with 6 sub-samp les and 4 replications of each treatment. Cutting evaluations Cuttings were evaluated 5 weeks after pl anting for percent su rvival and rooting quality. Root quality rankings were assigned on a 1 to 5 scale, relative to each species, with a general ranking system as follows: 1=0 roots with some callus formation; 2= minimal rooting, 1 or more primary roots around cutting base; 3=5 or more primary roots with some elongation; 4=uniform root growth and development around cutting base with primary root elongation and some lateral r oot growth; 5= best rooting performance uniform primary and lateral root devel opment with well defined root-ball. All data were subject to analysis of variance using gene ralized linear model procedure in SAS (SAS institute, Cary, N.C.) and means separated using LSD with P 0.05. Results Experiment 1 Harvest yield, cutting le ngth, and stem diameter The species x treatment interaction wa s significant for quantity and length of cuttings, while the harvest date x species x treatment interaction was only significant for stem diameter (Table 3-1). The harvest date x treatment interacti on was significant for cutting length. The main effects of harvest date will be presented separately for cutting quantity.

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59 Table 3-1. Significance levels for effects of harvest date, cultivar, and branching treatment on quantity, length, and stem diameter of 2-node stem cuttings harvested from stock plants of oleande r and tecoma during the cool season (Expt. 1). Source df Quantity Length Diameter Rep 4 NS NS NS Harvest date (H) 1 0.0020 0.0001 0.0026 Cultivar (C ) 1 0.0001 0.0001 0.0001 H x C 1 NS NS NS Branching treatment (T) 9 0.0001 0.0001 0.0001 H x T 9 NS 0.0155 0.0059 C x T 9 0.0001 0.0404 0.0001 H x C x T 9 NS NS 0.0355 NS Nonsignificant at P 0.05. Table 3-2. Cultivar by treatment interacti on for quantity and length of 2-node stem cuttings harvested from stock plants of Tecoma stans and Nerium oleander during the warm season (Expt. 1). Tecoma Oleander Treatment Concn (mgL-1) Quantity Length (cm) Quantity Length (cm) Control 0 12.1 6.5 13.9 4.7 Fresco 125 9.4 7.4 6.8 5.3 Fresco 250 8.8 6.0 3.1 4.9 Fresco 500 3.3 5.4 1.2 2.5 FAL-457 125 10.2 7.1 14.5 3.8 FAL-457 250 11.1 7.0 14.0 3.7 FAL-457 500 9.6 6.3 14.6 4.0 Florel 250 7.9 6.4 14.7 4.3 Florel 500 5.8 5.4 19.0 4.0 Florel 1000 6.3 4.5 17.7 3.5 Significance ** ** ** NS LSD 2.7 1.2 3.3 NS, *, *** Nonsignificant or significant at P 0.05, or 0.001, respectively.

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60 Amongst all treatments and between species the first harvest (9.4) produced 15% less cuttings than the se cond harvest (11.0) (LSD=0.2, n=100). For both species, differences in cutting length were not signi ficant amongst treatments for the first harvest (5.4 cm). During the second harvest, cutti ngs harvested from the control had similar lengths to those harvested from all treatments, except from Fresco at 500 mgL-1, which were 57% shorter (data not s hown). Stock plants treated with Fresco at 125 or 250 mgL1 produced cuttings that were 5.6 and 4.7 cm respectively, which were at least 121% longer than those treated with Fresco at 500 mgL-1 (2.1 cm). Stock pl ants treated with Florel at 250 mgL-1 (5.0 cm) produced cuttings that were 152% longer than Florel at 1000 mgL-1 (3.3 cm); both concentrations we re similar to Florel at 500 mgL-1 (3.8 cm) (LSD=1.64, n=10). Stock plants treated with all concentrations of FAL-457 produced similar sized cuttings (data not shown). At least 129% more tecoma cuttings were harvested from the control than from stock plants treated with Fresco or Florel (Table 3-2). St ock plants treated with FAL-457 produced a similar amount of cuttings to the control. Fresco at 125 mgL-1 produced at least 63% fewer cuttings than at the higher concentrati ons, although cutti ngs harvested from Fresco at 250 or 500 mgL-1 were mostly defoliated with necrotic lesions on stems. Cutting quantity was similar from all Florel treated stock plants. Cutting length was similar to the control for all tr eatments except Florel at 1000 mgL-1 (Table 3-2). At least 122% longer cuttings were harvested from st ock plants treated with Fresco at 125 mgL-1 than at 250 or 500 mgL-1. Stock plants treated with Florel at 250 mgL-1 produced 143% longer cuttings th an at 1000 mgL-1; however both low and high concentrations were

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61 similar to Florel at 500 mgL-1. Cuttings harvested from FAL-457 treated stock plants at any concentration were similar in length. Differences in tecoma stem diameter were not significant amongst treatments for the first harvest (0.28 cm). During the second harvest, cuttings from stock plants treated with Fresco at 125 (0.33 cm) or 250 mgL-1 (0.35 cm) were at least 138% larger in diameter than from stock plants treated with Fresco at 500 mgL-1 (0.24 cm), FAL-457 at 500 mgL-1 (0.24 cm), or Florel at 250 to 1000 mgL-1 (0.20 and 0.22 cm) (LSD=0.9, n=5). All treatments produced si milar sized cutting diameters to the control, except stock plants treated with all concentrations of Fl orel, which were smaller (data not shown). Florel treatments at all concentrations we re similar to each other, as were FAL-457 treatments. Stock plants of oleander trea ted with Florel at 250 mgL-1 or any concentration of FAL-457 produced a similar number of cuttings to the control (Table 3-2). Fresco at 125 mgL-1 produced 219% and 258% more cu ttings than at 250 or 500 mgL-1, respectively, however cuttings were mostly defoliated and exhibited excessive stem elongation with signs of necrosis. Florel at 250 mgL-1 produced 23% fewer cuttings than at 500 mgL-1, however both were similar to Florel at 1000 mgL-1. Differences in oleander cutting length we re not significant amongst treatments (3.8 cm). Cutting stem diameter was similar for all treatments during the first harvest, the same trend occurred in the second harvest, ex cept for smaller stem diameters with Fresco (data not shown). Cutting stem diameter was not measured from stock plants treated with Fresco at 500 mgL-1, as zero cuttings were produced at the second harvest. Cuttings

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62 harvested from stock plants tr eated with Fresco at 250 mgL-1 (0.11 cm) had a 33% smaller stem diameter than at 125 mgL-1 (0.16 cm) (LSD=0.05, n=5). Percent survival and visual rooting quality The harvest date x species x treatment interaction was significant for percent survival and rooting quality (Table 3-3). Differences in percent survival were not significant amongst treatments for the first harvest of tecoma, which exhibited 94% survival (Table 3-4). Rooting quality of cuttings harvested from the control were similar to all treatments except Fresco concentrations of 125 and 250 mgL-1. Fresco at lower concentrations had reduced rooting quality wh en compared to all treatments except when provided at 500 mgL-1. For the second harvest of tecoma, cuttings planted from stock plants treated with Fresco at 500 mgL-1 had at least a 57% lower ra te of survival than all other lateral branching treatment s, which performed similarly to the control (Table 3-4). Rooting quality was similar to the control for cu ttings harvested from stock plants treated with FAL-457 at 125 or 250 mgL-1 or Florel at 250 or 500 mgL-1. Fresco at 125 or 250 mgL-1 had greater root quality ratings than at 500 mgL-1, which was the lowest of any treatment. Stock plants treated with Florel at 250 or 500 mgL-1 had better rooting quality than 1000 mgL-1. Cuttings had better rooting qualit y when planted from stock plants treated with FAL-457 at 250 mgL-1, when compared to 125 or 500 mgL-1. For the first harvest of olea nder, propagules from stock plants treated with Fresco at 250 or 500 mgL-1, or FAL-457 at 125, 250, or 500 mgL-1 exhibited stem and foliar disease symptoms, and did not survive propaga tion. While cuttings harvested from stock plants treated with Florel at all concentr ations performed similarly, treatments of 1000 mgL-1 had a 202% higher survival rate than Fresco treatments at 125 mgL-1 (Table 3-4). Rooting quality ratings were higher for cuttings planted from stock plants treated with

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63 Florel at 1000 mgL-1 than Florel at 500 mgL-1 or Fresco at 125 mgL-1, while Florel at 250 mgL-1 was similar to all three. For the s econd harvest, no cutt ings were planted from stock plants treated with Fresco at 500 mgL-1. As concentrations of Fresco increased from 0 to 250 mgL-1, percent survival and subsequent rooting quality decreased (Table 3-4). Cutti ngs planted from stock plants treated with Fresco at 250 mgL-1 had the lowest rate of survival, when compared to all other treatments. All treatments except the two higher concentrations of Fresco had similar survival to the control. For rooting quality, all treatments we re similar to the control, except Fresco at 125 and 250 mgL-1. Table 3-3. Significance levels for effects of harvest date, cu ltivar, and lateral branching treatment on percent survival, visual r ooting quality (1=poor ; 5=best), total cutting dry weight (root weight + shoot weight) and root : shoot ratio (root weight / shoot weight) of cuttings plan ted from stock plants of oleander and tecoma during the warm season (Expt. 1). Source df Percent survival Rooting quality Total dry weight Root : shoot ratio Rep 3 NS NS NS NS Harvest date (H) 1 0.0001 0.0007 NS 0.0001 Cultivar (C ) 1 0.0001 0.0001 NS 0.0348 H x C 1 0.0001 0.0001 NS 0.0103 Branching treatment (T) 9 0.0001 0.0001 NS NS H x T 9 0.0001 0.0001 NS NS C x T 9 0.0001 0.0001 NS NS H x C x T 9 0.0124 0.0077 NS NS NS Nonsignificant at P 0.05. Cutting total dry weight and root : shoot ratio The harvest date x species interaction was significant for cutting root : shoot ratio (Table 3-3), however differences in total cu tting dry weight were not significant amongst treatments (0.579 g). For the first harvest, differences in root : shoot ratio between species were not significant (0.38). For the second harvest however, tecoma (0.29) had a 242% larger root : shoot ratio than oleander (0.12) (LSD=0.07, n=30).

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64Table 3-4. Harvest date by cultivar by bran ching treatment interaction for rooting qu ality (1=poor; 5=best ) and percent surviv al of 2node stem cuttings propagated from stock plants of Nerium oleander ‘Dwarf Salmon’ and Tecoma stans ‘Esperanza’ harvested on 3 Aug. (H1) and 30 Aug. 2005 (H 2), during the warm season (Expt. 1). Tecoma Oleander H1 H2 H1 H2 Treatment Concn (mgL-1) Survival (%) Rooting quality Survival (%) Rooting quality Survival (%) Rooting quality Survival (%) Rooting quality Control 0 78 4.1 100 4.1 0 0.0 91 3.6 Fresco 125 89 4.2 88 3.4 15 2.0 88 2.8 Fresco 250 100 3.3 96 3.4 0 0.0 13 0.8 Fresco 500 100 3.4 38 2.6 0 0.0 0 0.0 FAL-457 125 100 3.9 100 3.8 0 0.0 96 3.3 FAL-457 250 94 4.1 100 3.2 0 0.0 92 3.4 FAL-457 500 89 4.2 100 4.1 0 0.0 92 2.9 Florel 250 94 4.2 100 4.3 25 3.5 88 3.4 Florel 500 100 4.3 100 4.1 21 2.1 92 3.7 Florel 1000 100 4.1 100 3.3 30 4.4 95 3.4 Significance NS *** *** *** *** LSD 0.6 15 0.5 15 1.8 19 0.8 NS, *, ** Nonsignificant or significant at P 0.05 or 0.001, respectively.

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65 Experiment 2 Harvest yield, cutting le ngth, and stem diameter There was a harvest date x species x tr eatment interaction for all measured parameters except stem diameter (Table 35). The harvest x species and harvest x treatment interactions were both significant for stem diameter. Table 3-5. Significance levels for effects of harvest date, cultivar, and branching treatment on quantity, length, and stem di ameter of cuttings harvested from stock plants of oleander and tecoma during the cool season (Expt. 1). Source df Quantity Length Diameter Rep 8 0.0001 0.0316 NS Harvest date (H) 1 0.0001 NS NS Cultivar (C ) 1 0.0001 0.0198 NS H x C 1 0.0001 0.0001 0.0021 Branching treatment (T) 9 0.0003 0.0001 0.0095 H x T 9 0.0417 NS 0.0108 C x T 9 0.0001 0.0001 NS H x C x T 9 0.0001 0.0020 NS NS Nonsignificant at P 0.05. For the first harvest of tecoma, stock plants treated with FAL-457 at 125 to 250 mgL-1 or Fresco at 125 mgL-1 produced a similar number of cuttings to the control (Table 3-6). Untreated plants produced at least 143% more cuttings than stock plants treated with Fresco at 250 or 500 mgL-1, FAL-457 at 500 mgL-1, or Florel at any concentration. Florel at 250 mgL-1 produced 520% more cu ttings than at 1000 mgL-1, but was similar to 500 mgL-1. Similarly, Fresco at 125 mgL-1 produced 178% more cuttings than 500 mgL-1, yet quantity was similar to 250 mgL-1. Tecoma stock plants treated with Florel at 1000 mgL-1 produced the smallest number of cuttings (Table 3-6) All other treatmen ts performed similarly to the control, except Fresco at 125 and 500 mgL-1, which yielded at least 131% more cuttings. For the second harvest, cutting quantity was similar to the control for all treatments except Florel

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66 at 1000 mgL-1 (Table 3-7). Within chemical, cutting yield was similar, however upon comparison of lateral branching agents at l east a 181% greater numbe r of cuttings were harvested from stock plants treated with FAL-457 at 125 or 250 mgL-1 than from those treated with all concentrations of Fresco or Florel at 1000 mgL-1. Cutting quantity harvested from stock plants treated with all c oncentrations of Florel was similar to all concentrations of Fresco. Table 3-6. Influence of lateral branching ag ents on number and le ngth of cuttings and subsequent rooting performance values for cuttings harvested on from stock plants of Tecoma stans ‘Esperanza’ on 28 Nov. 2005. Treatment Concn (mgL-1) Quantity Length (cm) Rooting quality Total dry weight (g) Root : shoot ratio Control 0 7.0 4.9 4.0 0.488 0.24 Fresco 125 6.3 7.7 3.8 0.460 0.38 Fresco 250 4.9 6.3 3.1 0.296 0.29 Fresco 500 3.6 6.5 2.8 0.219 0.25 FAL-457 125 7.0 5.3 3.5 0.478 0.24 FAL-457 250 5.9 5.3 2.8 0.367 0.22 FAL-457 500 3.8 5.3 3.2 0.386 0.18 Florel 250 2.9 5.5 4.0 0.338 0.30 Florel 500 1.8 4.3 2.8 0.181 0.35 Florel 1000 0.6 1.8 4.2 0.244 0.34 Significance ** ** * LSD 1.9 1.5 0.9 0.142 0.13 *, ** Significant at P 0.05 or 0.01. During the first harvest, cutting length was similar for all treatments as compared to the control, except those from stock plants treated with Fresco at 125 or 500 mgL-1, which had longer cuttings (Table 3-6). Fresco at 125 mgL-1 had longer cuttings than any other treatment, except at c oncentrations of 250 to 500 mgL-1. When comparing individual branching agents, cutting length was similar for all concentrations of Fresco, as were all concentrations of FAL-457. Stock plants treated with concentrations of Florel at 250 to 500 mgL-1 produced at least 241% longer cuttings than at 1000 mgL-1. Cutting

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67 length from the second harvest was similar to th e control for all treatm ents except Fresco at 500 mgL-1 (Table 3-7). Cuttings harvested from stock plants treated with Fresco at 500 mgL-1 were 372% longer than from Florel at 1000 mgL-1. When considering each branching agent individually, cutting length was similar for all concentrations of each chemical applied. Table 3-7. Influence of lateral branching agents on quantity and length of cuttings and subsequent dry weights of cuttings harvested on from stock plants of Tecoma stans ‘Esperanza’ on 28 Dec. 2005. Treatment Concn (mgL-1) Quantity Length (cm) Total dry weight (g) Root : shoot ratio Control 0 3.4 2.5 0.110 0.55 Fresco 125 2.3 4.0 0.210 0.30 Fresco 250 1.8 3.0 0.113 0.39 Fresco 500 2.1 4.9 0.140 0.28 FAL-457 125 4.2 3.4 0.271 0.15 FAL-457 250 4.2 3.7 0.266 0.15 FAL-457 500 3.3 3.5 0.211 0.26 Florel 250 2.6 2.6 0.055 0.50 Florel 500 2.6 2.4 0.037 0.99 Florel 1000 1.2 1.3 0.015 1.64 Significance * ** LSD 1.8 1.9 0.079 0.71 *, ** Significant at P 0.05 or 0.001. At the first harvest of oleander, cutting quantity was similar for stock plants treated with Fresco at 125 mgL-1, Florel at 1000 mgL-1, or FAL-457 at all concentrations (Table 3-8). Additionally, stock plants treated with Fresco at 125 to 250 mgL-1 or Florel at 250 to 500 mgL-1 generated at least 308% more cuttings than untreated stock plants. Results varied in response to con centration for Florel and Fr esco, while FAL-457 at all concentrations performed simila rly. Florel at 250 to 500 mgL-1 produced more cuttings than at 1000 mgL-1, while Fresco at 125 mgL-1 produced fewer cuttings than at

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68 concentrations of 250 to 500 mgL-1. During the second harvest, with 9.4 cuttings per plant, differences in quantity were not significant amongst treatments. Oleander cutting length during th e first harvest was similar for all branching agents as compared to the control, except Fresco at 250 and 500 mgL-1 (Table 3-8). Fresco treatments of 125 mgL-1 produced 38% shorter cuttings than 500 mgL-1, while both concentrations were similar to 250 mgL-1. Cutting length in response to all concentrations of Florel and FAL-457 were si milar. During the second harvest, lengths of cuttings from untreated plants were simila r to FAL-457 at all conc entrations and Florel at 500 mgL-1 (Table 3-9). All concentrations of Florel produced shorter cuttings than cuttings from Fresco treated stock plants Longer cuttings with Fresco at 250 mgL-1 were recorded when compared to 500 mgL-1. When considering both species, at the fi rst harvest and amongst treatments, cutting diameter was 126% larger for tecoma (0.23 cm) than oleander (0.19 cm) (LSD=0.04, n=90). At the second harvest however, olea nder (0.20 cm) had a 118% larger diameter than tecoma (0.17 cm) (LSD=0.02, n=90). Duri ng the first harvest of both species, all branching agents provided a si milar diameter to the contro l (0.18 cm), except Florel at 250 mgL-1 (0.33 cm) which was at least 145% larg er than any other treatment (LSD=0.1, n=18). Stock plants treated with Fresco at 500 mgL-1 (0.23 cm) produced a 183% larger diameter than those treated with Florel at 1000 mgL-1 (0.12 cm), although both were similar to the control. At th e second harvest, most treatments were similar to the control (0.21 cm) except Fresco at 125 mgL-1 (0.16 cm) or 250 mgL-1 (0.15 cm) or Florel at 250 or 1000 mgL-1 (0.17 and 0.13 cm, respectively) (LSD =0.05, n=18). Stock plants treated with FAL-457 at 125 mgL-1 (0.24 cm) or 500 mgL-1 (0.22 cm) produced cuttings with at

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69 least 125% greater diameter th an from those treated with Fresco or Florel at any concentration (data not shown). Relative di fferences amongst concentrations for each chemical were not significant. Percent survival and visual rooting quality There was a harvest date x cultivar x treat ment interaction for both percent survival and visual rooting quality (Table 3-10). Differences in percent survival were not significant amongst treatments for cuttings plan ted from either harvest of tecoma, which were 100% and 77% from harvests 1 and 2, re spectively. Rooting quality from the first harvest of untreated cuttings was similar to most treatments except concentrations of Fresco at 250 to 500 mgL-1, FAL-457 at 250 mgL-1, and Florel at 500 mgL-1, which had lower quality ratings (Table 3-6). Florel treatments at 250 and 1000 mgL-1, were similar, yet produced higher quality ratings than at 500 mgL-1. Although Fresco at 125 mgL-1 had lower rooting quality than at 500 mgL-1, both were similar in quality to Fresco at 250 mgL-1. All concentrations of FAL-457 performed similarly to each other. Florel at 1000 mgL-1 had higher root quality ratin gs than cuttings from stock plants treated with Fresco or FAL-457 at 250 or 500 mgL-1, respectively. Differences in rooting quality for tecoma cuttings from the second harvest were not significant amongst treatments (2.2). Branching agents did not improve percent survival for oleander cuttings planted from the first harvest as compared to the cont rol (Table 3-8). Cuttings planted from stock plants treated with FAL-457 at 250 mgL-1, or Florel at 250 to 500 mgL-1, survived better than those treated with Fresco at 500 mgL-1, FAL-457 at 125 mgL-1, or Florel at 1000 mgL-1. Cuttings had the highest rooting quality va lues from untreated stock plants than from all lateral branching ag ents applied (Table 3-8). FAL-457 treatments provided

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70 similar rooting quality at all concentra tions; however, FAL-457 cuttings had lower rooting quality than Florel treatments at 500 or 1000 mgL-1. Fresco had similar rooting quality values to Florel. Table 3-8. Influence of lateral branching agents on quantity and length, and subsequent rooting for cuttings harvested from stock plants of Nerium oleander ‘Dwarf Salmon’ on 28 Nov. 2005. Treatment Concn (mgL-1) Quantity Length (cm) Survival (%) Rooting quality Total dry weight (g) Root : shoot ratioy Control 0 1.3 2.4 93 4.4 0.522 0.13 Fresco 125 1.9 4.1 93 2.8 0.277 0.27 Fresco 250 4.8 5.4 91 2.8 0.202 0.17 Fresco 500 4.1 6.6 73 2.6 0.112 0.22 FAL-457 125 2.4 2.6 73 2.4 0.317 0.15 FAL-457 250 1.7 1.8 100 1.8 0.293 0.06 FAL-457 500 2.0 2.1 83 1.8 0.281 0.07 Florel 250 5.6 2.1 96 2.6 0.170 0.17 Florel 500 4.2 2.0 100 3.1 0.151 0.22 Florel 1000 1.6 1.6 72 3.2 0.065 0.38 Significance *** *** ** *** LSD 2.0 1.8 21 0.6 0.134 0.11 *, *** Significant at P 0.05 or 0.001. For the second harvest of oleander, cuttings generated from the control had similar percent survival to al l branching treatments except Fres co concentrations of 125 to 500 mgL-1 (Table 3-9). Cuttings planted from stock plants treated with increasing concentrations of Fresco experienced drastic de creases in percent survival. Fresco at 500 mgL-1 had the lowest percent survival. Roo ting quality for cuttings propagated from stock plants treated with FAL-457 at 125 mgL-1 was similar the control, while all other branching treatments were lower in quality (T able 3-9). When considering each chemical individually, Fresco treated cuttings declined in rooting quality as the concentration increased, while concentrations of FAL-457 at 250 to 500 mgL-1 performed similarly,

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71 but with poorer quality as compared to 125 mgL-1. Rooting response to Florel at 250 mgL-1 was similar to 500 mgL-1. Cuttings harvested from stock plants treated with Florel at 500 mgL-1 had higher rooting quality than those treated with Florel at 1000 mgL-1 or Fresco at 250 mgL-1. Cuttings planted from stock pl ants treated with Fresco at 500 mgL-1 had the poorest rooting quality when compared to all other treatments. Table 3-9. Influence of lateral branching ag ents on number and le ngth of cuttings, and subsequent rooting performance values for cuttings harvested on from stock plants of Nerium oleander ‘Dwarf Salmon’ on 28 Dec. 2005. Treatment Concn (mgL-1) Length (cm) Survival (%) Rooting quality Total dry weight (g) Root : shoot ratio Control 3.9 100 4.4 0.539 0.20 Fresco 125 6.7 83 3.0 0.187 0.17 Fresco 250 8.1 37 1.9 0.091 0.17 Fresco 500 6.2 9 1.0 0.108 0.43 FAL-457 125 4.7 100 4.2 0.439 0.20 FAL-457 250 4.2 100 2.8 0.561 0.11 FAL-457 500 3.9 100 3.0 0.590 0.09 Florel 250 3.9 100 3.2 0.217 0.28 Florel 500 2.6 100 3.3 0.181 0.24 Florel 1000 1.9 100 2.5 0.074 0.42 Significance *** ** *** *** *** LSD 1.5 11 0.8 0.092 0.09 *, *** Significant at P 0.05 or 0.001. Cutting total dry weight and root : shoot ratio There was a harvest date x cultivar x tr eatment interaction for total cutting dry weight and root : shoot ratio (Table 3-10). Tecoma cuttings planted from the first harvest were similar in total cutting dry weight to the control when stock plants were provided Fresco at 125 mgL-1 or any concentration of FAL-457 (Table 3-6). Cuttings planted from stock plants treated with Fresco at concentrations of 250 to 500 mgL-1, were at least 36% lower in total dry weight than at 125 mgL-1. Dry weights for all concentrations of FAL-457 were similar, as were all concentrations of Florel. Overall, Fresco or FAL-457

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72 at low concentrations had larger dry weights than Florel at 500 to 1000 mgL-1. Root : shoot ratios for all branching treatments were si milar to the control, except Fresco at 125 mgL-1, which was 161% greater (Table 3-9). Th e root : shoot ratio of Fresco at 125 mgL-1 was greater than at 500 mgL-1 as well as all concentra tions of FAL-457. Fresco and Florel were similar with respect to th e cutting root : shoot ratio, except when FAL457 was applied at 125 mgL-1. For the second harvest, total cutting dry weight of tecoma cuttings propagated from stock plants treated w ith Fresco at 250 mgL-1 or Florel at 250 to 500 mgL-1 were similar to the control (Table 3-7). When consid ering FAL-457, all concentrations had similar dry weights, yet when stock plants were provided concentrati ons of 125 to 250 mgL-1, total cutting dry weights were at least 194% greater than when planted from Fresco treated stock plants at 250 to 500 mgL-1 or all concentrations of Florel. FAL-457 also had increased cutting dry weight as compar ed to the control. Cuttings showed a reduction in total cutting dr y weight when treated with Fresco at 250 to 500 mgL-1 as compared to 125 mgL-1. Fresco at 250 mgL-1 was similar to Florel at 500 to 1000 mgL1. Branching agents applied to tecoma stock plants at all concentrations produced similar root : shoot ratios as compared to the control, except Florel at 1000 mgL-1 (Table 3-7). Root : shoot ratios of Florel at 500 to 1000 mgL-1 were at least 355% higher than Fresco at 500 mgL-1 or any concentration of FAL-457; however, higher concentrations of Florel caused stunting with little to no shoot growth duri ng propagation. Florel at 250 mgL-1 performed similarly to 500 mgL-1 and any concentration of Fresco.

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73 Oleander cuttings from the first harvest ha d at least 165% greater total dry weight with the untreated control than from any other lateral branching agent (Table 3-8). When considering each chemical individually, all c oncentrations of Florel had similar dry weights, as did FAL-457. Fresco ha d greater dry weight at 125 mgL-1 than 500 mgL-1, yet both concentrations were similar to 250 mgL-1. Cuttings planted from stock plants treated with FAL-457 at 250 or 500 mgL-1 had at least 173% greater dry weight than plants treated with Fresco at 500 mgL-1 or Florel at any concentration. Oleander cutting root : shoot ratios were similar to the control for all treatments except for Fresco at 125 mgL-1 or Florel at 1000 mgL-1 which had larger ratios (Table 38). When comparing individual chemicals, all concentrations of Fresco had a similar root : shoot ratio, as did all concentrations of FAL-457. Florel at 250 or 500 mgL-1 had a lower root : shoot ratio than 1000 mgL-1. There was at least a 166% higher root : shoot ratio for cuttings propagated from pl ants treated with Florel at 1000 mgL-1 than from untreated plants, all concentr ations of FAL-457, or Fresco and Florel at 250 to 500 mgL1, respectively. For the second harvest of oleander, cuttings propagated from stock plants treated with FAL-457 at 250 or 500 mgL-1 produced cuttings with a si milar total dry weight to the control (Table 3-9). Cuttings planted fr om untreated stock plants and from those treated with concentrations of FAL-457 at 250 or 500 mgL-1 were at least 123% greater in total cutting dry weight than pl ants treated with FAL-457 at 125 mgL-1. FAL-457 at 250 or 500 mgL-1 were also 248% greater than any c oncentration of Fresco or Florel. Florel had reduced total cu tting dry weight at 1000 mgL-1 as compared to either 250 or 500 mgL-1. Cuttings rooted from Fresco treated stock plants had grea ter dry weight at

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74 125 mgL-1 than at 250 mgL-1, while both of these concentr ations were similar to 500 mgL-1. All treatments had a similar root : shoot ra tio as compared to the control, except FAL-457 at 250 mgL-1, which was smaller, and Fresco and Florel at 500 and 1000 mgL1, respectively, which were both larger. The cu tting root : shoot ra tio was at least 149% greater for cuttings propagated from stock plants treated with Fresco at 500 mgL-1 or Florel at 1000 mgL-1 than from the all other treatments (Table 3-9). Stock plants treated with Florel at 250 mgL-1 had rooted cuttings with at leas t 162% greater r oot : shoot ratio than from plants treated w ith Fresco at 125 to 250 mgL-1, or FAL-457 at 250 to 500 mgL-1. When comparing individual branch ing agents, Fresco at 125 to 250 mgL-1 had a lower root : shoot ratio than 500 mgL-1. Results were similar with Florel concentrations; however, FAL-457 at 125 mgL-1 was greater than 500 mgL-1, while both produced similar results to 250 mgL-1. Discussion During the first experiment, when data wa s pooled between species to investigate the harvest by treatment interaction, both t ecoma and oleander had la rger yield responses at the second harvest as compared to the first, which suggests increased cutting production in response to pinchi ng, rather than chemically influenced. Regardless of treatment, oleander stock plants generated mo re cuttings than tecoma. Lateral branching agents tended to not improve cutting yield of tecoma, rather decreases in yield occurred with Florel and Fresco applications. Oleande r also negatively responded to Fresco at all concentrations, however yield was enhanced when stock plants were treated with Florel. All concentrations of FAL-457 produced similar numbers of cu ttings to untreated plants for both species. The application of a la teral branching agent produced similar sized

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75 oleander cuttings, when compared to the contro l, however slightly shorter cuttings than the control were produced when Florel was provided at increasi ng concentrations to tecoma. Stem diameter was generally not infl uenced by application of a branching agent, although Florel treatments had smaller stem di ameters than untreate d cuttings during the second harvest of both species. Application of branching agents did not in fluence percent surviv al of tecoma from the first harvest, while the s econd harvest produced cuttings with a lower survival rate when Fresco was provided to stock plants as compared to the control. Subsequent rooting quality was also reduced for cuttings planted from Fresco treated stock plants, while all other treatments were similar to the control when planted from both harvests. Phytotoxicity may have increased diseas e incidence, however similar symptoms were observed with the control, which may be result of disease pressure. From the second harvest phytotoxicity of Fresco sprays were evident as percent survival and subsequent rooting quality decr eased as concentrations of Fresco increased, otherwise rooting was similar for all treatments, when compared to the contro l. Dry weight was generally similar for all treatments and for both species, although tecoma cuttings from the second harvest had a larger ro ot : shoot ratio than oleander. During the second study, phytotoxicity in re sponse to treatment applications was similar to the first experiment, although overall performance of cuttings during the second experiment was genera lly improved by comparison. During the first harvest, tecoma showed a reduction in cutting quantity with increasing concentrations of branching agen ts applied, although Fr esco and FAL-457 at lower rates tended to be similar to the contro l. Length of cuttings was excessive when

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76 stock plants were treated with Fresco and stunted when trea ted with Florel, as compared to cuttings harvested from unt reated stock plants. With few exceptions, branching agents applied at higher concentrati ons to stock plants of tecoma caused a reduction in subsequent rooting quality and total cutting dry weight when compared to the control. Root : shoot ratios however, tended to be similar to the control for all treatments, improved ratios occurred when Fresco or Florel was applied at 125 or 500 mgL-1, respectively. A reduction in cutting number occurred for all treatments during the second harvest, when compared to the first. The number of tecoma cuttings was similar for both treated and untreated stock plants, except when treated with Florel at 1000 mgL-1. Cutting length was also shorter during the sec ond harvest, when compared to the first. Supplemental applications of lateral branching agents produ ced similar length cuttings except with Fresco at 500 mgL-1. Stock plants treated with Fresco at 500 mgL-1 produced cuttings with the same length (4.9 cm ) as the untreated stock plants during the first harvest; therefore no cuttings would be considered excessive in length from the second harvest. Benefits to supplemental app lication of branching agents to stock plants were not shown with percent survival and rooting quality because all treatments were similar to the untreated cuttings. Differences in total cut ting dry weight follow a similar trend with differences in cutting length. Cuttings from stock plants treated with Florel at 1000 mgL-1 were shorter than the contro l, but also lower total cut ting dry weight. Phytotoxic responses such as shoot tip desiccation, defolia tion, or excessive stunting as a result of Florel applied at 1000 mgL-1 caused an impractically high ro ot : shoot ratio, otherwise all

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77 branching treatments were similar to the control. In studies conducted on other floriculture species, undesirable stunting has been reported in response to Florel (Faust and Lewis, 2005). During the first harvest of oleander, cu tting quantity was improved when Fresco and Florel were applied at 250 to 500 mgL-1; all other treatments were similar to the control. Compared to tecoma, oleander stoc k plants did not show the same reduction in yield with application of branching agents. In creasing concentrations of Florel applied to stock plants decreased yield, al though quantities harvested were still greater than or equal to the control. Generally lengths of cutti ngs were similar to th e control, yet were excessive when harvested from stock plants trea ted with Fresco at higher concentrations. All treatments rooted with similar percent survival when compared to the control, except for reduced survivability with Florel at 1000 mgL-1. Despite similar percent survival, all treatments had a reduced visual rooting quali ty when compared to the control. Among the least affected by treatmen t with respect to root quality rankings were those harvested from stock plants treated w ith Fresco and lower concentrations of FAL457 or Florel. Both total cutt ing dry weight and the root : shoot ratio seem to coincide with differences observed with rooting quality. During the second harvest, all treatments produced the same number of cuttings, yet comparable to other harvests, cutting le ngth was similar for most treatments with stunting and stretching of propagules when st ock plants were trea ted with Florel or Fresco, respectively. Percent survival was 100% for all trea tments except poor survival rates associated with increasing concentrations of Fresco. Rooting quality was also reduced with all Fresco concentrations and qua lity declined as concentration increased.

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78 Total cutting dry weight was lower than the control for all treatments except FAL-457 at 250 to 500 mgL-1, which were similar. Lower total cutting dry weights, coupled with increased root : shoot ratios for both Fresco at 500 mgL-1 and Florel at 1000 mgL-1 reflected desiccation of the shoot tip or stunting, respectively. Similar root : shoot ratios for nearly all other treatments were similar, which may be a better indication of rooting performance relative to cutting size than visual evaluations, therefore oleander cuttings performed similarly to the control when treate d with branching agents prior to the second harvest. Fresco applications to the stock plants exhibited many phytotoxic symptoms that would be considered highly undesirable for cutting propagation. Similar phytotoxic responses were observed when Fresco was applied to stock plants of Clematis spp. and Rhododendron spp. (Bell et al., 1997; Puglis i, 2002). Symptoms were more severe on oleander than tecoma. The undesirable effects of BA+GA4+7 (active ingredient in Fresco) have been attributed to spray burn as a result of BA in the treatment formulation (Bell et al., 1997). When Clematis spp., were treated with different ratios of BA and GA to reduce phytotoxicity, all combinations had similar effects. The 1:1 ratio (similar formulation to Fresco) produced the lowest degree of phytotoxicity (Puglisi, 2002). Cuttings harvested from stock plants trea ted with FAL-457 did not express the same symptoms; therefore it is hypothesized that the phytotoxicity is a species-dependant outcome, and may be the result of other com ponents such as the chemical carrier. Concentrations of FAL-457 at 125 mgL-1 were shown to improve yield in oleander, but reduced success in propagation limits its use by growers. In tecoma, FAL457 produced similar or lower quality cutti ngs than the untreat ed plants. Although

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79 application of FAL-457 was not shown to be c onsistently beneficial to either species; further investigation should be conducted to determine the efficacy of FAL-457 as a possible tank mix with other formulations. Perhaps FAL-457 and Florel could be mixed to capitalize on the benefits to yiel d and rooting of both treatments. Applications of Florel at 250 to 500 mgL-1 to stock plants of oleander were shown to increase yield with similar rooting when co mpared to the control, yet results were not consistent. Florel applica tions to stock plants of t ecoma were not shown to be advantageous. Variability in response to Flor el for increasing cutti ng quantity has been reported with many floriculture species (Faust and Lewis, 2005), theref ore benefits to its application may been seen at lo wer concentrations or with other tropical species. Further study should be conducted on the efficacy of th ese and other lateral branching agents during other seasons and on other tropical species. Conclusion At the concentrations investigated, none of the lateral branching agents was proven to be consistently advantageous when co mpared to untreated plants. Phytotoxic responses were shown that could potentia lly damage crops in production, therefore growers should consider testing sprays on a sm aller portion of plants before applications are made to the entire greenhouse. The co st of applying these products should be compared to the benefits achieved in producti on. While tecoma and oleander appeared to not benefit from branching treatments at th e concentrations inves tigated, trends were observed that may assist future research.

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80 Table 3-10. Significance levels for effects of harvest date, cu ltivar, and lateral branching treatment on percent survival, visual r ooting quality (1=poor ; 5=best), total cutting dry weight (root weight + shoot weight) and root : shoot ratio (root weight / shoot weight) of cuttings plan ted from stock plants of oleander and tecoma during the cool season (Expt. 2). Source df Percent survival Rooting quality Total dry weight Root : shoot ratio Rep 3 NS NS 0.0106 NS Harvest date (H) 1 0.0001 0.0001 0.0001 0.0004 Cultivar (C ) 1 NS NS NS 0.0001 H x C 1 0.0012 0.0001 0.0001 0.0198 Branching treatment (T) 9 0.0001 0.0001 0.0001 0.0001 H x T 9 0.0001 0.0004 0.0001 0.0257 C x T 9 0.0001 0.0001 0.0001 0.0454 H x C x T 9 0.0009 0.0001 0.0013 0.0278 NS Nonsignificant at P 0.05.

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CHAPTER 4 ROOTING HORMONES Introduction Vegetative propagation has become an es tablished system for reproducing tropical plants. Treating stem cuttings with auxin in creases rooting percentage, accelerates root formation, and enhances uniformity of rooting (Davis et al., 1988). Th is is important in the production of tropical plan ts because many species ofte n require extensive rooting time in propagation where there is an inhere nt risk of air and waterborne pathogens (Howard, 1994). In order to limit time in propagation, studies have been conducted on the stimulatory influence of auxin on numer ous difficult-to-root species (Bhattacharjee and Balakrishna, 1983; Bhat et al., 1988; Czek alski, 1989). Preference for the sythentic compounds indolebutyric acid (IBA) and napt haleneacetic acid (NAA) compared to naturally occurring, indoleacetic acid (IAA), is illustrated by the large number of commercially-available rooting products cont aining one or both in solution (Blazich, 1988a). There are also potassium salt (K+) formulations availa ble that enable IBA (KIBA) and NAA (KNAA) to be dissolved in water, which may be beneficial for some tropical species expressing sens itivity to alcohol-based form ulations. Certain tropical species, such as oleander ( Nerium oleander L.) and bougainvillea ( Bougainvillea glabra Choisy.), have been shown to benefit from appli cations of talc-based formulations at concentrations of 3,000 mg.L-1 IBA and 4,000 to 16,000 mg.L-1 IBA, respectively (Hartmann et al., 2002). For other difficultto-root tropical speci es an evaluation of

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82 commercially-available rooting hormones shoul d be conducted to de termine the optimum concentration to achieve the highest rooting percentage. The objectives of our experiment were to determine the optimum concentration of water-soluble KIBA necessary to effectively ro ot stem cuttings of select tropical species and to compare the viability of KIBA as an alternative rooting hormone to an industry formulation containing IBA 1%: NAA 0.5% at 1500 mgL-1. Materials and Methods Experiment 1 Rooted liners of Allamanda schottii (Pohl.) Bougainvillea glabra (Choisy.) ‘California Gold’ and ‘Helen Johnson’, Mandevilla splendens (Hook. f.) ‘White’, and Nerium oleander (L.) ‘Dwarf Salmon’ were plante d, 3-per pot, into 16 d x 18.5 h cm round plastic containers on 18 April 2005. Th e root substrate used was Fafard 4-P (Fafard, Anderson, S.C.), which contained: 4 sphagnum peat: 2 pine bark: 2 vermiculite: 1 perlite (v/v). Controlled re lease fertilizer (19N-2.6P-9.9K ) was incorporated into the substrate prior to planting, at a rate of 3.21 gm-3 (1.33 oz/ft3). The plants were then provided a once-weekly liquid fertiliza tion of 14N-1.7P-11.6K with N at 200 mgL-1. Plants were established for 4 weeks, until r oots reached the bottom of the pot, and then provided a soft pinch to encourage canopy gr owth on 16 May. Cuttings were harvested every three weeks, on 6 June (H1), 27 J une (H2), 18 July (H3), and 8 Aug. (H4). Mandevilla was harvested three times on differe nt dates with H1, H2, and H3 occurring 6 June, 8 Aug. and 29 Aug., respectively. Phot osynthetic photon flux (PPF) was recorded each day at or near solar noon using a Qu antum Meter (Spectrum Technologies, EastPlainfield, Ill.). Average PPF was 453, 586, and 426 molm-2s-1, for June, July, and Aug., respectively.

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83 Harvesting procedure At each harvest, 6.0 cm terminal stem cuttings were removed from stock plants of allamanda and oleander. Sub-terminal bougainvillea ‘Californi a Gold’ and ‘Helen Johnson’ stem cuttings were harvested in le ngths of 6.5 cm, measured 0.5 cm above first mature leaf. Mandevilla was harvested by removing all 2-node, sub-terminal, stem cuttings 1 cm above and below the top and bottom nodes. Cuttings, with bottom leaves removed, were inserted into perforated plas tic bags with stem bases wrapped in moist paper towels. The bags were placed into boxes, along with a sensor (Hobo Pro, model number H08-032-08) (Onset Computer, Bourne, Mass.) that recorded temperature and relative humidity every 15 s. The boxes were stored ov ernight in a cooler at 10 C. Boxes were sealed and then cuttings were shipped 24 h, to simulate industry ha ndling. Upon arrival, boxed cuttings were left intact until the fo llowing morning, unpacked and treated with rooting hormones before planting, totaling a pproximately 72 h from excision to planting (Table 4-1). Cuttings were dipped 0.5 cm for 3s, in a solution containing 1% indolebutyric acid (IBA) : 0.5% napthaleneacetic acid (NAA) (D ip n’ Grow, Dip n’ Grow Inc., Clackamas, Ore.) at 1,500 mg.L-1 or Indolebutyric acid, pot assium salt (KIBA) at 1,500, 3,000, or 6,000 mgL-1 and then inserted in a s ubstrate containing 4 perlite: 1 vermiculite (v/v). Cuttings were assigned to a completely randomized design (CRD) and propagated under mist (5s every 20 min). E ach species was randomized separately and was assigned, when numbers were available, with 6 sub-samples and 4 replications of each treatment. Sub-sample means were pooled to reduce variability of the replicate. Cutting evaluations Cuttings were evaluated on two occasions from each harvest. Allamanda was evaluated 2 and 4 weeks after planting (WAP), oleander and mandevilla 4 and 6 WAP,

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84 and both bougainvillea cultivars 5 and 7 WAP. At the first evaluation of each harvest percent survival, rooting qualit y, number of primary roots ( 1.0 mm), number of lateral roots, total cutting dry weight (root weight + shoot weight), and cutting root : shoot ratio (root weight/ shoot weight) were recorded. At the second evaluation all other parameters except root number were recorded. Visual ro oting quality ratings were assigned on a 1 to 5 scale, relative to each species, with a general ranking system as follows: 1=callus formation, with no roots; 2=minimal rooti ng; 3=some root growth and development; 4=increased root growth and development; 5= best rooting performance. Cuttings were prepared for dry weight measurement by exci sion of the rooted portion 1 cm above the cutting base, and then placed in a drying oven for 3 d at 70 C. All data were subjected to analysis of variance using ge neralized linear model procedure in SAS (SAS institute, Cary, N.C.) and means separated using LSD with P 0.05. Experiment 2 The second experiment was replicated in the same manner as the first experiment, with the following exceptions: Stock plants were transplant ed 1 Aug., and given a soft pinch on 17 Aug. 2005. Cuttings were again harvested every three weeks on 6 Sept. (H1), 27 Sept. (H2), 18 Oct. (H3), and 8 Nov. (H4). Average PPF was 596, 647, and 355 molm-2s-1, for Sept., Oct., and Nov., respectively. Cuttings were handled in the same manner as the first experiment, except were sh ipped to Gainesville, Fla. using Federal Express 24 h service (Table 4-1).

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85 Results Allamanda schottii Experiment 1 There was a harvest date x evaluation w eek x treatment interaction for percent survival (Table 4-2). Differences in pe rcent survival were not significant amongst treatments for all cutting ev aluations, except those from H3, 2 WAP and H4, 4 WAP. For cuttings planted from H3 and evaluated 2 WAP, all rooting hormone concentrations provided similar percent survival and were greater than the c ontrol, except those treated with KIBA at 1500 mgL-1, which were similar to all treatments. Cuttings evaluated 4 WAP from H4 were all similar to the cont rol when treated with any rooting hormone concentration except Dip n’ Grow at 1500 mgL-1, which had better survival. Concentrations of KIBA at 3000 mgL-1 had similar percent surviv al to Dip n’ Grow at 1500 mgL-1, and expressed greater survivabili ty than KIBA at 1500 or 6000 mgL-1. Generally, cuttings evaluated 4 WAP had a si milar percent survival to 2 WAP, except those treated with KIBA at 6000 mgL-1 and planted from H4, which were greater 2 WAP than 4 WAP. When evaluati ng cuttings 2 WAP, all treatments had a similar response throughout the experiment; some differences occured with respect to harvest date 4 WAP. For untreated cuttings or cu ttings treated with KIBA at 3000 mgL-1, all harvest dates had similar percent surv ival 4 WAP. Cuttings treate d with Dip n’ Grow at 1500 mgL-1 had similar percent survival at all ha rvest dates except H3, which was lower. When cuttings treated with either KIBA at 1500 or 6000 mgL-1 were evaluated 4 WAP, all harvest dates were similar, except H4, which produced lower survival rates. There was a harvest x treatment interacti on for rooting quality (Table 4-3); main effects for evaluation week will be presente d separately. Cuttings evaluated 4 WAP had

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86 higher root quality rankings than at 2 WAP (data not shown). Differences in rooting quality were not significant amongst treatments for H1, H2, or H4. For cuttings planted from H3, rooting hormone applications at any concentration had better rooting quality, when compared to the control, excep t those treated with KIBA at 1500 mgL-1. Rooting quality of cuttings treated with KIBA at 1500 mgL-1 was similar to all treatments except was lower than KIBA at 3000 mgL-1. Cuttings treated with KIBA at 3000 mgL-1 had greater rooting quality than cutti ngs treated with KIBA at 1500 mgL-1 or the control. When considering cuttings that were unt reated or treated with KIBA at 1500 mgL-1, those planted from H4 were similar to all ot her harvests, while cutt ings planted from H1 or H2 had greater rooting quality than H3. Wh en cuttings were treated with Dip n’ Grow at 1500 mgL-1, all harvest dates were similar, excep t H3. Rooting quality was similar for all harvest dates when treated with KIBA at 3000 or 6000 mgL-1, respectively. Table 4-3. Harvest date (H) by treatment in teraction for visual rooting quality (1-5; 1=poor, 5=best) of allamanda cuttings planted during the warm season (Expt. 1). Treatment Concn (mgL1) H1 H2 H3 H4 LSD Control 0 3.9 3.5 2.5 3.2 0.9 Dip n' Grow 1500 4.1 4.3 3.2 4.2 ** 0.5 KIBA 1500 4.2 3.9 3.1 3.7 0.7 KIBA 3000 4.1 4.1 3.9 3.9 NS KIBA 6000 4.3 4.3 3.8 3.8 NS Significance NS NS ** NS LSD 0.7 NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt There was a harvest date x treatment intera ction for number of primary roots. From H1, cuttings treated with KIBA at 1500 to 3000 mgL-1 had a similar number of primary

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87 roots to the control (Table 44). Dip n’ Grow at 1500 mgL-1 had more primary roots than the control and KIBA at 6000 mgL-1 rooted similarly to all treatments except the control. From H2, cuttings treated with KIBA at 6000 mgL-1 had at least 159% more primary roots than the control or cutti ngs treated with KIBA at 1500 or 3000 mgL-1. While Dip n’ Grow at 1500 mgL-1 had a similar number of primary roots to all treatments except the co ntrol, KIBA at 1500 mgL-1 produced a similar number of primary roots to the control and to all treatments except KIBA at 6000 mgL-1. For cuttings planted from H3, the untreated cu ttings produced a similar number of primary roots to both KIBA and Dip n’ Grow at 1500 mgL-1, respectively. Cuttings treated with KIBA at 3000 to 6000 mgL-1 had at least 199% more primary roots than KIBA at 1500 mgL-1 or the control, while cuttings treate d with Dip n’ Grow at 1500 mgL-1 produced a similar number of primary roots to all treatment s. From H4 all treatments were similar to the control except Dip n’ Grow at 1500 mgL-1, which had at least 161% more primary roots than all othe r treatments. When comparing harvest dates, bot h the control and KIBA at 1500 mgL-1 had a greater quantity of primary roots from H1 than other harvest dates (Table 4-4). Similarly, Dip n’ Grow at 1500 mgL-1 had more primary roots when planted from H1 than H3 or H4; however, H2 was similar to all harvest date s. When considering only cuttings treated with KIBA at 6000 mgL-1, H2 had more primary roots pe r cutting than H3, yet both were similar to H1. Cuttings evaluated from H4 pr oduced the lowest number of primary roots as compared to all other harvest dates. Cuttings treated with KIBA at 3000 mgL-1 produced a similar number of primary roots, 18.4 per cutting, amongst harvest dates.

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88 Table 4-4. Harvest date by treatment interac tion for number of primary roots of stem cuttings of allamanda planted during the warm season (Expt 1). Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 17.8 9.2 6.7 7.1 7.4 Dip n' Grow 1500 38.6 25.7 12.9 20.5 14.9 KIBA 1500 24.8 13.4 10.4 10.6 7.6 KIBA 3000 18.7 21.7 20.6 12.7 NS KIBA 6000 30.3 34.5 21.3 11.6 9.7 Significance * * LSD 11.9 12.3 8.5 7.2 NS, Nonsignificant or significant at P 0.05, respectively. Dip n’ Grow = 1% indolebutyric ac id : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt Table 4-5. Harvest date (H) by treatment in teraction for number of lateral roots of allamanda cuttings planted duri ng the warm season (Expt. 1). Number of lateral roots Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 33.6 2.0 0.6 10.2 *** 13.1 Dip n' Grow 1500 18.2 0.8 14.0 31.2 ** 13.8 KIBA 1500 42.9 4.8 5.6 16.5 *** 13.6 KIBA 3000 43.8 3.1 7.6 20.0 *** 7.9 KIBA 6000 27.5 1.4 11.2 21.6 17.5 Significance NS NS NS LSD 8.5 NS,*, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt

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89 There was a harvest date x treatment interaction for number of lateral roots. Differences in quantity of lateral roots were not significant amongst treatments for cuttings evaluated from H1, H2, or H4. Fo r cuttings evaluated from H3, those treated with KIBA at 1500 or 3000 mgL-1 had a similar number of la teral roots to the control (Table 4-5). Dip n’ Grow at 1500 mgL-1 had at least 250% more lateral roots than KIBA at 1500 mgL-1 or the control, yet all concentratio ns of KIBA were similar. When considering only the control, all harvests were similar except H1, which had a greater quantity of lateral roots than the re st. When Dip n’ Grow at 1500 mgL-1 was applied, the number of lateral roots decreased considerably when cuttings were propagated from H2, as compared to H1. For H3 Dip n’ Grow at 1500 mgL-1 produced a similar amount of lateral roots to H1 and H2. For H4 the num ber of lateral roots produced was similar to H1 and greater than H2 or H3. KIBA at 1500 mgL-1 had similar rooting with all harvests except H1, which had the highest numbers of la teral roots, comparatively. When treated with KIBA at 3000 mgL-1, cuttings planted from H2 and H3 were similar and at least 62% and 83% lower than H1 and H4, respectively. Cuttings treated with KIBA at 6000 mgL-1 had the lowest quantity of lateral root s from H4 as compared to other harvest dates. Harvest 2 rooted bette r than H3, yet both had a simila r lateral root number to H1. There was a harvest date x treatment interaction for both cutting total dry weight and root : shoot ratio. There was also a ha rvest date x evaluation week interaction for total cutting dry weight. When cuttings we re planted from H1, all treatments were similar to the control except KIBA at 3000 mgL-1, which was similar in total cutting dry weight to KIBA at 6000 mgL-1 and Dip n’ Grow at 1500 mgL-1. For cuttings planted from H2, H3, or H4, differences in total cu tting dry weight were not significant amongst

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90 treatments. When considering only untreated cuttings, total dry weight was similar for H1 and H4, which were greater than H3; harv est 3 was similar to H2. Additionally, H2 had a similar dry weight to H4 and H3, but was less than H1 (Table 4-6). Cuttings treated with Dip n’ Grow at 1500 mgL-1 had similar total dry weight for H1, H2, and H4, all of which were at leas t 154% greater than H3. Response to KIBA at 1500 mgL-1 amongst harvest dates was similar to the control. Differences in total cutting dry weight were not significant amongst harvest date s when treated with KIBA at 3000 mgL-1 (182 mg). Cuttings treated with KIBA at 6000 mgL-1 had greater total dry weight from H1 than all other harvests. For cuttings planted from H1, t hose evaluated 4 WAP had higher total dry weight than those ev aluated 2 WAP (data not shown). Untreated cuttings planted from H1 had a similar root : shoot ratio to cuttings treated with any concentration of KIBA (Table 4-7). Cuttings treated with Dip n’ Grow at 1500 mgL-1 had a greater root : shoot ratio than most treatments except were similar to KIBA at 6000 mgL-1. For H2, differences in root : shoot ratio were not significant amongst treatments. For cuttings planted fr om H3, all rooting hor mone concentrations showed improved root : shoot ratios when comp ared to the control. When treated with KIBA at 6000 mgL-1, cuttings had the highest root : shoot ratio, and while KIBA at 3000 mgL-1 and Dip n’ Grow at 1500 mgL-1 were similar, both were larger than KIBA at 1500 mgL-1 or the control. When considering only untreated cutt ings, those planted from H4 had similar root : shoot ratio to those planted from all other harvests (Table 4-7). Cuttings planted from H1 had a similar root: shoot ratio to H2 and H4, yet were greater than those planted from H3. Untreated cuttings from H3 were similar to all other harvests except H1. When considering Dip n’ Grow at 1500 mgL-1 and KIBA at

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91 concentrations from 1500 to 6000 mgL-1, differences in root : shoot ratio were not significant amongst harvest date (Table 4-7). Table 4-6. Harvest date by treatment interacti on for total cutting dry weight of allamanda cuttings planted during the warm season (Expt. 1). Total cutting dry weight (mg) Treatment Concn (mgL-1) H1 H2 H3 H4 Significance LSD Control 0 247 142 124 198 67 Dip n' Grow 1500 219 197 128 223 NS KIBA 1500 260 166 136 209 ** 54 KIBA 3000 200 172 159 198 NS KIBA 6000 241 167 139 171 ** 46 Significance NS NS NS LSD 40 NS, *, ** Nonsignificant or Significant at P 0.05 or 0.01, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt Table 4-7. Harvest date by treatment interac tion for the cutting root : shoot ratio of allamanda cuttings planted duri ng the warm season (Expt. 1). Root : shoot ratio Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 0.47 0.34 0.23 0.30 0.17 Dip n' Grow 1500 0.62 0.42 0.52 0.45 NS KIBA 1500 0.47 0.32 0.35 0.37 NS KIBA 3000 0.42 0.36 0.52 0.40 NS KIBA 6000 0.51 0.45 0.60 0.40 NS Significance NS *** NS LSD 0.11 0.06 NS, *, **, *** Non significant or significance at P 0.05, 0.001, or 0.0001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt Experiment 2 There was a harvest date x treatment intera ction for percent survival; main effects for evaluation week are presented separately. Differences in percent survival were not significant amongst treatments for cuttings pl anted from H1, H3, or H4 (98%, 96%, and

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92 93%, respectively). From H2, all treatments had similar percent surv ival (98%), except those treated with KIBA at 3000 mgL-1 (83%) which was lower th an any other treatment (LSD=9, n=7). When considering each treat ment individually, all treatments had similar rate of survival throughout the duration of the second expe riment, except KIBA at 3000 mgL-1 from H2 (data not shown). When co mparing evaluation weeks, all cuttings evaluated 2 WAP had similar survival to those evaluated 4 WAP (96%). There was a harvest date x evaluation week x treatment interaction for rooting quality (Table 4-8). Untr eated cuttings and those tr eated with KIBA at 1500 mgL-1 planted from the first harvest and evaluated 2 WAP had similar rooting quality. Cuttings treated with Dip n’ Grow at 1500 mgL-1 had the best rooting quality when compared to all other treatments, except KIBA at 3000 mgL-1. KIBA at 1500 mgL-1 had reduced quality compared to 3000 mgL-1; however both were similar to 6000 mgL-1. For cuttings evaluated 4 WAP, all treatments were similar to the control, except Dip n’ Grow at 1500 mgL-1, which was also similar to KIBA at concentrations of 3000 to 6000 mgL1. All concentrations of KIBA had similar r ooting quality. For cuttings planted from the second harvest, the control ha d lower rooting quality than any rooting hormone treatment at both 2 and 4 WAP. Overall rooting qual ity was lowest at the third harvest and differences amongst treatments were not signifi cant at either 2 or 4 WAP. For cuttings planted from the fourth harvest, differences amongst treatments were not significant for those evaluated 4 WAP (3.7). At 2 WAP ho wever, all rooting hormone treatments performed similarly to each other and were al l greater than the control, except KIBA at 1500 mgL-1, which was similar. When comparing evaluation weeks for each treatment, cuttings evaluated 4 WAP generally had grea ter root quality ratings than 2 WAP.

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93 Rooting quality was also less affected by hormone treatment or evaluation week as the season progressed, although overall quality at H4 tended to be lower than other harvest dates. The harvest date x treatment interaction was significant for number of primary roots. The number of primary roots was at least 214% greater for cu ttings treated with Dip n’ Grow at 1500 mgL-1 or KIBA at 3000 mgL-1, than for the control or KIBA at 1500 mgL-1 (Table 4-9). Cuttings treated with KIBA at 1500 or 6000 mgL-1 had a similar number of primary roots to the control, while all treatments were similar to KIBA at 6000 mgL-1. Number of primary roots for cutti ngs planted from H2 was highest when treated with KIBA at 6000 mgL-1 and lowest for the control. Dip n’ Grow at 1500 mgL1 and KIBA at 1500 or 3000 mgL-1 had a similar number of prim ary roots. In general, the third harvest had very few primary r oots, 0.2 per cutting, and differences amongst treatments were not significant. For the H4 evaluation, treatments of KIBA at 1500 or 3000 mgL-1 were similar to the control. KIBA at 6000 mgL-1 had at least 192% more primary roots than either th e control or KIBA at 1500 mgL-1, while 3000 mgL-1 was similar to all treatments. Similarly, Dip n’ Grow at 1500 mgL-1 had a similar numbers of primary roots to all treatments, except the control. When considering only untreated cuttings, differences in primary root counts were not significant amongst harvest dates. Those cuttings treated with Dip n’ Grow at 1500 mgL-1 had a greater number of primary roots than from H3; while H2 had more than H1, both H1 and H2 were similar in number to H4. KIBA at 1500 mgL-1 had more primary roots from H4 than H3, while both were simila r to H1. Cuttings planted from H2 had a greater number of primary roots than any other harvest. For cuttings treated with KIBA

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94 at 3000 mgL-1, those planted from H2 had a greater number of primary roots than H4, both being similar to H1, while H3 again, had th e lowest number of any harvest. Cuttings treated with KIBA at 6000 mgL-1 produced a different quantity of primary roots with each harvest evaluation. Cuttings planted from H2 had the largest amount of roots, while H4 produced more than H1. Cuttings planted from H3 produced the fewest number of primary roots. The harvest date x treatment interaction was not significant for number of lateral roots; therefore the main effects will be pres ented. Allamanda cuttings generally had low lateral root numbers, with less than one per cutting and differences were not significant amongst treatments. When comparing harvest da tes, cuttings planted from H4 had more lateral roots than at any othe r harvest (data not shown). Table 4-9. Harvest date by treatment inte raction for number of primary roots of allamanda cuttings planted duri ng the cool season (Expt. 2). Number of primary roots Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 6.5 2.2 0.0 4.7 NS Dip n' Grow 1500 16.1 22.7 0.3 20.8 *** 6.2 KIBA 1500 5.4 19.8 0.0 11.5 *** 6.6 KIBA 3000 13.8 21.2 0.2 12.6 *** 7.5 KIBA 6000 10.8 44.7 0.0 22.1 *** 6.5 Significance ** *** NS LSD 6.0 8.5 9.8 NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, 0.001, respectively. Dip n’ Grow = 1% indolebutyric ac id : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt There was a harvest date x evaluation week interaction for total cutting dry weight. For H1, H2, and H4, cuttings evaluated 4 WAP had a higher total cutt ing dry weight than those evaluated 2 WAP (data not shown). Cuttings planted from H3 had similar weights at both 2 and 4 WAP.

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95 There was a harvest date x evaluation week x treatment interaction for the cutting root : shoot ratio (Table 4-10). For cuttings planted from the first harvest and evaluated 2 WAP, concentrations of KIBA at 1500 or 3000 mgL-1 produced a similar root : shoot ratio to the control with a 31% lower ratio when compared to Dip n’ Grow at 1500 mgL1. KIBA at 6000 mgL-1 produced cuttings with the highest root : shoot ratio, and were at least 135% and 195% greater th an Dip n’ Grow at 1500 mgL-1 and all other treatments, respectively. Cuttings evaluated 4 WAP treated with KIBA at 6000 mgL-1 or Dip n’ Grow at 1500 mgL-1 had a similar growth response, which was better than all other treatments. Additiona lly, treating cuttings with KIBA at 3000 mgL-1 produced a 116% higher root : shoot ratio than at 1500 mgL-1, yet both were similar to the control. From H2 at 2 WAP cuttings treated with Dip n’ Grow at 1500 mgL-1 were similar to KIBA at 6000 mgL-1. Both treatments had at least a 151% higher root : shoot ra tio than cuttings treated with KIBA at 3000 mgL-1 or the control. Cuttings treated with KIBA at 1500 mgL-1 had a similar root : shoot ratio to KIBA concentrations of 3000 and 6000 mgL-1. Evaluations 4 WAP showed application of a rooting hormone at any concentration produced similar results that were at least 164% higher than the control. Cuttings planted from H3 had at least a 148% higher root : s hoot ratio 2 WAP when treated with KIBA at 3000 mgL-1 than all other treatments; all treatment s were similar to the control. Four weeks after planting, treatm ents of KIBA at 6000 mgL-1 or Dip n’ Grow at 1500 mgL-1 had higher root : shoot ratios than th e control and KIBA at 1500 or 3000 mgL-1. From H4, cuttings evaluated 2 WAP that we re treated with KIBA at 6000 mgL-1 or Dip n’ Grow at 1500 mgL-1 had a higher root : shoot ratio than cuttings treated with KIBA at 1500 mgL-1. KIBA at 3000 mgL-1 was similar to all treatments except the control. Root

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96 : shoot ratios were lower for the contro l than from any ot her rooting hormone concentration. When evaluated 4 WAP, cutti ngs treated with Dip n’ Grow at 1500 mgL1 had a higher root : shoot ratio than those treated with KIBA at 1500 mgL-1 or the control. KIBA at all concentr ations had similar root : shoo t ratios, however only KIBA at 3000 mgL-1 produced a higher ratio than th e control. KIBA at 6000 mgL-1 was similar to all treatments. Discussion During the first experiment rooting horm one applications to stem cuttings of allamanda generally did not influence per cent survival. Only KIBA at 3000 to 6000 mgL-1 improved rooting quality from the third ha rvest, otherwise all hormone treatments produced similar results to the control. Cuttings treated with higher rates of KIBA also tended to have more primary roots than the control, yet lateral root number was generally similar for all treatments. However, during evaluation of H3 rooting hormone application at any concentration improved lateral root num ber, when compared to the control. Results were similar for total cutting we ight and the root : shoot ratio amongst treatments, therefore suggesting no benefit of hormone applications ; however, the third harvest rooting hormone treatments produced greater root : shoot ratios when compared to the control. This occurrence, in add ition to improved visual rooting quality and increased numbers of primary and lateral roots, suggests application of KIBA at concentrations of 3000 to 6000 mgL-1 is desirable, especially after successive harvesting Dip n’ Grow at 1500 mgL-1 was also effective during the third harvest. Upon comparing observed parameters over time, KIBA at 3000 mgL-1 was the most consistent treatment, with similar results for visual rooting quality, number of prim ary roots, total cutting dry weight, and the root : shoot ratio throughout the study.

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97 During the second experiment, percent survival was generally high for all treatments and was not influenced by hormone application, similar to the first experiment. While the response to specific rooting hormones for either evaluation of each harvest was variable, certain trends em erged. Generally, all concentrations of rooting hormone applied to cu ttings tended to show an im provement in visual rooting quality when planted from the first and second harvests. Rooting quality tended to be lower for all treatments as the experiment progressed and differences amongst treatments were not significant by H3 and H4. This ma y be attributed to slower growth during natural short days, thus regardless of treatment, rooting quality was poor. The quantity of roots on untreated cuttings was similar at eac h harvest and tended to be lower than on hormone treated cuttings. When cuttings were planted from the third harvest, rooting was poor with respect to prim ary root number, yet results from all other harvests showed application of Dip n’ Grow at 1500 mgL-1 or KIBA at 3000 to 6000 mgL-1 generally showed improved rooting when compared to the control. The lower concentration of KIBA tended to be similar to the control. Lateral rooting was generally poor during the second study and was si milar with all treatments. Differences were not significant amongst tr eatments when considering total cutting dry weight, however, when consid ering the root : shoot ratio cu ttings treated with either Dip n’ Grow at 1500 mgL-1 or KIBA at 6000 mgL-1 showed greater ratios than the control. Better root development, as co mpared to shoot growth would be desired by growers for easier transplanting. When comparing rooting hormone treat ments during the first experiment, consistent rooting results occurred amongst harvests with KIBA at 3000 mgL-1. The

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98 second study showed better rooti ng with KIBA at 3000 to 6000 mgL-1 or Dip n’ Grow at 1500 mgL-1 as compared to the control or KIBA at 1500 mgL-1. During both studies, the third harvest generally exhibited the poor est rooting. Although it is unclear why this occurred, it may be attributed to aggressi ve harvesting of cut tings by not allowing optimal levels of carbohydr ates to accumulate. Conclusion Growers will want to consider the cost of rooting hormones and the labor costs associated with their application to the benefits achieved during propagation before deciding on a rooting hormone program. Currently, KIBA has limited commercial availability with a cost of $81.60 per 25 g (Sig ma-Aldrich Co., St. Lois, Mo.), while Dip n’ Grow is a commercially available produc t with a cost of $190.00 per 3.8 L (Morton’s Horticultural Products Inc., Mc Minnville, Tenn.). Based on consistent allamanda results with KIBA at 3000 mgL-1, its application at this concen tration would cost a propagator $9.79 per L (excluding labor), while Dip n’ Grow at 1500 mgL-1 would cost $5.02 per L. Bougainvillea glabra ‘California Gold’ Experiment 1 There was a harvest date x evaluation week interaction for bot h percent survival and rooting quality. Diffe rences in rooting quality were not significant amongst treatments, yet percent survival was greater fo r cuttings treated with any concentration of rooting hormone when compared to the c ontrol (data not shown) For both rooting quality and percent survival, differences be tween evaluation weeks were not significant for cuttings planted from H1, H3 and H4. Cuttings from H2 however, had a higher percent survival 7 WAP than at 5 WAP (98% vs 53%) (LSD=10, n=20). Rooting quality was also greater 7 WAP than at 5 WAP (data not shown).

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99 Table 4-2. Influence of rooting hormone a pplication on percent survival of allamanda cuttings evaluated 2 and 4 weeks after planting (WAP) from harvests (H) 1 to 4 during the warm season (Expt. 1) Percent survival (%) Treatment Concn (mgL-1) Week H1 H2 H3 H4 LSD Control 0 2 WAP 100 96 67 96 NS 4 WAP 100 100 83 83 NS NS NS NS NS Dip n' Grow 1500 2 WAP 92 96 96 100 NS 4 WAP 100 100 80 100 13 NS NS NS NS KIBA 1500 2 WAP 100 100 88 92 NS 4 WAP 100 100 100 79 ** 10 NS NS NS NS KIBA 3000 2 WAP 100 100 96 96 NS 4 WAP 100 96 96 96 NS NS NS NS NS KIBA 6000 2 WAP 100 100 100 96 NS 4 WAP 100 100 100 75 *** 8 NS NS NS 13 Significance 2 WAP NS NS NS LSD 22 Significance 4 WAP NS NS NS ** LSD 14 Source of variance Harvest date (H) *** Evaluation week (W) NS H x W Treatment (T) H x T *** W x T NS H x W x T NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt

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100 Table 4-8. Influence of rooting hormone a pplication on the visual rooting quality of allamanda cuttings evaluated 2 and 4 weeks after plan ting (WAP) from harvests (H) 1 to 4 of the cool season (Expt. 2). Rooting quality (1-5) Treatment Concn (mgL-1) Week H1 H2 H3 H4 LSD Control 0 2 WAP 1.9 1.7 1.0 2.0 0.7 4 WAP 3.3 3.5 2.5 3.7 0.9 NS NS ** LSD 0.9 0.7 Dip n' Grow 1500 2 WAP 3.8 3.2 1.1 3.7 *** 0.8 4 WAP 4.4 4.4 3.5 4.2 NS NS ** NS LSD 0.8 1.2 KIBA 1500 2 WAP 2.2 3.0 1.0 2.8 ** 1.0 4 WAP 3.8 4.4 2.7 3.4 ** 0.7 * NS LSD 1.0 0.8 1.2 KIBA 3000 2 WAP 3.0 3.1 1.1 3.1 *** 0.5 4 WAP 3.8 4.7 2.9 3.7 ** 0.9 * NS LSD 0.6 0.9 1.1 KIBA 6000 2 WAP 2.7 3.4 1.0 3.5 *** 0.4 4 WAP 3.8 4.7 3.5 3.7 NS * ** NS LSD 0.6 0.9 0.8 Significance 2 WAP ** NS LSD 0.8 0.7 0.9 Significance 4 WAP * NS NS LSD 0.6 0.6 Source of variance Harvest date (H) *** Evaluation week (W) *** H x W *** Treatment (T) *** H x T W x T NS H x W x T NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt

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101Table 4-10. Influence of rooting hormone on cutting root : shoot ratio of allamanda cuttings eval uated 2 and 4 weeks after pla nting (WAP) of harvests (H) 1 to 4 of the cool season (Expt. 2). Root : shoot ratio H1 H2 H3 H4 Treatment Concn (mgL-1) 2WAP4WAP 2WAP4WAP 2WAP4WAP 2WAP4WAP Control 0 0.10 0.29 0.09 0.28 0.10 0.23 0.12 0.22 Dip n' Grow 1500 0.22 0.39 0.32 0.52 0.14 0.40 0.28 0.41 KIBA 1500 0.10 0.28 0.23 0.46 0.12 0.24 0.20 0.28 KIBA 3000 0.15 0.33 0.18 0.46 0.21 0.28 0.23 0.39 KIBA 6000 0.30 0.42 0.27 0.47 0.12 0.40 0.29 0.32 Significance *** *** * * * LSDz 0.06 0.05 0.09 0.13 0.06 0.10 0.07 0.11 Source of variance Harvest date (H) *** Evaluation week (W) *** H x W *** Treatment (T) *** H x T ** W x T NS H x W x T ** NS, *, **, *** Significant at P 0.05, 0.01, 0,001. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt

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102 The harvest date x treatment interaction was not significant for number of primary roots or lateral roots, therefore the main effects will be presented. Differences in numbers of roots were not significant amongst treatments. Cuttings planted from H2, H3, and H4 had similar numbers of primary roots, 4.0 per cutting, which was at least 32% less than H1 (7.1) (LSD=1.8, n=20). Number of lateral roots was also greater from H1 (15.6) than H2 to H4, which rooted similarl y with 8.6 lateral root s per cutting (LSD=4.3, n=20). There was a harvest date x evaluation week and a harvest date x treatment interaction for cutting total dry weight. Cu ttings evaluated 7 WAP from H1 had a total dry weight of 355 mg per cutting, which wa s 113% greater than t hose evaluated 5 WAP (0.313 g) (LSD=25, n=20). Similarly, for H2, cuttings evaluated 5 WAP had a lower total dry weight than 7 WAP (data not shown) Differences in cutting total dry weight were not significant between evaluation w eeks for H3 (326 mg) or H4 (144 mg). Upon comparison of rooting hormone treatm ents, cuttings planted from H1 had a similar dry weight to the control when treated with Dip n’ Grow at 1500 mgL-1 or KIBA at 3000 mgL-1 (Table 4-11). Cuttings treated with KIBA at 3000 or 6000 mgL-1 weighed the same and were at least 12% lighter than the control. Cuttings treated with Dip n’ Grow at 1500 mgL-1 had similar dry weight to all treatments except lighter cuttings with KIBA at 6000 mgL-1. Differences in cutting total dry weight were not significant amongst treatments for H2 (227 mg) or H3 (325 mg). Cuttings planted from H4 had greater dry weight w ith any rooting hormone treatment when compared to the control (Table 4-11). Cuttings treated with KIBA at 3000 mgL-1 had greater dry weight

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103 than 6000 mgL-1, yet KIBA at 1500 mgL-1 was similar to all concentrations. Dip n’ Grow at 1500 mgL-1 also had similar total dry weight to all treatments except the control. When considering only untreated cuttings those planted from H1 and H3 had similar total dry weights and we re at least 133% greater than H2 and H4, which were also similar. Cuttings treated with Dip n’ Grow at 1500 mgL-1 had the same total dry weight when planted from H1, H3, and H4, while t hose planted from H2 were at least 27% lighter than all other harvests. Similarly, cuttings treated with KIBA concentrations from 1500 to 6000 mgL-1 had the lowest total dry weight at H2, while H1, H3 and H4 were similar and at least 143 % greater than H2. There was a harvest date x evaluation week interaction for the cutting root : shoot ratio. The main effects for treatment will be presented separately. Amongst harvest dates and evaluation weeks, cuttings treated with rooting hormone at all concentrations had a similar root : shoot ratio, and were at least 118% greater than the control (Data not shown). For cuttings planted from H1, t hose evaluated 5 WAP (0.12) had a 32% lower root : shoot ratio than at 7 WAP (0.36) (LSD=0.03, n=20). Similarly, cuttings evaluated from H2 had a greater root : shoot ratio at 7 WAP than at 5 WAP (Data not shown). For cuttings planted from H3, differences in r oot : shoot ratios were not significant between evaluation weeks (0.14). Cuttings planted from H4 had a root : shoot ratio of 0.15 seven WAP, which was 149% higher than at 5 WAP (0.10) (LSD=0.02, n=20). Experiment 2 There was a harvest date x evaluation week interaction for percent survival. Main effects for treatment were not significant. For rooting quality, the harvest date x evaluation week x treatment interactions were not significant, therefore the main effects are presented separately. Differences betw een evaluation dates were not significant for

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104 H1, H2, and H4; while cuttings planted from H3 had a greaer percent survival when evaluated 5 WAP than 7 WAP (68% vs 45%) (LSD=13 n=19). Differences amongst treatments for rooting quality were not signi ficant. Cuttings planted from H1, H2, H3, and H4 had visual rankings of 2.9, 3.3, 2.0, and 1.4, respectively (LSD=0.3, n=35). Table 4-11. Harvest date (H) by treatment in teraction for bougainvi llea ‘California Gold’ cuttings planted during the warm season (Expt. 1). Total cutting dry weight (mg) Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 362 247 328 220 *** 48 Dip n' Grow 1500 342 245 340 334 ** 56 KIBA 1500 317 220 312 330 *** 48 KIBA 3000 357 217 347 383 *** 58 KIBA 6000 293 205 300 296 ** 55 Significance NS NS *** LSD 40 67 NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt For both number of primary and lateral roots, the harvest date x treatment interaction was not significant, therefore the main effects are presen ted. Cuttings planted from H3 and H4 (1.2 and 0.5) had similar numb ers of primary roots, and were less than H1 and H2. Cuttings planted from H2 ha d 7.0 primary roots per cutting, which was 196% more than H1 (3.6) (LSD=1.4, n=17). Sim ilar to primary root number, lateral root number was highest for cuttings planted from H2 (14.9), while cutt ings from H3 and H4 (2.2 and 1.5) had at least 68% less latera l roots than H1 (7.0) (LSD=3.9, n=17). There was a harvest date x evaluation week x treatment interaction for cutting total dry weight and a harvest date x evaluation w eek interaction for the cutting root : shoot ratio. For cuttings planted from H1 and ev aluated 5 WAP, the c ontrol (363 mg) had a

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105 higher cutting total dry wei ght than those treated with Dip n’ Grow at 1500 mgL-1 (207 mg), while cuttings treated with KIBA at concentrations from 1500 to 6000 mgL-1 had similar dry weight to all other treatments (LSD=124, n=3). Differences in total cutting dry weight were not significant amongst treat ments 7 WAP. For cuttings planted from H2, those evaluated 5 WAP had similar total cutting dry weights when treated with a rooting hormone, all of which were at leas t 173% greater than the control (data not shown). Total cutting dry weight was simila r 7 WAP for cuttings treated with KIBA at 3000 or 6000 mgL-1 (350 or 366 mg), which was at least 155% higher than those treated with Dip n’ Grow at 1500 mgL-1, KIBA at 6000 mgL-1, or the control (226, 224, or 147 mg, respectively) (LSD=104, n=3). Differences in total cutting dry weight were not significant amongst treatments or between evaluation weeks from H3 or H4. There was a harvest date x evaluation week interaction for the cutting root : shoot ratio, however differences amongst treatments were not significant. For the first harvest, cuttings evaluated 5 WAP had a root : s hoot ratio of 0.32, while 7 WAP the ratio was 42% lower at 0.19 (LSD=0.07, n=16). Cuttings planted from the second harvest had a higher root : shoot ratio when evaluated 7 WAP than 5 WAP (0.27 vs 0.23) (LSD=0.03, n=15). Differences in the cutting root : shoot ratios were not significant between evaluation dates for either H3 or H4. Discussion During the first experiment, application of a rooting hormone improved survival, as compared to the control. For visual r ooting quality and number of both primary and lateral roots, rooting hormone treated cuttings performe d to the control; However, when considering the root : shoot ra tio, all rooting hormone treatme nts reflected greater ratios than the control. When comparing total dry weight, benefits of a rooting hormone

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106 application were demonstrated by increased dry weight when planted from H4, as compared to the control. Generally KIBA at 3000 mgL-1 or Dip n’ Grow at 1500 mgL-1 were shown to have the largest benef it in propagation of ‘California Gold’. Regardless of rooting hormone treatment duration of propagation time should be considered when rooting Bougainvillea ‘California Gold’. For H3, cuttings were shown to have nearly a 200% higher rate of surviv al at 7 WAP than 5 WAP; therefore it is recommended that propagators allow bougainvi llea cuttings to remain under mist for acceptable root development. Generally, cu ttings showed improved rooting quality 7 WAP when compared to evaluations at 5 WA P, opposite results may be due to disease pressure, nutrient leaching, and overall dec line in quality. Growers should consider application of fungicides to reduce the incide nce of disease and once weekly applications of fertilizer to cuttings. During the second experiment, percent surviv al, visual rooting quality, primary and lateral root number, and the cutting root : s hoot ratios were all si milar amongst rooting hormone treatments and were not shown to e nhance rooting quality and performance, as compared to the control. Differences in to tal cutting dry weight were less apparent as time progressed. Over time, reduced rooting occurred as the duration of the study progressed into fall, except for cuttings planted from H2 wh ich tended to root better than H1, H3, and H4;. This reduction in rooti ng performance could be the re sult of physiological changes within the cutting as a result of flowering unde r naturally shorter days in fall or possibly diminishing nutritional status of cuttings and stock plants.

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107 Conclusion Cuttings of ‘California Gold’ rooted with increased quality during the first experiment (warm season) when rooting hormone was applied, however treatment differences were not significant during th e fall (cool) growing season. Regardless of rooting hormone treatment, overall rooting performance decreased in fall. Growers will want to consider planting mo re cuttings during the summer months and application of rooting hormone to hasten th e rooting process and improve quality during those months. Currently, KIBA has limited commercial av ailability with a cost of $81.60 per 25 g (Sigma-Aldrich Co., St. Lois, Mo.), while Di p n’ Grow is a commercially available product with a cost of $190.00 per 3.8 L (M orton’s Horticultural Products Inc., McMinnville, Tenn.). Application of Di p n’ Grow or KIBA mixed at 1500 mgL-1 would cost a propagator $5.02 and $4.90 per L (excluding labor), respectively. Bougainvillea glabra ‘Helen Johnson’ Experiment 1 There was a harvest date x evaluation w eek x treatment interaction for percent survival. Differences in percent survival were not significant amongst treatments 5 or 7 WAP for H1 and H2. For cuttings planted from H3 and evaluated 5 WAP, all rooting hormone treatments had a 98% survival, wh ich was greater than the control (83%) (LSD=7, n=4). At 7 WAP, differences in pe rcent survival were not significant amongst treatments (90%). From H4, cuttings evalua ted 5 WAP had greater percent survival with application rooting hormone than when left untreated (data not shown). Differences in percent survival were not significant amongst treatments when evaluated 7 WAP. For untreated cuttings, those evaluated at both 5 and 7 WAP had similar percent survival, except those planted from H2, wh ich had greater survival 5 WAP than at 7

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108 WAP (100% vs 75%) (LSD=15, n=4). For cutt ings treated with Dip n’ Grow at 1500 mgL-1 or KIBA at 3000 mgL-1, differences between cutti ngs evaluated 5 and 7 WAP were not significant for all harvest dates. Cuttings treated with KIBA at 1500 or 6000 mgL-1 had greater percent surviv al from H1 when evaluate d 5 WAP as compared to 7 WAP (data not shown); however, differen ces between evaluation weeks were not significant for H2, H3, or H4. There was a harvest date x treatment intera ction for rooting qual ity. Main effects for evaluation week will be presented separa tely. Amongst all harvest dates, cuttings evaluated 7 WAP had an averag e rooting quality of 2.4, which was greater than 5 WAP at 2.0 (LSD=0.2, n=80). For H1, H2, and H4 di fferences in rooting quality were not significant amongst treatments, with an average rooting quality of 1.6, 2.4, and 1.7, respectively. For cuttings planted from H3, r ooting quality of cuttings treated with KIBA at 3000 or 6000 mgL-1 (2.6 or 3.0, respectively) had sim ilar rooting quality as compared to the control (2.5), while Dip n’ Grow at 1500 mgL-1 (3.8) was similar to KIBA at 1500 mgL-1 (3.3) (LSD=0.7, n=8). Additionally, cu ttings treated with KIBA at 1500 mgL-1 had greater rooting qua lity than at 3000 mgL-1, yet both were similar to KIBA at 6000 mgL-1. When considering only untreated cuttings, those planted from H3 (2.5) had greater rooting quality than all other harvests, excep t H2 (1.9). Untreated cuttings planted from H2 were similar in rooting quality to both H3 and H4 (1.6), while H1 (1.2) was lower than all harvests except H4 (LSD= 0.7, n=8). Cuttings treated with Dip n’ Grow at 1500 mgL-1, from H1 (1.5) had lower rooting quality than all harvest dates, except H4 (2.0), which was similar to H2 (2.7). Cuttings pl anted from H3 (3.8) ha d the highest rooting

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109 quality (LSD=0.9, n=8). When considering only cuttings treated with KIBA at 1500 mgL-1, those planted from H3 (3.3) had greate r rooting quality than all other harvest dates. While H2 (2.2) was greater than H4 (1.3), cuttings planted from H1 (1.8) had similar rooting quality to both H2 and H4 (LSD=0.7, n=8). For cuttings treated with KIBA at 3000 mgL-1, H2 and H3 (2.6) had similar root ing quality, yet were greater than H4 (1.6). Cuttings planted from H1 (1.9) we re similar to all othe r treatments. (LSD=0.8, n=8). Rooting quality of cutti ngs treated with KIBA at 6000 mgL-1 was similar when planted from H2 and H3, both were greater than H4 and H1, which were also similar (data not shown). The harvest date x treatment interaction was not significant for either primary or lateral root number, therefore the main effect s will be presented. When considering the number of primary roots, cuttings treate d with Dip n’ Grow or KIBA at 1500 mgL-1 (2.6 or 1.9, respectively) had a similar number of primary roots to the control (1.6) (LSD=1.5, n=16). Additionally, KIBA at 1500 mgL-1 was similar in rooting to KIBA at 3000 mgL1 (7.9); concentrations of 3000 to 6000 mgL-1 produced at least 208% more primary roots than the control. Cuttings trea ted with Dip n’ Grow at 1500 mgL-1 (2.6) had a similar number of primary roots to all treatments. Amongst treatments, H2 (4.0) and H3 (4.2) produced a similar number of primary roots pe r cutting, which were at least 253% greater in number than from H1 (0.6) or H4 (1.6) (LSD=1.4, n=20). For numbe r of lateral roots, cuttings treated with KIBA at 3000 or 6000 mgL-1 (7.9 or 9.4) were similar, and had at least 231% more lateral root s than either the control (2.2) or KIBA at 1500 mgL-1 (3.4), while cuttings treated with Dip n’ Grow at 1500 mgL-1 (5.8) were similar to all treatments (LSD=4.4, n=16). Amongst treatment s, cuttings from H3 produced 9.2 lateral

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110 roots per cutting, which was at least 213% more than H1 or H4 (3.3 or 4.3), while H2 (6.2) was similar to all harvests (LSD=4.0 n=20). The harvest date x treatment interacti on was not significant for total cutting dry weight; therefore the main effects will be presented separately. Differences in total cutting dry weight were not significant amongst treatments. Cuttings planted from H1 (213 mg) and H2 (199 mg) had similar total cu tting dry weight, and were at least 15% less and 120 % greater than H3 and H4, re spectively (250 or 166 mg) (LSD=23, n=40). Table 4-12. Harvest date (H) by treatment intera ction for root : shoot ratio (root weight + shoot weight) of ‘Helen Johnson’ bouga invillea cuttings planted during the warm season (Expt. 1). Root : shoot ratio Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 0.12 0.18 0.20 0.17 NS Dip n' Grow 1500 0.13 0.32 0.53 0.23 ** 0.18 KIBA 1500 0.09 0.22 0.29 0.15 ** 0.10 KIBA 3000 0.13 0.30 0.21 0.17 ** 0.08 KIBA 6000 0.20 0.34 0.28 0.18 0.11 Significance * NS LSD 0.07 0.12 0.12 NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt There was a harvest date x evaluation week, harvest date x treatment, and evaluation week x treatment interaction for cu tting root : shoot ratio. For H1, cuttings treated with KIBA at 6000 mgL-1, had a greater root : s hoot ratio than all other treatments (Table 4-12). Cutti ngs treated with KIBA at 6000 mgL-1 or Dip n’ Grow at 1500 mgL-1 had a higher ratio than the control from H2. Untreated cuttings had a similar root : shoot ratio when tr eated with KIBA at 1500 mgL-1. Cuttings treated with KIBA at 3000 mgL-1 had a similar root : shoot ratio to all other treatments, whil e Dip n’ Grow at

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111 1500 mgL-1 had a similar root : shoo t ratio to all concentrations of KIBA. Harvest 3 cuttings had a higher root : shoot ratio when treated with Dip n’ Grow at 1500 mgL-1, when compared to all other treatments. Cutti ngs treated with all co ncentrations of KIBA were similar to the control. Differences in cutting root : ratio were not significant amongst treatments for cuttings planted fr om H4 (0.18). When considering only untreated cuttings, all harves t dates produced similar root : shoot ratios of 0.13. For cuttings treated with Dip n’ Grow at 1500 mgL-1, those planted from H3 had greater root : shoot ratio than all other harv est dates. While H2 had greate r root : shoot ratio than H1, H4 ratios were similar to both. KIBA at 1500 mgL-1 had a greater root : shoot ratio from H3 than H4. Cuttings planted from H1 were similar in root : shoot ratio only to H4. When considering only cuttings treated with KIBA at 3000 mgL-1, H2 had larger ratio than all other harvests. Harvest 3 cuttings were greater than H1, and both H1 and H3 were similar to H4. Cuttings treated with KIBA at 6000 mgL-1 had greater root : shoot ratio from H2 than H1 and H4. Cuttings from all harvests were similar in root : shoot ratio to H3. Differences between evaluation weeks were not significant amongst treatments for H1; however, from H2, H3, and H4 cuttings evaluated 7 WAP had larger root : shoot ratios than at 5 WAP (data not shown). Am ongst all harvest dates, cuttings evaluated 5 WAP showed at least a 145% higher root : shoot ratio with KIBA at 6000 mgL-1 (0.24) or Dip n’ Grow at 1500 mgL-1 (0.21) than KIBA at 1500 mgL-1 (0.15) or the control (0.13) (LSD=0.06, n=16). Cuttings treated with KIBA at 1500 or 3000 mgL-1 were similar to the control, while those treated with Dip n’ Grow at 1500 mgL-1 were similar to both KIBA at 3000 or 6000 mgL-1. At 7 WAP cuttings treated with Dip n’ Grow at

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112 1500 mgL-1 had a higher root : shoot ratio than all other treatments, while KIBA at all concentrations were similar to the control (data not shown). Experiment 2 There was a harvest date x treatment interaction for both percent survival and rooting quality. Additionally there was a harvest date x ev aluation week interaction for percent survival. Main effects for evaluation week will be presented separately for rooting quality. For cuttings planted from H1, differences in percent survival were not significant amongst treatments (70%). Cuttings planted from H2 had a similar percent survival when treated with KIBA at 3000 or 6000 mgL-1 or Dip n’ Grow at 1500 mgL-1 (93%, 98%, and 92%, respectively) all of which were greater than the control or KIBA at 6000 mgL-1 (LSD=18, n=8). From H3, the control (50%) was similar to all treatments, as was KIBA at 1500 mgL-1 (60%). Cuttings treated wi th Dip n’ Grow at 1500 mgL-1 (48%) had a similar percent survival to all treatments except KIBA at 6000 mgL-1 (71%), which was at least 150% greater than cu ttings treated with KIBA at 3000 mgL-1 (43%) (LSD=22, n=6). Differences in percent surviv al for cuttings planted from H4 were not significant amongst treatments. Differences am ongst harvest dates were not significant for untreated cuttings (68%) and KIBA at 6000 mgL-1 (72%). For cuttings treated with either Dip n’ Grow or KIBA at concentrations of 1500 mgL-1, those planted from H2 and H4 had similar percent survival values. Both H2 and H4 survived better than H1 and H3 (data not shown). KIBA at 3000 mgL-1 produced similar survival rates from H2 and H4 (98% and 87%), yet was at least 19% and 51% greater than H1 (71%) and H3 (43%), respectively (LSD=17, n=6). For cuttings planted from H1, H2, a nd H4, those evaluated 7 WAP survived similarly as compared to 5 WAP (data not shown). Cuttings planted from H3 and

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113 evaluated 7 WAP had a 63% survival amongs t treatments, which was greater than 5 WAP (41%) (LSD=14, n=16). Comparatively, cuttings evaluated at 5 WAP or 7 WAP from H1, H2, or H4 had similar survival rate s, while H3 had a lower survival percentage at both 5 and 7 WAP. Rooting quality of cuttings planted from H1 was similar amongst treatments with a mean value of 2.7. From H2, the rooting qua lity of cuttings treated with KIBA at 6000 mgL-1 was similar to the control, while both were less than all other treatments (Table 413). Dip n’ Grow at 1500 mgL-1 and KIBA at 1500 or 3000 mgL-1 all had similar rooting quality values. Cuttings planted fr om H3 had similar rooting quality to the control when treated with KIBA at 1500 or 6000 mgL-1. Cuttings treated with Dip n’ Grow at 1500 mgL-1 had greater rooting quality than KIBA at 6000 mgL-1, however both were similar to KIBA at 3000 mgL-1. KIBA at 1500 mgL-1 was similar to 6000 mgL-1, yet was lower than all other hormone treat ments. For H4, cuttings treated with Dip n’ Grow at 1500 mgL-1 had greater rooting quality than all other treatments except KIBA at 1500 mgL-1, which was similar to all treatments. Cuttings had similar rooting quality values when untreated or treated with KIBA at 3000 or 6000 mgL-1. KIBA at all concentrations produced similar rooting quality values. When considering only cuttings treated with KIBA at 1500 mgL-1, 6000 mgL-1, or the control, those planted from H1, H2, and H4 produced similar r ooting quality values, all of which were greater than H3 (Table 4-13). Cuttings treated with Dip n’ Grow at 1500 mgL-1 had similar rooting quality for H2 and H4, and both were grea ter than H1 and H3. KIBA at 3000 mgL-1 had similar results from H1 and H4, yet H2 and H3 had higher and lower quality ratings than

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114 H1 and H4, respectively. Am ongst all treatments, cuttings evaluated 7 WAP had greater rooting quality than t hose evaluated 5 WAP (2.8 vs 2.5) (LSD=0.2 n=65). There was a harvest date x treatment interaction for both primary and lateral root number. Differences in number of primar y roots were not significant amongst treatments for H1, H2, and H3, with 4.1, 6.1, and 0.7 primary roots produced per cutting, respectively (Table 4-14). For cuttings planted from H4, a ll treatments were similar to the control, except those treate d with Dip n’ Grow at 1500 mgL-1, which had at least 193% more roots than all othe r treatments. When consid ering only untreated cuttings, H2 had more primary roots than H3, while H1 and H4, were similar to all harvest dates. When cuttings were treated with Dip n’ Grow at 1500 mgL-1, H2 had the same number of primary roots as both H4 and H1, while H1 was similar to H3. Harvest 4 had a greater number of primary roots than both H1 and H 3. Differences in primary roots were not significant amongst treatments for cutti ngs treated with KIBA at 1500 mgL-1. When treated with KIBA at 3000 mgL-1, cuttings planted from H2 had more primary roots than all other harvest dates. Cuttings treated with KIBA at 6000 mgL-1 had more primary roots when planted from H2 than from H4, however both were similar to H1. Additionally, cuttings planted from H3 had a lower number of primary roots than H1, yet both H1 and H3 were similar to H4. Differences in lateral root number were not significant amongst treatments for H1 (9.2) and H3 (0.7). For cuttings planted from H2, all treatments, except those treated with KIBA at 3000 mgL-1, had a similar number of late ral roots, including the control (Table 4-15). For cuttings planted from H 4, all treatments, except Dip n’ Grow at 1500 mgL-1, had a similar number of lateral roots when compared to the control. When

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115 considering only untreated cuttings, KIBA at 1500 mgL-1, or KIBA at 6000 mgL-1, differences in lateral root number were not significant amongst harvest dates. For cuttings treated with Dip n’ Grow at 1500 mgL-1 or KIBA at 3000 mgL-1 H2 had the same number of lateral roots as both H4 and H1, while H1 was similar to H3. Harvest 4 had a greater number of lateral roots than both H1 and H 3. This was a similar trend to primary root number. Cutting total dry weight was 300 mg per cutting, which was not significantly different amongst treatments or harvest dates. There was a harvest date x evaluation week and a harvest date x treatment intera ction for the cutting root : shoot ratio. Differences in cutting root : shoot ratio we re not significant amongst treatments for those planted from H1 and H3. Cuttings planted fr om H2 had a root : shoot ratio of 0.57 when treated with Dip n’ Grow at 1500 mgL-1, which was similar to all other hormone treatments, except KIBA at 6000 mgL-1 (0.38) or the control (0.28) (LSD=0.15, n=8). Additionally, KIBA at all c oncentrations had similar ro ot : shoot ratios, although concentrations of 1500 to 3000 mgL-1 produced larger ratios than the control (0.46 vs 0.28). For cuttings planted from H4, those treated with a rooting hormone at any concentration had a similar root : shoot ra tio, which was at least 145% higher than the untreated cuttings (data not shown). For each treatment, cuttings planted from H2 had a greater root : shoot ratio than from all other harvest dates. When considering evaluation week, differences in cutting r oot : shoot ratio were not si gnificant betwee n 5 and 7 WAP for harvests H1, H3, and H4, while cuttings evaluated 7 WAP (0.48) from H2, had a 128% higher root : shoot ratio than at 5 WAP (0.38) (LSD=0.10, n=20).

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116 Table 4-13. Harvest date (H) by treatment in teraction for visual rooting quality (1-5; 1=poor, 5=best) of bougainvillea ‘Helen J ohnson’ cuttings planted during the cool season (Expt. 2). Rooting quality Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 2.4 3.0 1.2 2.5 *** 0.6 Dip n' Grow 1500 2.2 3.4 2.0 3.5 *** 0.2 KIBA 1500 3.0 3.2 1.2 3.1 *** 0.8 KIBA 3000 2.8 4.1 1.8 2.7 *** 0.7 KIBA 6000 2.9 3.3 1.4 2.7 *** 1.0 Significance NS ** ** LSD 0.7 0.5 0.6 NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt Table 4-14. Harvest date (H) by treatment in teraction for number of primary roots of bougainvillea ‘Helen Johnson’ cuttings planted during the cool season (Expt. 2). Number of primary roots Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 2.9 5.4 0.0 2.1 3.7 Dip n' Grow 1500 4.0 8.7 1.1 9.5 5.5 KIBA 1500 4.2 8.2 0.8 4.9 NS KIBA 3000 4.0 12.5 1.5 3.5 *** 3.2 KIBA 6000 5.4 6.7 0.3 2.1 4.5 Significance NS NS NS LSD 4.2 NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt

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117 Table 4-15. Harvest date (H) by treatment in teraction for number of lateral roots of bougainvillea ‘Helen Johnson’ cuttings planted during the cool season (Expt. 2). Number of lateral roots Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 6.2 8.6 0.0 9.8 NS Dip n' Grow 1500 6.1 14.1 1.3 25.0 ** 11.8 KIBA 1500 12.1 11.6 0.0 9.3 NS KIBA 3000 9.7 29.0 2.0 11.6 *** 9.8 KIBA 6000 12.0 14.4 0.0 4.0 NS Significance NS NS ** LSD 10.6 9.5 NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt

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118 Discussion During the first experiment, the application of a rooting hormone at H1 and H2 did not improve percent survival. By the third ha rvest however, all concen trations of rooting hormones improved survival, as compared to the control when evaluated 5 WAP, but not at 7 WAP; results were similar for the four th harvest. This suggests the benefit to application of rooting hormone is achieved through hastening early establishment of cuttings, but is less important for surv ival as duration under mist increases. In most instances application of a rooti ng hormone did not impr ove visual rooting quality, when compared to the control When planted from the third harvest however, cuttings showed improved rooting with app lications of Dip n’ Grow or KIBA at 1500 mgL-1 when compared to the control. Primar y and lateral root numbers were improved when cuttings were provided KIBA at concentrations of 3000 to 6000 mgL-1. This enhancement of roots with hormones shoul d provide benefit to propagators when transplanting bougainvillea cuttings with fragil e root systems. This research also demonstrated the importance of measuring root number vs. visual estimation of quality. Total cutting dry weight was similar amongst treatments, while the root : shoot rati, was improved early in the experiment with applications of Dip n’ Grow or KIBA at concentrations of 1500 and 6000 mgL-1, respectively. When cuttings were evaluated from H4, all treatments had similar root : s hoot ratios. When ratios were pooled amongst harvest dates to investigate evaluation date treating cuttings with Dip n’ Grow at 1500 mgL-1 was shown to benefit the root : shoot ratio 7 WAP. During the second experiment, percent su rvival was generally improved when cuttings were provided Dip n’ Grow at 1500 mgL-1 or KIBA at 3000 to 6000 mgL-1.

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119 Percent survival was similar at both evaluation fates except at H3 when cuttings evaluated 7 WAP survived better than 5 WAP. Rooting quality was similar for all treatments at the first harvest, but for H2 to H4, rankings were generally improved by app lication of Dip n’ Grow at 1500 mgL-1 or KIBA at 1500 to 3000 mgL-1. From H4 primary root number was improved with Dip n’ Grow at 1500 mgL-1 as compared to the control, otherwise root number was similar amongst treatments from all harvests. Lateral root number was similar amongst treatments when planted at all harvests, ex cept H3 in which rooting was improved with KIBA at 3000 mgL-1, as compared to the control. Gene rally applications of Dip n’ Grow at 1500 mgL-1 or KIBA at 1500 to 3000 mgL-1 were shown to have greater root : shoot ratios than the control, however total cutting dry weights was similar amongst treatments. In both experiments differences between cu ttings evaluated 5 and 7 WAP tended to be similar with regards to percent survival It is therefore re commended to remove cuttings from mist early (5 WAP) during th e active growth season, rather than allowing extensive time in propagation. All other pa rameters showing improvement 7 WAP, as compared to 5 WAP would othe rwise indicate growth and development with time, and may be enhanced with once-weekly fertilizer applications. Conclusion Application of rooting hormones to stem cuttings of ‘Helen Johnson’ generally improved rooting performance as compared to the control, with Dip n’ Grow at 1500 mgL-1 or KIBA at 3000 mgL-1, producing the best results. Currently, KIBA has limited commercial availability with a cost of $81.60 per 25 g (Sigma-Aldrich Co., St. Lois, Mo.), while Dip n’ Grow is a commercially available product w ith a cost of $190.00 per 3.8 L (Morton’s Horticultural Products Inc., Mc Minnville, Tenn.). Application of Dip n’

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120 Grow or KIBA mixed at 1500 mgL-1 would cost a propagator $5.02 and $4.90 per L (excluding labor), respectively. Propagators should consider cost of application and benefits achieved in rooting performance be fore deciding on a roo ting hormone program. Mandevilla splendens ‘White’ Experiment 1 The harvest date x evaluation week x treatment interaction was not significant for percent survival; therefore the main effects will be presented. Differences in percent survival were not significant amongst treatment s. Mandevilla cuttings planted from H3 (81%) had greater percent survival than eith er H1 or H2 (59%) (LSD=10, n=40). When comparing evaluation dates, those cuttings ev aluated 6 WAP (77%) had a higher survival percentage than at 4 WAP (55%) (LSD=8, n=60). There was a harvest date x evaluation w eek interaction for rooting quality. Main effects for treatment will be presented separately. Cuttings planted from H1 had an average visual ranking of 1.7 and 3.8 for evaluations 4 and 6 WAP, respectively (LSD=0.6, n=20). Results from H2 and H3 also had greater rooting quality 6 WAP as compared to 4 WAP (data not shown). Differen ces in rooting quality were not significant amongst treatments. The harvest date x treatment interaction was not significant for number of primary or lateral roots, therefore the main effect s are presented separately. Differences in primary or lateral root number were not significant amongst tr eatments. Rooted cuttings from the three harvests of mandevilla had 1.9, 3.4 and 5.3 primary roots per cutting respectively; each harvest had a greater num ber of roots than the previous (LSD=1.4 n=20). Differences in lateral root number were not significant amongst harvest dates.

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121 Differences in either total cutting dry we ight or the root : shoot ratio were not significant amongst treatments. Both total dry weight and root : shoot ratio were greater when evaluated 6 WAP than cuttings eval uated 4 WAP (data not shown). Cuttings planted from H1 had a total dry weight of 424 mg, which was 164% and 146% higher than H2 (258 mg) and H3 (290 mg), respectiv ely (LSD=28, n=40). Di fferences in cutting total dry weight were not significant between harvests. Experiment 2 The harvest date x treatment interaction was not significant for percent survival; therefore the main effects will be presented. All mandevilla cuttings planted during the cool season had similar percent survival as co mpared to the control, while those treated with KIBA at 1500 or 6000 mgL-1 (82% and 79%, respective ly) had a greater survival rate than those treated with Dip n’ Grow at 1500 mgL-1 (67%) (LSD=9, n=32). Additionally, all concentr ations of KIBA had similar percen t survival. Cuttings from H1 and H2 expressed an 85% survival rate, whic h was greater than H3 (75%) and H4 (60%) (LSD=8, n=40). There was a harvest date x treatment inte raction for rooting quality. The main effects for evaluation week will be presented separately. For H1, cuttings treated with Dip n’ Grow at 1500 mgL-1 had greater rooting quality th an all other rooting hormone treatments, which were all similar (Table 416). For cuttings planted from H2, all treatments were similar to the control except KIBA at 1500 mgL-1. All rates of KIBA had similar rooting quality, while those treated with KIBA at 1500 or 3000 mgL-1 had greater root quality ratings than cuttings treated with Dip n’ Grow at 1500 mgL-1. Rooting quality was similar for all treatments wh en cuttings were planted from H3 or H4. When considering untreated cuttings or KIBA at 6000 mgL-1, results from each harvest

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122 planting were similar, while cuttings treated with Dip n’ Grow at 1500 mgL-1 had higher rooting quality from H1 as compared to all ot her harvest dates. For cuttings treated with KIBA at 1500 mgL-1, H1 had similar rooting quality to all harvest dates, although H2 was greater than H3 and H4. Cuttings treated with KIBA at 3000 mgL-1 had similar rooting quality from all harvests except H4, which was the lowest. Amongst all treatments, cuttings evaluated 6 WAP had a r oot quality ranking of 3.2, which was higher than cuttings evaluated 4 WAP (2.7) (LSD=0.2, n=79). Table 416. Harvest date by treatment interac tion for visual rooting quality (1-5; 1=poor, 5=best) of mandevilla cuttings planted during the cool season (Expt. 2). Rooting quality Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 2.9 2.8 2.5 2.7 NS Dip n' Grow 1500 4.0 2.7 3.1 2.5 ** 0.8 KIBA 1500 3.0 3.7 2.5 2.6 0.8 KIBA 3000 3.0 3.4 2.8 1.9 ** 0.7 KIBA 6000 2.9 3.3 3.1 3.0 NS Significance * NS NS LSD 0.7 0.6 NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt The harvest date x treatment interaction was not significant for number of primary or lateral roots; therefore the main effects will be presented. Differences in primary root number were not significant amongst harvests or treatments. Number of lateral roots was similar for all harvests (6.7) except H3 (0.3), which was at least 94% lower than other harvest dates (LSD=4.8, n=20). There was a harvest date x evaluation week interaction for both cutting total dry weight and the root : shoot ratio. Additio nally, there was a harvest date x treatment

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123 interaction for cutting total dry weight. Differences in total cutting dry weight were not significant amongst treatments for all harves t plantings during the cool season. When considering Dip n’ Grow at 1500 mgL-1 or KIBA at 6000 mgL-1 the total cutting dry weight was similar for cuttings planted from each harvest. With untr eated cuttings, total cutting dry weight tended to increase as th e season progressed (data not shown). Cuttings treated with KIBA at 1500 mgL-1 had lower dry weight when planted from H1 (204 mg) than from H3 (293 mg), yet H2 (240 mg) and H4 (245 mg) were similar to all harvests (LSD=59, n=8). When considering only cu ttings treated with KIBA at 3000 mgL-1, those planted from H1 (160 mg) had similar to tal dry weight to H4 (202 mg), while both H1 and H4 were at least 24% less than H2 (265 mg) and H3 (281 mg) (LSD=55, n=8). Cuttings planted from H1 and evaluated 4 WAP (212 mg) had 140% greater total dry weight than those evaluated 6 WAP (151 mg ) (LSD=33, n=20). For cuttings planted from H2, H3, and H4, cutting total dry weight was similar at both 4 a nd 6 WAP. For root : shoot ratio, cuttings evaluated 6 WAP we re 256%, 168%, 148%, and 189% greater than cuttings evaluated 4 WAP from harvests H1, H2, H3, and H4, respectively (data not shown). Discussion During the first experiment, rooting performance of mandevilla cuttings was generally similar between the untreated cuttings and those provided with rooting hormones, therefore applicati on of rooting hormone was not shown to be beneficial during the warm season. When comparing ev aluation dates, cuttings evaluated 6 WAP tended to have greater survival than at 4 WAP, which suggests the importance of allowing plants to remain under mist for an ex tended period. Decline in overall quality of propagules existed as length of time under mist increased, therefore application of

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124 fungicide or a once-weekly fertilizers during propagati on may be beneficial to propagators wishing to produce mandevilla liners. During the second experiment percent surv ival was similar for the control and hormone-treated cuttings. Rooting hormones tended to improve rooting quality as compared to the control early in the fall gr owing season. Cuttings treated with Dip n’ Grow at 1500 mgL-1 or KIBA at 1500 mgL-1, were shown to improve rooting at H1 and H2, respectively. As the study progressed, appl ication of rooting hormone to cuttings produced similar rooting quality values to th e control. Both primary and lateral root numbers, along with total dry weight and th e root : shoot ratio, were not significant amongst treatments. When comparing each treatment over time, KIBA at 6000 mgL-1 was consistent from each harvest with respec t to rooting quality and total dry weight, therefore this treatment may have merit wh en evaluating other propagation factors with mandevilla. Conclusion Over both studies, propagation of Mandevilla splendens ‘White’ was generally similar amongst all treatments. However, application of a roo ting hormone during the second study (cool season) produced better root ing quality values with Dip n’ Grow or KIBA at 1500 mgL-1, when compared to the control. KIBA at 6000 mgL-1 was consistent in rooting quality throughout the study, when co mpared to other treatments and may be an important characteristic for investigation of stock plant parameters in future research, however bot h Dip n’ Grow at 1500 mgL-1 and KIBA at 1500 mgL-1 would be economically feasible. There was also a trend toward increased propagation success during the cooler growing season. Fu rther investigation should be conducted to more fully define the seasonal effects when propagating mandevilla. Growers will want

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125 to compare cost of application of rooting horm one to benefits achieve d in rooting quality. Currently, KIBA has limited commercial av ailability with a cost of $81.60 per 25 g (Sigma-Aldrich Co., St. Lois, Mo.), while Di p n’ Grow is a commercially available product with a cost of $190.00 per 3.8 L (M orton’s Horticultural Products Inc., McMinnville, Tenn.). Application of Di p n’ Grow or KIBA mixed at 1500 mgL-1 would cost a propagator $5.02 and $4.90 per L (exclu ding labor), respectively. Propagators should consider cost of application and benef its achieved in rooting performance before deciding on a rooting hormone program. Nerium oleander ‘Dwarf Salmon’ Experiment 1 There was a harvest date x treatment interaction for both percent survival and rooting quality. Main effects for evaluation week will be presented separately. Differences in percent survival were not significant amongst treatments for cuttings planted from H1, H2, and H4. For cuttings pl anted from H3, percent survival was similar for all hormone treated cuttings (95%), wh ich were greater than the control (73%) (LSD=11, n=8). For all treatments, cuttings planted from H4 had the lowest survival percentage, while H1 through H3 were simila r, except those treated with KIBA at 6000 mgL-1, which resulted in low survival ra tes from H1 (data not shown). Cutting rooting quality from H1 improved when treated with a rooting hormone except for KIBA at 3000 mgL-1, which was similar to the control (Table 4-17). For cuttings planted from H2, visual rooting qua lity was similar for all treatments. When cuttings were planted from H3, quality ratings of cuttings treated with KIBA at 3000 or 6000 mgL-1 were the only treatments to show improved rooting, as compared to the control. While all KIBA concentrations had similar rooting quality, KIBA at 6000 mgL-

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126 1 had better rooting quality th an Dip n’ Grow at 1500 mgL-1, which was similar to all other treatments. For cuttings planted from H4 rooting qua lity was generally poor. All treatments were similar to the control, except Dip n’ Grow at 1500 mgL-1, which was lower. While KIBA at 3000 mgL-1 was similar to the control, it expressed better rooting quality than KIBA or Dip n’ Grow at 1500 mgL-1. When considering the rooting response to each treatment over time, both the untreated cuttings and KIBA at 3000 mgL-1 had similar results when planted from H1 to H4. Cuttings treated with Dip n’ Grow at 1500 mgL-1 or KIBA at 1500 or 6000 mgL-1, had reduced rooting quality from H4, when compared to any other harvest date. Differences in percent survival were not significant between evaluation dates; howev er, cuttings evaluated 6 WAP had better rooting quality than t hose evaluated 4 WAP (3.2 vs 2.9) (LSD=0.3, n=78). Table 4-17. Harvest date (H) by treatment in teraction for visual rooting quality (1-5; 1=poor, 5=best) of oleander cuttings planted during the warm season (Expt. 1). Rooting quality Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 2.8 3.1 3.1 2.5 NS Dip n' Grow 1500 3.9 3.5 3.3 0.4 *** 0.7 KIBA 1500 3.9 3.4 3.3 0.4 *** 0.9 KIBA 3000 3.5 3.7 3.7 2.9 NS KIBA 6000 3.8 3.9 3.8 1.9 1.5 Significance NS * LSD 0.8 0.5 1.5 NS, *, *** Nonsignificant or significant at P 0.05 or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt The harvest date x treatment interaction was not significant for number of primary roots, yet results for lateral root number were significant. The main effects for primary roots will be presented separa tely. Differences in the numbe r of primary roots were not

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127 significant amongst treatments. When comparing harvest dates, cuttings planted from H4 had the lowest number of primary roots and we re at least 74% lower than other harvest dates (data not shown). For lateral root numbe r, cuttings planted from H1 had more roots than the control when treated with ei ther Dip n’ Grow or KIBA at 1500 mgL-1 (Table 418). Cuttings treated with KIBA at 3000 to 6000 mgL-1 produced a similar number of lateral roots, when compared to all treatme nts. When planted from H2, lateral root number was greater when cuttings were treated with KIBA at 6000mgL-1 than any other treatment, except KIBA at 3000 mgL-1. Cuttings treated with KIBA at 3000 mgL-1 had a similar number of lateral roots to all treatments, including the control. Differences in lateral root number were not significant amongst treatments for cuttings planted from H3 or H4. When treated with Dip n’ Grow at 1500 mgL-1 or left untreated, cuttings planted from all harvests had a similar number of late ral roots. Cuttings treated with KIBA at 1500 mgL-1 had the greatest quantity of lateral roots when planted from H1. As the duration of the experiment progressed, lateral root numbers decreased, as H2 was greater than H4, yet both H2 and H4 were similar to H3. For cuttings trea ted with KIBA at 3000 mgL-1, H2 had more lateral roots when compared to H3 or H4, yet H1 was similar to those planted from all harvests. Similarl y, cuttings treated with KIBA at 6000 mgL-1 had the highest number of lateral roots when planted from H2. Harvest 1 had greater quantities of lateral roots than H4, while both H1 and H4 were similar to H3. There was a harvest date x treatment interaction for total cutting dry weight. Main effects for evaluation week will be presented se parately. Differences in total cutting dry weight were not significant amongst treatmen ts for cuttings planted from H2 or H4 (Table 4-19). For cuttings planted from H1, those treated with Dip n’ Grow or KIBA at

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128 1500 mgL-1 were lighter than the other treat ments, while KIBA at 3000 to 6000 mgL-1 were similar to the control. Cuttings planted from H3 had greater dry weight when treated with concentrations of KIBA, as compared to the control, while Dip n’ Grow at 1500 mgL-1 was similar to all treatments. Table 4-18. Harvest date (H) by treatment in teraction for number of lateral roots of oleander cuttings planted duri ng the warm season (Expt. 1). Number of lateral roots Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 10.3 22.9 9.9 4.1 NS Dip n' Grow 1500 34.4 25.5 8.5 9.1 NS KIBA 1500 32.6 18.2 8.0 0.0 ** 13.5 KIBA 3000 21.6 33.2 11.4 10.0 16.8 KIBA 6000 20.8 50.0 6.7 1.7 ** 15.6 Significance * NS NS LSD 16.1 3.3 NS, *, ** Nonsignificant or significant at P 0.05 or 0.01, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt When considering only untreated cuttings, those planted from H4 were similar to H2 and generally lower in total dry weight than H1 and H3. Harvest 2 was similar to H3. Harvest 1 cuttings were only similar in dry weig ht to H3 and were heavier than the rest. For cuttings treated with Dip n’ Grow at 1500 mgL-1 all harvest plantings resulted with similar weighted cuttings, except H3 had h eavier cuttings than H4. KIBA treatments produced the lowest total dry weight when cuttings were planted from H4, while the other harvest dates were similar. Harvest 3 was greater than H1 for KIBA at 1500 mgL-1 and H1 was greater in total dry weight than H2. Harvests 1, 2, and 3 were all similar when treated with KIBA at 3000 mgL-1. Amongst treatments, total cutting dry weight was 584 mg per cutting 6 WAP, which wa s 116% greater than 4 WAP (505 mg) (LSD=39, n=71).

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129 The harvest date x treatment interaction was not significant for cutting root: shoot ratio, therefore the main effects will be pres ented separately. When comparing harvests, cuttings planted from H4 had at least a 37% lo wer root : shoot ratio than H1 to H3 (data not shown). For cuttings ev aluated 4 WAP, the root : s hoot ratio was 44% less than 6 WAP (0.10 vs 0.18) (LSD=0.02, n=71). Amongst all harvests, cuttings treated with KIBA at 1500 or 3000 mgL-1 or Dip n’ Grow at 1500 mgL-1 had similar root : shoot ratios (0.15), which was at least 138% greate r than the control (0.11). Cuttings treated with KIBA at 6000 mgL-1 (0.13) were similar to all treatments (LSD=0.03, n=28). Table 4-19. Harvest date (H) by treatment in teraction for total cutting dry weight of oleander cuttings planted during the warm season (Expt. 1). Total cutting dry weight (mg) Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 653 479 534 354 *** 133 Dip n' Grow 1500 491 482 606 371 136 KIBA 1500 454 549 646 251 *** 132 KIBA 3000 685 584 653 349 *** 150 KIBA 6000 766 607 681 386 *** 139 Significance *** NS NS LSD 155 99 NS, *, *** Nonsignificant or significant at P 0.05 or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt Experiment 2 There was a harvest date x evaluation week interaction for bot h percent survival and rooting quality. There was also a harves t date x treatment interaction for rooting quality. For percent survival, differences in evaluation week we re not significant for cuttings planted from H1, H2, and H3, with survival rates of 99%, 100, and 98%, respectively. For H4, cuttings evaluated 6 WAP had a 98% percent survival, which was greater than 4 WAP (91%) (LSD=6, n=20). Differences in rooting quality were not

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130 significant between evaluation weeks for H1 (3.4). For H2, cuttings evaluated 4 WAP (3.7) had lower rooting quality than those evaluated 6 WAP (4.33) (LSD=0.3, n=20). For H3 and H4, cuttings evaluated 6 WAP had higher rooting quality values than those evaluated 4 WAP (data not shown). Cuttings planted from H1 and treate d with Dip n’ Grow at 1500 mgL-1 or KIBA at 6000 mgL-1 were similar to the control, while KIBA at 1500 or 3000 mgL-1 had the best rooting quality values (Table 4-20). Differences in rooti ng quality were not significant amongst treatments for H2 or H4. From H3 however, cuttings had a higher quality ranking when treated with KIBA at 1500 mgL-1 than the control or KIBA at 3000 mgL1. Dip n’ Grow at 1500 mgL-1 and KIBA at 6000 mgL-1 were similar in rooting quality to all treatments. When considering only untreated cuttings, a ll harvests produced cuttings with similar rooting quality values, except when planted from H2, which was higher than the rest. For all rooting hormone concentrations, rooting quality was similar from each harvest throughout the experiment. There was a harvest date x treatment interaction for both primary and lateral root number. Cuttings planted from H1 had a sim ilar number of primary roots when treated with Dip n’ Grow at 1500 mgL-1 or left untreated; both of these treatments expressed a 38 % lower number of primary roots than a ny concentration of KI BA (Table 4-21). Cuttings had 16.7, 8.1, and 4.3 primary roots per cutting from H2, H3, and H4, respectively, and were not significan tly different amongst treatments. Untreated cuttings planted from all harvests had similar quantities of primary roots except H2, which had more than H1, H3, and H 4. For cuttings treated with Dip n’ Grow at 1500 mgL-1, those planted from H2 had a greater number of primary roots than H4.

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131 Harvest 3 and H1 had a similar number of prim ary roots to all harvest dates. Cuttings treated with KIBA at 1500 mgL-1 were generally similar when planted from each harvest, although H2 had a greater number of primary roots than H4. When considering higher rates of KIBA, thos e cuttings treated with concentrations of 3000 mgL-1 had at least 272% more primary roots from H1 and H2 than either H3 or H4. Similarly, when treated with KIBA at 6000 mgL-1, number of primary roots tended to decrease as the experiment progressed into December. Table 4-20. Harvest date (H) by treatment in teraction for visual rooting quality (1-5; 1=poor, 5=best) of oleander cuttings planted during the cool season (Expt. 2). Rooting quality Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 3.0 4.3 3.3 2.9 ** 0.8 Dip n' Grow 1500 2.9 3.9 3.7 3.3 NS KIBA 1500 3.9 4.3 4.0 3.1 NS KIBA 3000 4.1 3.8 3.5 3.0 NS KIBA 6000 3.2 4.1 3.6 3.2 NS Significance *** NS NS LSD 0.5 0.5 NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt When considering the lateral root number, cuttings of oleander responded similarly to all treatments when planted from all harves ts except H3. This particular harvest had similar rooting with all hormone treatments except KIBA at 1500 mgL-1, which had a greater number of lateral roots than the re st (Table 4-22). When considering only untreated cuttings, H2 had the most lateral r oots per cutting, and while H3 and H4 were similar in quantity, both were at least 148% greater than H4. Similarly, when treated with Dip n’ Grow at 1500 mgL-1, cuttings planted from H2 produced more lateral roots than any other harvest. Harvest 3 produced more lateral roots than H4, yet both were

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132 similar to H1. Differences in number of lateral roots were no t significant amongst harvest dates when treated with KIBA at 1500 mgL-1. Harvests 1 and 2 produced more lateral roots than H4, while H3 was similar to all harvest dates when treated with KIBA at 3000 mgL-1. When treated with KIBA at 6000 mgL-1 H2 produced more lateral roots than H1, while later harvests produced similar and lower values than the rest. Table 4-21. Harvest date (H) by treatment in teraction for number of primary roots of oleander cuttings planted duri ng the cool season (Expt. 2). Number of primary roots Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 7.5 15.8 7.7 5.4 6.3 Dip n' Grow 1500 7.6 11.9 9.1 4.9 2.3 KIBA 1500 12.3 18.7 9.3 3.6 ** 6.9 KIBA 3000 16.3 16.7 6.0 3.0 *** 3.7 KIBA 6000 14.7 20.2 8.5 4.6 *** 5.3 Significance ** NS NS NS LSD 4.3 NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt The harvest date x treatment interacti on was not significant for total cutting dry weight; therefore the main effects will be presented. Total cutting dry weight was similar for all treatments at 604 mg per cutting. Fo r cuttings evaluated 6 WAP (671 mg), total cutting dry weight was 125% greater than 4 WAP (537 mg) (LSD=37, n=80). When considering harvest date, cuttings planted from H2 had the great est total cutting dry weight and as the experiment progressed, re ductions in dry weight occurred (data not shown). There was a harvest date x treatment and a harvest date x evaluation week interaction for the cutting root : shoot rati o. From H1, KIBA treated cuttings had higher root : shoot ratios when compared to the control or Dip n’ Grow at 1500 mgL-1 (Table 4-

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133 23). Cuttings planted from H2 had similar root : shoot ratios for thos e treated with Dip n’ Grow, KIBA at 1500 mgL-1, or left untreated. Cuttings treated with KIBA at 6000 mgL1 had similar root : shoot ratios to all treatments, except KIBA at 3000 mgL-1, which was at least 17 % lower,. For cuttings planted from H3, all concentrations of KIBA were similar and at least 129% greater than the co ntrol, however those treated with Dip n’ Grow at 1500 mgL-1 had similar root : shoot ratios to all treatments. The cutting root : shoot ratios were similar for all treatments when cuttings were planted from H4. Table 4-22. Harvest date (H) by treatment in teraction for number of lateral roots of oleander cuttings planted duri ng the cool season (Expt. 2). Lateral roots Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 17.8 54.4 19.5 1.2 *** 11.8 Dip n' Grow 1500 10.7 44.9 18.0 2.3 *** 14.4 KIBA 1500 37.7 40.1 39.1 3.9 NS KIBA 3000 33.7 33.6 14.7 3.1 20.9 KIBA 6000 25.1 34.8 13.8 5.6 ** 12.3 Significance NS NS NS LSD 17.7 NS, *, **, *** Nonsignificant or significant at P 0.05, 0.01, or 0.001, respectively. Dip n’ Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt When considering only the control, H2 ha d greater root : shoot ratios as compared to all other harvests. For cuttings treated with Dip n’ Grow at 1500 mgL-1, H2 was greater than H1 or H4, yet H3 was similar to all harvests. When treated with KIBA at 1500 mgL-1, cuttings had higher root : shoot ratios from H2 than H4, while H1 and H3 were similar to all harvests. KIBA at concentrations of 3000 mgL-1 produced cuttings with greater root : shoot rati os from H1 than H4, while the other harvest dates were similar. For cuttings treated with KIBA at 6000 mgL-1 those planted from H4 had a lower root : shoot ratio than H1 or H2, wh ile H3 was similar to all harvests. When

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134 comparing evaluation date, cuttings planted from H1 had greater root : shoot ratios 6 WAP than 4 WAP (0.15 vs 0.11) (LSD=0.02, n= 20). Similarly, cuttings evaluated 6 WAP had greater root : shoot ratios than 4 WAP for all other harvest dates, although as the experiment progressed differences between evaluation dates tended to increase (data not shown). Table 4-23. Harvest date (H) by treatment in teraction for root : shoot ratio (shoot dry weight / root dry weight) of oleander cuttings plan ted during the cool season (Expt. 2). Root : shoot ratio Treatment Concn (mgL-1) H1 H2 H3 H4 LSD Control 0 0.10 0.16 0.09 0.08 *** 0.03 Dip n' Grow 1500 0.11 0.15 0.12 0.09 0.04 KIBA 1500 0.15 0.16 0.13 0.11 NS KIBA 3000 0.15 0.13 0.12 0.09 0.04 KIBA 6000 0.14 0.14 0.12 0.09 NS Significance *** * NS LSD 0.02 0.02 0.03 NS, *, *** Nonsignificant or significant at P 0.05 or 0.001, respectively. Dip n’ Grow = 1% indolebutyric ac id : 0.5% napthaleneacetic acid KIBA = indolebutyric acid, potassium salt Discussion During the first experiment percent survival of cuttings was generally not enhanced by rooting hormone applications when co mpared to the control; however at H3, applications of all rooting hormone treatments were shown to have greater survival percentages than the untreated cuttings. Visual root quality ratings were generally greater for KIBA treatments as compared to the contro l, yet Dip n’ Grow was shown to be either similar or in lower quantity when compared to the control. Number of primary roots was not influenced by hormone treatments. App lication of rooting hormones improved lateral

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135 root number when cuttings were planted fr om H1 and H2. As the season progressed, lateral root number was less responsive to treatment. Total dry weight was genera lly not influenced by applic ation of rooting hormones, although from H3 all concentrations of KIBA had a greater total cutt ing dry weight than the control. The cutting root : shoot ratio was also enhanced with application of KIBA at 1500 to 3000 mgL-1 or Dip n’ Grow at 1500 mgL-1. During the second experiment, percent surv ival was similar for all treatments. Supplemental applications of KIBA at concentrations of 1500 to 3000 mgL-1 were generally shown to improve visual rooting quality of cuttings planted from H2 and H4, otherwise all treatments had similar results. Cuttings treated with KI BA tended to have a greater number of primary roots as compared to Dip n’ Grow or the control, although most harvests showed similarities amongst trea tments with respect to number of primary roots. Similarly, lateral roots were enhanced by app lication of KIBA at 1500 mgL-1 from H3, and generally not influenced from other harvests. Total cutting dry weight was similar for all treatments, yet the root : shoot ratio was generally greater when cuttings were treated with KIBA. When considering both experiments, ther e was an overall decline in quality of oleander cuttings as the durati on of the studies progressed. Th is could be attributed to many physiological factors such as season, nutrient content, or deficiencies in carbohydrate accumulation do to aggressive harvesting of stock plants. Although the two experiments were not compared, both suggest a similar trend toward reduction of quality at later harvests, therefore the effect does not appear to be seasonal.

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136 At times application of Dip n’ Grow at 1500 mgL-1 produced lower rooting values than the control and this may suggest sensit ivity of oleander cuttings to alcohol-based rooting hormone formulations. Conclusion Evidence was provided in support of enhanced propagation of oleander through application of KIBA at con centrations of 1500 to 3000 mgL-1 during both seasons. Currently, KIBA has limited commercial av ailability with a cost of $81.60 per 25 g (Sigma-Aldrich Co., St. Lois, Mo.), while Di p n’ Grow is a commercially available product with a cost of $190.00 per 3.8 L (M orton’s Horticultural Products Inc., McMinnville, Tenn.). Application of Di p n’ Grow or KIBA mixed at 1500 mgL-1 would cost a propagator $5.02 and $4.90 per L (exclu ding labor), respectively. Propagators should consider cost of application and benef its achieved in rooting performance before deciding on a rooting hormone program.

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137 Table 4-1. Temperature and relative humidity data as recorded during simulated and actual shipping (excision to planting) of cuttings from Harvests (H) 1, 2, 3, and 4 of Expt. 1 and 2, respectively. Expt 1 Temperature (C) Relative Humidity (%) Harvest Date Max Min Avg Max Min Avg H1 6 June 33 11 20 95 48 81 H2 27 June 31 14 23 96 49 80 H3 18 July 34 20 26 94 57 78 H4 8 Aug. 43 20 26 86 43 67 Expt 2 Temperature Relative Humidity Harvest Date Max Min Avg Max Min Avg H1 6 Sept. 34 2 24 85 31 65 H2 27 Sept. 32 3 19 93 32 73 H3 18 Oct. 31 3 21 92 31 69 H4 8 Nov. 33 3 17 80 21 46

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138 CHAPTER 5 RESEARCH SUMMARY A series of greenhouse-based experiment s were conducted during the 2005 summer (warm season) and 2005-2006 fall/winter (cool se ason) to investigate the influence of nutrition, chemical branching ag ents, or rooting hormone tr eatments on stock plant yield of tropical perennials and subsequent root ing performance harvested of cuttings. In the first set of experiments stock plants of Bougainvillea glabra (Choisy.) 'Raspberry Ice' and 'Purple Small Leaf' were fertilized with three concentrations of nitrogen (N) at 100, 200, or 300 mgL-1 to determine optimum N fertigation standards for maximizing yield of high quality stem cuttings that root the best. Of the concentrations studied, the range of 200 to 300 mgL-1 was shown to effectively increase cutting yield and improve subsequent rooting performance. Concentrations s hould be provided at 300 mgL-1 during the active growth periods associated with summer, and reduced to 200 mgL-1 as daylength becomes shorter, temperatures decrease, and light quantity is reduced during fall/winter months. In order for growers to profit from the sp ecialized handling of tropical perennials, production protocols must be establishe d. One step towards establishing a comprehensive stock plant management progr am was defining the optimal nitrogen (N) fertilization rates for maximum cutting quant ity and high rooting performance. By implementing these recommendations growers can be better prepared to meet the increased market demands for tropical perennial s. Further research should be conducted

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139 to determine if the recommended N concentr ation range is appropriate application on other tropical species. In a second set of experiments, stock plants of Tecoma stans (L.) Juss. ‘Esperanza’ and Nerium oleander (L.) ‘Dwarf Salmon’ were spra yed with commercially available lateral branching agents to identify appr opriate treatments for increasing yield and enhancing harvested cutting rooting performance. Results indicated that concentrations greater than 125 mgL-1 may have been excessive, as phytotoxic symptoms were shown to damage or distort cutting tissue and lim it rooting. Fresco, a lthough highly phytotoxic to both species, tended to increase cutti ng yield in oleander. FAL-457 was shown to increase yield, however rooting was hindered. Florel was shown to increase yield with similar rooting to the control, unfortunately cutting stunting and tipburn occurred at the concentrations applied. Alt hough the lateral branching treatments applied did not prove to be beneficial at the concen trations investigated, the tre nds in Chapter 3 coupled with other background research on traditional floric ulture crops support a need for future research on oleander and tecoma and other tr opical species. Some areas for further investigation may include synergistic tank mixes to create desired attributes in both stock plant and liner tray cuttings. One example of employing strengths of one ingredient to compensate for weaknesses of another may be the combination of Florel (ability to increase yield and subsequent rooting) and Fresco (stem elongation) to produce optimally sized liners. There is an increasing demand for cuttings of tropical perennials to root more uniformly and in less time under mist. In a th ird series of experiments several species of tropical perennials were treat ed with commercially-available rooting hormones to

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140 measure the impact on rooting performance. Treatments consisted of an untreated control, a water-soluble rooting hormone i ndole-3-butyric acid pot assium salt (KIBA) applied at concentra tions of 1500 to 6000 mgL-1, or the standard alcohol-based Dip N’ Grow at 1500 mgL-1. Results indicated KIBA was an effective formulation for propagation of alcohol-sensitive species such as oleander, an d was comparable to Dip N’ Grow. Although recommended concentrations vary with species investigated, a KIBA range of 3000 to 6000 mgL-1 was shown to be most effective for increasing cutting rooting performance. Currently, KIBA has limite d commercial availability with a cost of $81.60 per 25 g (Sigma-Aldrich Co., St. Lois, M o.), while Dip n’ Grow is a commercially available product with a cost of $190.00 per 3.8 L (Morton’s Horticultural Products Inc., McMinnville, Tenn.). Application of Di p n’ Grow or KIBA mixed at 1500 mgL-1 would cost a propagator $5.02 and $4.90 per L (exc luding labor), respectively. Before propagators decide on a rooting hormone treatment they should consider both the cost of a rooting hormone application and the benefits achieved in propagati on. Further research should investigate optimum rooting hormone concentrations for propagation of other tropical species.

PAGE 155

141 LIST OF REFERENCES Aldrich, J.H. and J.G. Norcini. 1995. Coppe r hydroxide-treated pots improve the root system of Bougainvillea cuttings. Proc Annu. Mtg. Fla. State Hort. Soc. 107:215217. Arent, G.L. and M.L. Voigt. 1994. Lansdcaping, p. 75-91. In: E.J. Holcomb (ed.). Bedding plants IV : a manual on the culture of bedding plants as a greenhouse crop. Ball Publishing, Batavia, Ill. Arias, S.B., J.A.F. Leemhuis, J.A.F. Herna ndez, J.O. Rego, and A.G. Benavente-Garcia. 2001. Growth and leaf colour responses of oleander (Nerium oleander L.) to pinching and chlormequat chloride treatment. Acta Hort. 559:155-160. Atzmon, N., Z. Wiesman, and P. Fine. 1997. Bi osolids improve rooting of bougainvillea (Bougainvillea glabra) cuttings. J. Environ. Hort. 15(1):1-5. Auld, R.E. 1987. The basics of propagating Bougainvillea. Proc. Intl. Plant Prop. Soc. 36:211-213. Ball, V. 1998. Ball Redbook. 16th ed.. Ball Pub., Batavia, Ill. Bell, M.L., R.A. Larson, and D.A. Bailey. 1997. Vegetative growth responses of florist azaleas to dikegulac, GA4+7, and benzylamino purine. HortScience 32(4):690-693. Bhat, N.R., D.S. Baraskar, and K.G. Choudha ri. 1988. Studies on roo ting of difficult-toroot bougainvillea varieties. HortScience 23(3):752-752. Bhattacharjee, S.K. and M.B. Balakrishna. 1983. Propagation of Bougainvillea from stem cuttings. II. Effect of growth regulator s. Haryana J. Hort. Sci. 12(1 /2):13-18. Blazich, F.A. 1988a. Chemical formulations used to promote adventitious rooting, p. 132-149. In: T.D. Davis, B.E. Haissig, and N. Sankhla (eds.). Adventitious root formation in cuttings. Diosco rides Press, Portland, Ore. Blazich, F.A. 1988b. Mineral nutrition and adven titious rooting, p. 61-69. In: T.D. Davis, B.E. Haissig, and N. Sankhla (eds.). Adve ntitious root formation in cuttings. Dioscorides Press, Portland, Ore. Bowden, R. 2004. Growing tropicals out of zone. Ornamental Outlook 13(2):14,16

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142 Cerveny, C.B., J.L. Gibson, and J.E. Barrett. In press. Effects of three nitrogen fertilizer concentrations on stock plant production of Bougainvillea 'Raspberry Ice' and 'Purple Small Leaf'. Proc. Intl. Plant Prop. Soc. Chakraverty, R.K. 1970. Propagation of Bougainvillea by stem cuttings. Cur Sci. 39(20):472. Courtright, G. 1988. Tropicals. Ti mber Press, Portland, Ore. Czekalski, M.L. 1989. The influence of auxins on the rooting of cuttings of Bougainvillea glabra Choisy. Acta Hort. (251):345-352. Davidson, H., R. Mecklenburg, and C. Peterson. 2000. Nursery management : administration and culture. 4th ed. Pr entice Hall, Upper Saddle River, NJ. Davis, T.D., B.E. Haissig, and N. Sankhla. 1988. Adventitious root formation in cuttings. Dioscorides Press, Portland, Or. Dole, J.M. and H.F. Wilkins. 2005. Floricultu re : principles and species. 2nd ed. Prentice Hall, Upper Saddle River, N.J. Dress, W.J. 1974. Notes on a mandevilla (d ipladenia) hybrid (A pocynaceae). Baileya 19(3):106-110. Florida Nursery, Growers and Landscape Asso ciation. 2005. Plants of the Year Program Markets Proven Florida Plants. 6 April 2006. . Faust, J.E. and K.P. Lewis. 2005. The effects of Ethephon on cutting yield of 23 selected annual cultivars. Acta Hort. 683:141-144. Gibson, J.L. and B.E. Whipker. 2004. Ethephon and timing of Scaevola aemula stock plants influence vegetative cutting quanti ty and quality. Plant Growth Regulat. Soc. Amer. Qrtly. 32(4):119-123. Ganmore-Neuman, R.A.A.H. 1990. Effect of the NO3-/NH4+ ra tio in nutrient solution on Pelargonium stock plants: yield and quali ty of cuttings. J. Plant Nutr. 13:12411256. Ganmore-Neuman, R.A.A.H. 1992. Plant growth and cuttin g production of containergrown Pelargonium stock plants as aff ected by N concentration and N form. J. Amer. Soc. Hort. Sci. 117:234-238. Griffiths, M. 1994. Index of garden pl ants. Timber Press, Portland, Ore. Hackett, W.P., R.M. Sachs, and J. Debi e. 1972. Growing Bougainvillea as a flowering pot plant. Calif. Agr. 26(8):12-13.

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143 Hamrick, D., 2003. Introduction, p. 3-8. In: D. Hamrick Ed., Ball Redbook Vol. 2. 17th ed. Ball Publishing, Batavia, Ill. Hartmann, H.T., D.E. Kester, Fred Davies, and R. Geneve. 2002. Hartmann and Kester's plant propagation : principles and practic es. 7th ed. Prentice Hall, Upper Saddle River, N.J. Howard, B.H. 1994. Manipulating rooting poten tial in stock plants before collecting cuttings., p. 123-142. In: T.D. Davis a nd B.E. Haissig (eds.). Biology of adventitious root formati on. Plenum Press, New York. Judd, W.S., C. S. Campbell, E. A. Ke llogg, P. F. Stevens, and M.J. Donoghue. 2002. Plant systematics : a phylogenetic ap proach. 2nd ed. Sinauer Associates, Sunderland, Mass. Konjoian, P. 1994. Florel treatment of stoc k plants; Where are we heading?. KFES Newsnotes. 94-8. McAvoy, R. 1995. Basic principles of vege tative plant propagation. Conn. Greenhouse Nwsltr:1-6. McCulla, F.A.C. 1973. Oleanders. [Nerium olea nder, culture]. Horticulture 50(7):26-27 Mudge, K.W., V.N. Mwaja, F.M. Itulya, a nd J. Ochieng. 1995. Comparison of 4 moisture management-systems for cutting propaga tion of bougainvillea, hibiscus, and kei apple. J. Amer. Soc. Hort. Sci. 120(3):366-373. Mynett, K. 1985. Growing tree-like plants of fuschia by using gibberellic acid/ GA3. Acta Hort. 167:333-338. National Agricultural Statis tics Service, USDA. 2005. Flor iculture Crop Statistics. 9/26/2005 . Nau, J. 1999. Ball culture guide : the encycl opedia of seed germination. 3rd ed. Ball Publishing, Batavia, IL. Palmer, D. 2000. The Retail Gardener. 21 May 2006. <>. Popenoe, J. 1976. Oleander cultivars at the Fa irchild Tropical Garden. Proc. Annu. Mtg. Fla. State Hort. Soc. 88:426-428. Puglisi, S.E. 2002. Use of plant growth regula tors to increase branching of Clematis Spp. Va. Poly. and State Univ., Blacksburg, MS Thesis. Rama Rao, B.A. 1976. A handbook on bougainvillea culture. B.A. Rama Rao Publishing, New Delhi.

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144 Roeber, R. and G. Reuther. 1982. Der ei nfluss unterschiedlicher N-formen undkonzentrationen auf den ertrag und die qua litat von chrysanthermen-steklingen. Gartenbauwissenschaft 47:182-188. Schoellhorn, R. and E. Alvarez, 2002. Warm climate production guidelines for Bougainvillea Vol. ENH874, EDIS, University of Florida, pp. 5. Singh, B., P.K. Majumdar, and J. Prasad. 1976. Response of bougainvillea cuttings under bottom heat. Delhi Garden Mag. 13: p 13. Srivastava, L.M. 2002. Plant growth and de velopment : hormones and environment. Academic Press, Amsterdam. Walker, J. and G. Hanly. 1996. The subtropical garden. Timber Press, Portland, Ore. Whipker, B.E., T.J. Cavins, J.L. Gibson, J.M. Dole, P.V. Nelson, and W. Fonteno, 2003. Plant nutrition, p. 29-37. In: D. Hamrick Ed., Ball Redbook Vol. 2 17th ed., Ball Publishing, Batavia. Witaszek, W.A.K.M. 1989. Light and growth regulators in the cuttings production of poinsettia/ Euphorbia pulcherrima/ mo ther plants. Acta Hort. 251:315-317. Wolnik, D.J. 1994. Merchandising bedding pl ants, p. 51-60. In: E.J. Holcomb (ed.). Bedding plants IV : a manual on the culture of bedding plants as a greenhouse crop. Ball Publishing, Batavia, Ill. Yadav, L.P., A.P. Bhattacharya, and H.S. Pandey. 1978. Effect of season on the rooting of Bougainvillea cuttings. Prog. Hort. 9(4):72-73.

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145 BIOGRAPHICAL SKETCH Christopher Cerveny earned his Bachelor of Science degree in horticulture from Michigan State University in 2002. His flor iculture industry experience includes 2 years as propagation manager for Kalamazoo Speci alty Plants, Kalamazoo, MI. Currently, Chris is a graduate research assistant at the University of Florida where his contributions to the art and science of horti culture continue to increase. Chris has published several articles in trade journals, magazines, and c onference proceedings. He has been invited to speak at a number of horticulture meetings and was a contributing author to Dole and Gibson’s 2006 text, Cutting Propagation: A Guide to Propagating and Producing Floriculture Crops Chris is an active member in the Environmental Horticulture Graduate Student Association at the Univ ersity of Florida and has a tremendous appreciation for all who are passionate about plants.


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Table of Contents
    Title Page
        Page i
        Page ii
    Dedication
        Page iii
    Acknowledgement
        Page iv
    Table of Contents
        Page v
        Page vi
        Page vii
    List of Tables
        Page viii
        Page ix
        Page x
        Page xi
    List of Figures
        Page xii
    Abstract
        Page xiii
        Page xiv
    Literature review
        Page 1
        Page 2
        Page 3
        Page 4
        Page 5
        Page 6
        Page 7
        Page 8
        Page 9
        Page 10
        Page 11
        Page 12
        Page 13
    Stock plant nutrition
        Page 14
        Page 15
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    Lateral branching agents
        Page 55
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    Rooting hormones
        Page 81
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    Research summary
        Page 138
        Page 139
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    References
        Page 141
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    Biographical sketch
        Page 145
Full Text












STOCK PLANT MANAGEMENT OF TROPICAL PERENNIALS


By

CHRISTOPHER BRIAN CERVENY













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


2006

































Copyright 2006

by

Christopher Brian Cerveny

































This document is dedicated with expressed appreciation to the "Research Machine"
Cricket, Dawn, Jude, Leah, and Neil.















ACKNOWLEDGMENTS

I thank my graduate committee, Drs. James E. Barrett, Brian E. Myers, and James

L. Gibson, for their guidance and support throughout the degree process. I thank Carolyn

Bartuska and Dr. Grady L. Miller for statistical assistance. For numerous hours of

assistance during experiment preparations and collection of data, I wish to thank Jeffrey

Anderson, Shannon Crowley, Neil Greishaw, Jude Groninger, Alina Lovelace, Leah

McCue, Meghan Pressley, Dawn Taylor, Carson Winn, and Mariah Williams. Additional

appreciation is expressed to Bob Weidman for assistance with growth regulator

applications and for providing student labor at even the shortest of notice. I thank the

Fred C. Gloeckner Foundation for grant support and Hatchett Creek Farms for donation

of plant materials. For chemical donations throughout the experiment process

appreciation is given to Fine Agrochemicals, Monterey Garden Supply, and Dip n' Grow

Inc. Lastly, I wish to thank my parents for expressing pride in my accomplishments.

This was a source of motivational support that is without comparison.
















TABLE OF CONTENTS



A C K N O W L E D G M E N T S ................................................................................................. iv

LIST OF TABLES .......................................... viii

LIST OF FIGURES ............................. .. .......... .................................. xii

ABSTRACT ........................................ .................................. xiii

CHAPTER

1 LITERATURE REVIEW ...............................................................................1......

Introduction to the Floriculture Industry ................................................................1...
A sexual Propagation of C uttings ..................................... ..................... ...............3...
Physiological Age of Cuttings......................................................................3...
G row ing E nvironm ent .......................................... ......................... ...............4...
Mineral Nutrition ............................ ................................................................... 5
Growth Regulator Treatments ......................................................................6...
Rooting Hormone Treatments ......................................................................7...
C o n c lu sio n ...................................................................................................... 8
Selected Tropical Species .............. .... ............. ................................................ 9
A llam anda schottii ..... .................................................................. .................
B ougainvillea g labra .......................................... ......................... ................ 10
M andevilla sp lendens ....................... ......................................................1...... 1
Nerium oleander ....................................... .......................... ...... ............. 12
T eco m a sta n s ....................................................................................................... 13
O b j e c tiv e s ...................................................................................................................1 3

2 STOCK PLANT NUTRITION.............................................................................. 14

In tro d u c tio n ................................................................................................................. 1 4
M materials and M ethods .. ..................................................................... ............... 15
E x p erim en t 1 ....................................................................................................... 15
E x p erim en t 2 ....................................................................................................... 19
R e su lts....................................................................................................... ....... .. 19
E x p erim en t 1 ....................................................................................................... 19
Stock Plant Parameters ............................................................................ 19
Cutting quantity ............ .......................................... 19


v









Cutting length, stem diameter, and leaf area...........................................20
Rooting Parameters ................ ....... .... .................... ............... 21
Percent survival, visual rooting quality, and quantity of primary and
lateral roots ...... ............. .. ............................................ 21
Cutting dry weight and root : shoot ratio ................................................24
Stock plant leachate...................................... .. .......... .......... ................. 26
F oliar analy sis ..................................................................................... 27
E x p erim en t 2 ....................................................................................................... 2 8
Stock plant param eters ........................................ ........................ ................ 28
C cutting quantity .......................................... ............ ...... ... ........ ..... 28
Cutting length, stem diameter, and leaf area...........................................29
R ooting param eters..................................................... ........ ............ ................ 1
Percent survival, visual rooting quality, and quantity of primary and
lateral roots .............. . ............................................. 3 1
Cutting dry weight and root : shoot ratio ................................................33
Stock plant leachate...................................... .. .......... .......... ................. 34
F oliar analy sis ..................................................................................... 34
D isc u ssio n ............................................................................................................... ... 3 6
C o n c lu sio n ............................................................................................................... .. 4 1

3 LATERAL BRANCHING AGENTS ....................................................................55

In tro d u ctio n ............................................................................................................... .. 5 5
M materials and M ethods .. ..................................................................... ................ 56
E x p erim en t 1 ....................................................................................................... 5 6
E x p erim en t 2 ....................................................................................................... 5 7
Harvesting procedure ........................................................57
C cutting evaluations ..................................... .. .......... ............ .. ........ .... 58
R e su lts....................................................................................................... ....... .. 5 8
E xperim ent 1 .................... .... .. ... ... .. ............................... 58
Harvest yield, cutting length, and stem diameter....................................58
Percent survival and visual rooting quality .............................................62
Cutting total dry weight and root : shoot ratio ........................................63
E xperim ent 2 ..... .............. .... ...... ... .. ............................... 65
Harvest yield, cutting length, and stem diameter ....................................65
Percent survival and visual rooting quality .............................................69
Cutting total dry weight and root : shoot ratio ........................................71
D isc u ssio n ............................................................................................................... ... 7 4
C o n c lu sio n ................................................................................................................. 7 9

4 R O O TIN G H O R M O N E S .......................................... ......................... .................. 81

In tro d u c tio n ................................................................................................................. 8 1
M materials and M ethods .. ..................................................................... ................ 82
E x p erim en t 1 ....................................................................................................... 8 2
Harvesting procedure ........................................................83
C cutting evaluations ..................................... .. .......... ............ .. ........ .... 83









E xperim ent 2 ................................................................................................. 84
R e su lts...........................................................................................................8 5
A llam anda schottii .........................................................................................85
Experiment 1 ............................ ............................ 85
E xperim ent 2 ..............................................................................................9 1
D discussion ............................................................................................... 96
C conclusion ........................................................... ..........................98
Bougainvillea glabra 'California Gold' ...........................................................98
E xperim ent 1 ..............................................................................................98
E x p erim en t 2 ......................................................................... ...............10 3
Discussion .......................... ..............................105
Conclusion........................... ............................. ............... 107
Bougainvillea glabra 'Helen Johnson'...... ... .......................................107
E x p erim en t 1 ......................................................................... ...............10 7
E x p erim en t 2 ......................................................................... ...............1 12
Discussion .......................... ..............................118
C conclusion ..................................................................................................119
Mandevilla splendens 'White'.......................................120
E x p erim en t 1 ......................................................................... ...............12 0
E xperim ent 2 .............. .................. ............................................... 12 1
Discussion .......................... ..............................123
C conclusion ............................................................................................124
Nerium oleander 'Dwarf Salmon'. .. ............ ....................... ................ 125
E x p erim en t 1 ......................................................................... ...............12 5
E xperim ent 2 .............. .................. .............................................. 129
Discussion .......................... ..............................134
C onclu sion ............................................................................................136

5 RESEAR CH SUM M ARY ............................... ........................ ................ 138

L IST O F R E FE R E N C E S ......................................................................................14 1

BIOGRAPHICAL SKETCH .......................... .............. ............................................... 145















LIST OF TABLES


Table page

2-1 Temperature and relative humidity data (maximum, minimum, and mean) as
recorded during simulated and actual shipping...................................................42

2-2 Influence of harvest date, cultivar, and nitrogen (N) concentration on quantity of
sub-term inal bougainvillea stem cuttings .......................................... ................ 42

2-3 Significance levels for effects of harvest date, cultivar, and nitrogen (N)
concentration on length and leaf area of cuttings................................................43

2-4 Harvest date (H) by nitrogen (N) concentration interaction for length of sub-
term inal bougainvillea stem cuttings................................................... ................ 43

2-5 Significance levels for effects of harvest date, evaluation date, cultivar, and
nitrogen concentration .................................................................................. 44

2-6 Harvest date (H) by nitrogen (N) concentration interaction for percent survival
of sub-term inal bougainvillea.............................................................. ................ 44

2-7 Harvest date (H) by cultivar by N concentration interaction for visual rooting
quality (l=poor; 5=best) ...................................................................... 45

2-8 Significance levels for effects of harvest date, cultivar, and nitrogen
co n cen tratio n ............................................................................................................ 4 5

2-9 Harvest date by cultivar by N concentration interaction for total cutting dry
w eight (shoot w eight + root w eight) ................................................... ................ 45

2-10 Significance levels for effects of harvest date, cultivar, and nitrogen (N)
concentration on foliar concentration.................................................. ................ 46

2-11 Harvest date (H) by cultivar interaction for foliar nutrient concentration of
m acro n u trien ts .......................................................................................................... 4 6

2-12 Harvest date (H) by nitrogen (N) concentration interaction for foliar nutrient
concentrations of m acronutrients ........................................................ ................ 47

2-13 Cultivar by nitrogen (N) concentration interaction for foliar nutrient
concentration of m acronutrients.......................................................... ................ 47









2-14 Influence of harvest date, cultivar, and nitrogen (N) concentration on quantity of
sub-term inal bougainvillea stem cuttings........................................... ................ 48

2-15 Significance levels for effects of harvest date, cultivar, and nitrogen (N)
co n cen tratio n ............................................................................................................ 4 8

2-16 Cultivar by nitrogen (N) concentration interaction for leaf area of sub-terminal
bougainvillea stem cuttings harvested during the cool season (Expt. 2)...............49

2-17 Significance levels for effects of harvest date, evaluation date, cultivar, and
nitrogen concentration ...................................................................... ............ 49

2-18 Harvest date (H) by cultivar by nitrogen (N) concentration interaction for visual
ro o tin g q u ality .......................................................................................................... 5 0

2-19 Influence of harvest date (H), cultivar, and nitrogen (N) concentration on
quantity of prim ary roots......................................... ......................... ................ 50

2-20 Significance levels for effects of harvest date, cultivar, and nitrogen (N)
concentration on foliar concentration of macronutrients ....................................51

2-21 Harvest date (H) by cultivar by nitrogen (N) concentration interaction for foliar
nutrient concentration ... .............................................................. 51

3-1 Significance levels for effects of harvest date, cultivar, and branching treatment
on quantity, length, and stem diam eter .............................................. ................ 59

3-2 Cultivar by treatment interaction for quantity and length of 2-node stem cuttings .59

3-3 Significance levels for effects of harvest date, cultivar, and lateral branching
treatment on percent survival, visual rooting quality ..........................................63

3-4 Harvest date by cultivar by branching treatment interaction for rooting quality
(l=poor; 5=best) and percent survival ................................................ ................ 64

3-5 Significance levels for effects of harvest date, cultivar, and branching treatment
on quantity, length, and stem diameter of cuttings harvested from stock plants of
oleander and tecoma during the cool season (Expt. 1)........................................65

3-6 Influence of lateral branching agents on number and length of cuttings and
subsequent rooting perform ance ......................................................... ................ 66

3-7 Influence of lateral branching agents on quantity and length of cuttings and
sub sequent dry w eights .......................................... .......................... ................ 67

3-8 Influence of lateral branching agents on quantity and length, and subsequent
ro o tin g .................................................................................................................. ... 7 0









3-9 Influence of lateral branching agents on number and length of cuttings, and
subsequent rooting perform ance ......................................................... ................ 71

3-10 Significance levels for effects of harvest date, cultivar, and lateral branching
treatment on percent survival, visual rooting quality... ...................... ................ 80

4-3 Harvest date (H) by treatment interaction for visual rooting quality (1-5; l=poor,
5=best) of allamanda cuttings planted during the warm season (Expt. 1). .............86

4-4 Harvest date by treatment interaction for number of primary roots of stem
cuttings of allamanda planted during the warm season (Expt 1). ............................88

4-5 Harvest date (H) by treatment interaction for number of lateral roots of
allamanda cuttings planted during the warm season (Expt. 1) .................................88

4-6 Harvest date by treatment interaction for total cutting dry weight of allamanda
cuttings planted during the warm season (Expt. 1) ............................ ................ 91

4-7 Harvest date by treatment interaction for the cutting root : shoot ratio of
allamanda cuttings planted during the warm season (Expt. 1)...............................91

4-9 Harvest date by treatment interaction for number of primary roots of allamanda
cuttings planted during the cool season (Expt. 2). .............................. ................ 94

4-2 Influence of rooting hormone application on percent survival of allamanda
cuttings evaluated ................. .. ........... ........................................99

4-8 Influence of rooting hormone application on the visual rooting quality of
allam anda cuttings evaluated ........................................................ 100

4-10 Influence of rooting hormone on cutting root : shoot ratio of allamanda cuttings
ev a lu a te d ............................................................................................................ .. 1 0 1

4-11 Harvest date (H) by treatment interaction for bougainvillea 'California Gold'
cuttings planted during the warm season (Expt. 1). .................... ................... 104

4-12 Harvest date (H) by treatment interaction for root : shoot ratio (root weight +
shoot weight) of 'Helen Johnson' bougainvillea...... ................... .................. 110

4-13 Harvest date (H) by treatment interaction for visual rooting quality (1-5; l=poor,
5=best) of bougainvillea...................................... ........................ ............... 116

4-14 Harvest date (H) by treatment interaction for number of primary roots of
bougainvillea 'Helen Johnson' cuttings planted during the cool season (Expt. 2). 116

4-15 Harvest date (H) by treatment interaction for number of lateral roots of
bougainvillea 'Helen Johnson' cuttings planted during the cool season (Expt. 2). 117









4-16 Harvest date by treatment interaction for visual rooting quality (1-5; l=poor,
5=best) of mandevilla cuttings planted during the cool season (Expt. 2). ............122

4-17 Harvest date (H) by treatment interaction for visual rooting quality (1-5; l=poor,
5=best) of oleander cuttings planted during the warm season (Expt. 1). ..............126

4-18 Harvest date (H) by treatment interaction for number of lateral roots of oleander
cuttings planted during the warm season (Expt. 1) ...................... ................128

4-19 Harvest date (H) by treatment interaction for total cutting dry weight of oleander
cuttings planted during the warm season (Expt. 1) ...................... ................129

4-20 Harvest date (H) by treatment interaction for visual rooting quality (1-5; l=poor,
5=best) of oleander cuttings planted during the cool season (Expt. 2) ............... 131

4-21 Harvest date (H) by treatment interaction for number of primary roots of
oleander cuttings planted during the cool season (Expt. 2)...............................132

4-22 Harvest date (H) by treatment interaction for number of lateral roots of oleander
cuttings planted during the cool season (Expt. 2). .................................... 133

4-23 Harvest date (H) by treatment interaction for root : shoot ratio (shoot dry weight
/ root dry weight) of oleander cuttings planted during the cool season (Expt. 2). .134

4-1 Temperature and relative humidity data as recorded during simulated and actual
sh ip p in g ............................................................................................................. . 1 3 7















LIST OF FIGURES


Figure page

2-1 Nitrogen (N) concentration x sampling date interaction for leachate pH collected
from bougainvillea stock plants fertilized with................................... ................ 52

2-2 Main effects for leachate electrical conductivity (EC) over time, as recorded
from bougainvillea stock plants during the warm season (Expt. 1). .....................52

2-3 Nitrogen (N) concentration x sampling date interaction for nitrate nitrogen
(NO3-N) values recorded from stock plant leachate ........................... ................ 53

2-4 Nitrogen (N) concentration by sampling date interaction for leachate pH
collected from bougainvillea stock plants .......................................... ................ 53

2-5 Main effects for leachate electrical conductivity (EC) over time as recorded
from bougainvillea stock plants during the......................................... ................ 54

2-6 Nitrogen (N) concentration by sampling date interaction for leachate nitrate
nitrogen (NO3-N) values recorded from stock plants..........................................54















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

STOCK PLANT MANAGEMENT OF TROPICAL PERENNIALS

By

Christopher Brian Cerveny

August 2006

Chair: James L. Gibson
Major Department: Environmental Horticulture

A series of greenhouse-based propagation experiments were conducted during the

2005 summer (warm season) and 2005-2006 fall/winter (cool season) to investigate the

influence of nutrition, chemical branching agents, or rooting hormone treatments on the

stock plant yield of tropical perennials and subsequent rooting performance of cuttings.

In the first set of experiments stock plants of Bougainvillea glabra (Choisy.) 'Raspberry

Ice' and 'Purple Small Leaf were fertilized with three concentrations of nitrogen (N) at

100, 200, and 300 mg-L-1 to determine optimum N fertigation standards for maximizing

yield of high quality stem cuttings that root the best. Of the concentrations studied, the

range of 200 to 300 mg-L-1 was shown to effectively increase cutting yield and improve

subsequent rooting performance. Concentrations should be provided at 300 mg-L-1

during the active growth periods associated with summer, and reduced to 200 mg-L-1 as

daylength becomes shorter, temperatures decrease, and light quantity is reduced during

fall/winter months. In a second set of experiments, stock plants of Tecoma stans (L.)

Juss. 'Esperanza' and Nerium oleander (L.) 'Dwarf Salmon' were sprayed with









commercially available lateral branching agents to identify appropriate treatments for

increasing yield and enhancing rooting performance. Results indicated that

concentrations applied may have been excessive, as phytotoxic symptoms were shown to

damage or distort cutting tissue and limit rooting. Fresco, although highly phytotoxic to

both species, tended to increase cutting yield in oleander. FAL-457 was shown to

increase yield; however rooting was hindered. Florel was shown to increase yield with

similar rooting to the control; unfortunately cutting stunting and tip-burn occurred at the

concentrations applied. Further investigation with these species should be conducted at

lower concentrations or with combination lateral branching agent mixtures. In a third

series of experiments several species of tropical perennials were treated with rooting

hormones to measure the effect of synthetic auxin on rooting. Treatments consisted of an

untreated control, a water-soluble rooting hormone indole-3-butyric acid, potassium salt

(KIBA) applied at concentrations of 1500 to 6000 mg-L-1, or the standard alcohol-based

Dip N' Grow at 1500 mg-L1. Results indicated KIBA was an effective formulation for

propagation of alcohol-sensitive species such as oleander, and was comparable to Dip N'

Grow. Although concentrations varied with species investigated, a recommended KIBA

range of 3000 to 6000 mg-L1 was shown to be most effective for increasing cutting

rooting performance.














CHAPTER 1
LITERATURE REVIEW

Introduction to the Floriculture Industry

The U.S. Floriculture Industry has changed in several ways over the past 50 years.

From garden plants to house plants and cut flowers, florists no longer are the primary

distributor for horticultural products. Presently, floriculture crops are marketed through a

wide variety of sales outlets, from retail greenhouses and garden centers, to supermarkets,

large chain stores, and seasonal road-side stands (Ball, 1998; Hamrick, 2003).

Today's ornamental plant industry offers a large number of plant species and

cultivars for sale including: traditional bedding plants, trees and shrubs, vegetables,

ground covers, and foliage crops. However, from a research and development point of

view, perennials are the most popular crop in the North American market. This broad

classification of herbaceous plants has produced an abundant amount of research and

production information over the past 15 years (Nau, 1999). In 2004 production totals of

perennials were reported at $687 million, up 8% from the previous year (NASS, 2004),

and are the fastest-growing segment of floriculture crops.

As herbaceous perennial production becomes more efficient, plants are able to be

mass produced with less capital input. Thus, perennials are becoming available at the

low priced, "big box" stores, which merchandise nearly 75% of horticultural products

(Wolnik, 1994). Faced with a highly competitive market and narrow profit margins, the

U.S. Bedding Plant Industry has shifted from the traditional packaging of multi-celled,

inexpensive units with many plants per container, to larger pots that fetch a higher price









per individual plant, some of which include: 4.5-inch round-pots, trade-gallon nursery

containers, and various specialty container gardens including color bowls, window boxes,

and patio planters (Ball, 1998; Hamrick, 2003).

This shift has brought an increased interest in using temperate climate herbaceous

perennials for winter landscapes in the South; unfortunately the excessively high

temperatures of summer can negatively impact those plants that perform well under

USDA Hardiness Zones 4 through 7. Hence, tropical perennials are growing in

popularity. These versatile plants can be used as hardy perennials in southern climates

and as summer annuals in the North.

Annuals have often been described as useful enhancements to landscape designs

through foliar textures, flower colors, and planting patterns that can be changed each year

(Arent and Voigt, 1994). Perennial plant viewpoints differ amongst various regions of

the country, and what may be considered a perennial in one location may likely be

considered an annual in another (Nau, 1999). Regardless of these differences in opinion

or performance of perennials, there has been an emerging trend for gardeners to purchase

and trial new and exciting plants.

Because the popularity of gardening with tropical plants has grown in the U.S.,

there has been a considerable increase in production and sales of tropical annuals and

perennials over the past several years (Bowden, 2004). However, little cultural

information is known on how to reproduce these plants effectively, so growers tend to

use some degree of trial and error (G. Griffith, Hatchett Creek Farms, personal

communication). This literature review will focus on how tropical plants are asexually









propagated by stem cuttings, with an emphasis on select tropical species, and how the

stock plants are managed.

Asexual Propagation of Cuttings

Vegetative or asexual propagation has become an established system for

reproducing plants which exhibit desirable characteristics. The common methods for

asexual propagation include using roots, stems, or leaves of stock plants for grafting,

tissue culture, division, or cutting propagation (Davidson et al., 2000). In general,

propagation by stem cuttings has numerous advantages. Many plants can be grown in

high density trays from a limited amount of stock plants. As compared to other asexual

means of reproduction, it is typically less expensive, quicker, relatively simple, and does

not require mastery of difficult techniques such as grafting, budding, or micro-

propagation (Hartmann et al., 2002). Stem cutting production involves removing a shoot

tip from a mother plant and planting it in a growing substrate to root. This method allows

for retention of foliar, flowering, and growth habit characteristics that may not be carried

over via seed production.

Successful propagation of landscape plants by rooting vegetative stem cuttings

depends on several factors, including: physiological state of stem cuttings, the

propagation environment, fertility management, and growth regulator treatments, whether

applied to the stock plant prior to harvesting or exogenously applied rooting hormones to

stem cuttings (Atzmon et al., 1997; Hartmann et al., 2002)

Physiological Age of Cuttings

The physiological age of plant material harvested for propagation has been

documented as an important factor in some tropical species. Terminal stem or tip

cuttings include: the stem apex or shoot tip, developing immature leaves, and one or more









mature leaves. These cuttings generally produce a more efficient crop, by effectively

rooting in the quickest time (Dole and Wilkins, 1999). Older stem tissue is frequently

slower to root than young tissue, but generally cuttings with thick stems and short

internodes will produce the best plants. There is little information published on how to

root tropical cuttings or at which physiological age the tissue should be. However, some

species such as bougainvillea (Bougainvillea glabra Choisy) root better from semi-

hardwood to hardwood cuttings depending on temperature (Schoellhorn and Alvarez,

2002), location (Auld, 1987), and time of year (Chakraverty, 1970).

Growing Environment

Propagation success also depends on the level to which a suitable growing

environment is offered. The propagation substrate temperature, ambient air temperature,

and relative humidity levels should be carefully monitored during propagation, whether

the plant material is herbaceous, softwood, semi-hardwood, or hardwood tissue. Tropical

plants such as bougainvillea benefit from bottom soil temperatures of 30 C (86 F) (Singh

et al., 1976). If bottom heat is provided, soil thermometers or remote recording sensors

should be installed within the root zone area and checked frequently. Excessively high

temperatures in substrate, even for a short time, are likely to result in damage to basal

tissue (Hartmann et al., 2002).

In addition to temperature, it is important to maintain high humidity levels while

rooting vegetative stem cuttings in order to reduce water loss due to transpiration.

Intermittent mist should be used to maintain high humidity levels, but unfortunately

prolonged periods of mist can leach mineral nutrients (Hartmann et al., 2002). In most

cases, cuttings benefit from either a top dressing of slow release fertilizer or a low level

application of liquid fertilizer after roots begin to emerge from the cutting base. Because









some tropical plants often require extensive rooting time in propagation, the risk of air

and waterborne pathogens also increases (Howard, 1994).

Mineral Nutrition

Mineral nutrition is one of many factors influencing formation of adventitious roots

in cuttings. Rooting stages can be generalized into two categories: 1) root initiation; and

2) root growth and development (Blazich, 1988b). Therefore, when considering the

influence various mineral nutrients have on adventitious root formation, one should

consider the function of these elements during each stage of development.

Root initiation involves dedifferentiation of specific cells, leading to the formation

of root meristems and is dependent on the presence of auxin, whether endogenous or

artificially applied. Most studies have not given a clear understanding of the importance

of specific nutrients in the initiation of adventitious roots; however one could argue that

any nutrient essential for root initiation is also necessary for plant growth and

development. For example, nitrogen (N) content is crucial in cuttings as it is important

for nucleic acid and protein synthesis in plant tissue (Hartmann et al., 2002). Nitrogen

concentration also affects cutting yield of stock plants and rooting performance of

cuttings. When considering the role N has in metabolic processes, one could strongly

argue its importance in root initiation (Blazich, 1988b). While excessive N has been

shown to negatively affect cutting propagation (McAvoy, 1995), heavy fertilization in

some tropical plants such as bougainvillea, has been observed to inhibit flowering as it

promotes vegetative shoot growth (Schoellhorn and Alvarez, 2002).

While the role of specific elements in root initiation remains unclear, the nutrition

of stock plants seems to have a more noteworthy effect. Although this influence on stock

plants of tropical perennials is not well documented, nutrient level concentrations









recommended for other floriculture crops have been studied. Dreuge et al. (2000)

reported a positive correlation with the number and length of roots of propagules when

stock plants of chrysanthemum (Dendranthema x grandiflorum Kitam.) were treated with

increased N concentration rates from 60 to 400 mgL-1. In another experiment, Roeber

and Reuther (1982) showed a negative correlation in cutting production with increasing

rates of N fertilization, when concentrations of 56, 112, or 168 mgL-1 were applied to

stock plants of chrysanthemum grown in nutrient solution. In a study where stock plants

of geranium (Pelargonium x hortorum L. H. Bailey) were fertilized at N concentrations

of 50, 100, 200, and 400 mgL-1, respectively, higher concentrations of N produced

similar results in number of vegetative cuttings generated, but with N at 50 mgL-1, lower

numbers of vegetative cuttings developed (Ganmore-Neuman, 1990; Ganmore-Neuman,

1992).

Although optimum nutrient standards for tropical stock plants remain undefined, as

with stock plants from other floriculture crops, it is essential that the propagules come

from a nutritionally healthy source (Hartmann et al., 2002). Nutrition studies on stock

plants of tropical perennials should be conducted to determine optimum concentrations of

N fertilization necessary to produce the greatest number of high quality cuttings with

subsequently greater propagation success and cuttings establishment.

Growth Regulator Treatments

Some naturally occurring plant hormones such as ethylene and cytokinin can be

used as growth regulator treatments to promote lateral branching. The effects plant

growth regulators (PGRs) ethephon [(2-chloroethyl) phosphonic acid] (Florel) (Monterey

Lawn and Garden Products, Inc., Fresno, Calif.) and benzyladenine (BA) (cytokinin)

have on stock plant cutting quantity has been investigated and has been proven to be









beneficial for promoting axillary shoot development in some floricultural crops such as:

chrysanthemum; geranium; poinsettia (Euphorbiapulcharrima Willd. ex Klotsch); and

vinca vine (Vinca major L.) (Konjoian, 1994; Witaszek and Mynett, 1989). In foliar

applications of ethephon to stock plants of Scaevola (Scaevola aemula R. Br.) cutting

length was shown to be longer with ethephon foliar sprays of 250 to 1000 mgL-1 than the

untreated control (Gibson and Whipker, 2004). There is some evidence that cytokinin

applications assist in adventitious root formation. Improved shoot development and

rooting is achieved by its application to leaves and buds of the stock plant, rather than to

the basal tip of stem cuttings (Davis et al., 1988). However in most cases cytokinin

application to plants will inhibit adventitious root formation as it increases lateral

branching (Mynett, 1985).

Rooting Hormone Treatments

Countless studies have reported on the stimulatory influence of auxin on the

propagation of cuttings of difficult-to-root species. Treating cuttings with auxin increases

rooting percentage, hastens root formation, and increases uniformity of rooting (Davis et

al., 1988). Indole acetic acid (IAA) is the naturally occurring auxin found in plants and

has been documented in nearly every aspect of plant growth and development. Some of

the processes regulated by IAA, as they pertain to tropical plant propagation include:

induction of cell division, stem elongation, apical dominance, induction of rooting, and

vascular tissue differentiation (Srivastava, 2002). Synthetic forms of auxin are

commercially available in the form of indolebutyric acid (IBA) and napthaleneacetic acid

(NAA). Preference for these sythentic compounds compared to IAA, is illustrated by the

large number of rooting products containing IBA, NAA, or in combination (Blazich,

1988a).









Auxin containing rooting hormones can be applied to the base of cuttings as talc-

based powder or dipped in a liquid solution containing 50% ethanol or isopropyl alcohol

and 50% water. Cuttings are dipped for a period of time from a few seconds to 12 hours

(Dole and Wilkins, 1999). Liquid treatments usually provide the most consistent results

because application is more uniform than with powder. However, powder dips are

sometimes preferred since it decreases the chance for water-borne disease transmittance.

There are also potassium salt (K+) formulations that enable IBA (KIBA) and NAA

(KNAA) to be dissolved in water (Hartmann et al., 2002).

Sometimes mixing auxin with other carriers such as polyethylene glycol

(antifreeze), propylene glycol, or windshield washer fluid may facilitate auxin application

on difficult-to-root species which require high levels of hormone to induce rooting, but

with sensitivity to alcohol. However these methods often involve heating the solutions to

72 C (161.6 F), which facilitates dissolving the auxin into polyethylene glycol causing

the materials to not disassociate after cooling (Hartmann et al., 2002). Often nurseries

use KIBA with vegetative cuttings during active growth stages and IBA with ethanol

during dormant periods.

Research has suggested some tropical species such as oleander (Nerium oleander

L.) and bougainvillea benefit from applications of 3,000 mgL-1 IBA and 4,000 to 16,000

mgL-1 IBA respectively (Hartmann et al., 2002). In other difficult-to-root tropical

species an evaluation of commercially available rooting hormones should be conducted

to determine the optimum concentration to achieve the highest rooting percentage.

Conclusion

Shortages in cutting availability and poor cutting quality exists among shipments to

growers, subsequently low quality rooted liners supplied by domestic propagation firms









has become a concern for growers wishing to produce tropical plants for sale. Examples

include poorly rooted or nutrient deficient plants with weak stems and poor lateral

branching. Propagators of tropical plants need to adopt nutrient management and PGR

treatment programs which produce the maximum quantity of high quality cuttings so that

growers who receive rooted liners can profit from the specialized cultivation of the stock

plants. Relating shoot and root performance of cuttings from stock plants fertilized at

different rates of liquid fertilizers and treated with lateral branching agents is a means of

achieving this goal. Additionally identification of optimum rooting hormone

concentrations is essential to the efficient production of high quality rooted liners.

Selected Tropical Species

To better understand the significance of selected tropical perennials in this review

and purpose for which they are being examined, detailed descriptions of each are as

follows:

Allamanda schottii

Allamanda or Golden Trumpet (Allamanda schottii L.) is a member of the family

Apocynaceae or Milkweed Family which contains other important ornamental plants

such as flowering vinca (Catharanthus roseus L.), milkweed (Asclepias incartata L.),

wax plant (Hoya carnosa L. f.), oleander, frangipani (Plumeria alba L.), and vinca vine

(Judd et al., 2002). Allamanda is characterized by its showy, trumpet-shaped, yellow to

purple-red flowers and is native to Tropical America (Griffiths, 1994). Depending on

species, allamanda may be used as a climbing vine or as a shrub. It performs well in

neglected, poor soil and is vigorous enough to withstand coastal conditions (Courtright,

1988; Walker and Hanly, 1996). Certain species of allamanda are reported to propagate

easily by semi-hardwood cuttings in summer and fall with up to 2,000 mgL-1 IBA









applied to the cutting base (Hartmann et al., 2002). There is little information published

on propagation during other seasons, using other combinations of rooting hormones, or

stock plant treatments to improve cutting quantity and quality.

Bougainvillea glabra

Bougainvilleas are members of the family Nyctaginaceae or the Four O'Clock

Family, which includes other ornamentals such as four o'clock (Mirabalis longiflora L.)

and sand verbena (Abronia latifolia Eschsch.) (Judd et al., 2002). Bougainvillea is native

to South America and was originally collected by Commerson, a French Botanist from

Brazil. Commerson named the plant, because of its wandering growth habit, after Louis

Antonyne de Bougainville, a French navigator known for his journey around the world

near the end of the 18th century (Rama Rao, 1976).

Bougainvillea are characterized by colorful bracts which exhibit the best flowering

performance during cool season months and shorter day lengths in the U.S. The leaves

are simple and alternate, but shape and texture differ in varieties. The plants can be

grown as specimen shrubs, hedges, pot crops, or climbing vines. They grow readily in

tropical and sub-tropical areas, and are seen in diverse locations from Florida to

California, India, and Australia (Hackett et al., 1972; Rama Rao, 1976; Auld, 1987).

Bougainvillea plants are generally propagated from stem cuttings or air-layerings,

but the root systems are extremely fine and fragile, with low rooting success, even during

the cultivated seasons (Chakraverty, 1970). It has been suggested that the reason

bougainvillea is not more commercially available is due to its difficulty in propagation

(Czekalski, 1989).

The published literature offers varying information on length of stem cuttings and

position. Some suggested cutting lengths include: 10 to 12 cm (Aldrich and Norcini,









1995; Atzmon et al., 1997) 15 to 17 cm (Auld, 1987); 20 to 25 cm (Chakraverty, 1970;

Yadav et al., 1978) and 80 to 100 cm with top portions removed back to 15 cm of total

length (Mudge et al., 1995). Another, perhaps more effective means determining cutting

length in bougainvillea is by counting number of nodes. This process has less of a

widespread definition of parameters, ranging from 2 nodes (Auld, 1987), 3 to 4 nodes

(Atzmon et al., 1997), or 5 to 9 nodes (Schoellhorn and Alvarez, 2002). Most sources

agree that cuttings should be severed 0.5 cm below the bottom node.

Bougainvillea cuttings should be taken from wood that is mature to the point where

it starts to become stiff. If propagating by softwood cuttings, night temperatures should

be above 12.8 C (55 F), whereas hardwood cuttings root more efficiently when night

temperatures are below 12.8 C (55 F) (Schoellhorn and Alvarez, 2002). Cuttings should

be provided with rooting substrate temperatures of 30 C (86 F) (Singh et al., 1976).

Mandevilla splendens

Mandevilla (Mandevilla splendens Hook.f.), like allamanda, is a member of the

Apocynaceae family. It is an evergreen climbing vine characterized by large pink to

white trumpet-shaped flowers appearing mid to late-summer in the U. S. The twining

woody tissue should be supported via trellis or other structure. Mandevilla is native to

Brazil, but is hardy to USDA Hardiness Zone 10 in protected locations (Courtright, 1988;

Griffiths, 1994). Mandevilla, formerly known as Dipladenia, has been documented as a

popular greenhouse plant for over a century in Great Britain (Dress, 1974). Hartmann et

al. (2002) suggest propagating Mandevilla cultivar Alice DuPont with one node and half

of the leaves removed. The stem cutting should then be dipped in 2,500 mgL-1 IBA and

rooted in summer. Other than 'Alice DuPont', there is little published on how to

effectively grow and reproduce Mandevilla from stem cuttings.









Nerium oleander

Oleander is also a member the family Apocynaceae. It should be noted that nearly

all members of this taxa are poisonous (Judd et al., 2002), therefore care should be taken

when harvesting stem cuttings, handling finished plants, and planting and maintaining

specimens in garden designs. Hands should be washed thoroughly and care should be

taken to avoid sap entering the body.

Oleander is an evergreen shrub native to the Mediterranean regions of Southern

Europe and North Africa, where it is widely used outside in landscaping designs and as

an indoor potted plant (Arias et al., 2001). It has been documented that oleander first

arrived in the U.S. via Spanish settlers in St. Augustine, FL, and it was likely that these

plants arrived as stem cuttings (Popenoe, 1976).

Oleander is a versatile tropical perennial, hardy to USDA Hardiness Zone 9 and

which can be used as a foundation plant, hedge, or as a featured specimen plant. Once

established, oleander produces flowers with virtually no maintenance requirements. It

tolerates poor soil conditions and is reportedly salt tolerant (Walker and Hanly, 1996). It

requires a sunny location for northern gardeners, who could use it as a potted plant to be

taken indoors during winter months (McCulla, 1973). Oleander is commonly available as

an evergreen shrub reaching maturity at 6 m (20 ft), with single, sometimes double,

flowers in pink, white, rose, or red. Dwarf varieties are also available (Courtright, 1988).

Propagation has reportedly been successful from cuttings taken from hardwood or

softwood during the summer, at lengths of 15.24 cm (6 in), with quick dip applications of

3,000 mgL-1 IBA (McCulla, 1973; Hartmann et al., 2002). There is limited literature

currently available on how to manage the stock plants of these species and propagation









during other seasons. However, oleander is reportedly not susceptible to ethylene

branching treatments (Dole and Wilkins, 1999).

Tecoma stans

Texas Star [Tecoma stans (L.) Juss.] or (Bignonia stans L.), is a member of the

Bignoniaceae or Trumpet Creeper family, which includes other known ornamentals as

catalpa (Catalpa bignonioides Walter.), sausage tree (Jacaranda arborea Urban.), and

cape honeysuckle (Tecomaria capensis Spach.) (Judd et al., 2002). Tecoma is a

mounding evergreen shrub native to Central America and is characterized by its funnel-

shaped yellow flowers throughout summer and early fall in the subtropics (Walker and

Hanly, 1996). In Florida, Tecoma is a deciduous shrub or tree hardy to USDA Hardiness

Zone 10, that grows to 7.6 m (25 ft) at maturity, unless pruned, and flowers only for a

short period in autumn (Courtright, 1988). Little information is published on how to

commercially produce Tecoma or how to manage stock plants for improved cutting

quantity and quality.

Objectives

1. Determine which of three N liquid fertilizer concentrations is optimal for producing
the greatest number of bougainvillea cuttings with the highest rooting percentage
and quality This study will also simulate long distance shipping, as the cuttings
will be harvested in one location and sent to another for evaluation.

2. Determine which of three concentrations of three commercially available lateral
branching PGRs applied to stock plants will most efficiently increase cutting
quantity and quality, while not adversely affecting subsequent rooting of cuttings.
PGRs surveyed will compare three concentrations of chemicals containing
cytokinin, ethylene, or gibberellin and cytokinin in combination.

3. Determine the optimum concentration of liquid KIBA, delivered via quick dip, for
improving both quantity and quality of adventitious root formation in select
difficult-to-root tropical perennials. KIBA will also be compared to untreated
control and commercial standard quick-dip Dip N' Grow (Dip N' Grow Inc.,
Clackamas, Ore.)(Indole-3-butyric acid 1%, Naphthalene acetic acid 0.5%)













CHAPTER 2
STOCK PLANT NUTRITION

Introduction

Because the popularity of gardening with tropical annuals and perennials has grown

in the U.S., there has been a considerable increase in production and sales of tropical

plants over the past several years (Bowden, 2004). Shortages and poor quality cuttings

supplied by domestic propagation firms have become concerns for growers wishing to

produce tropical plants for sale. Propagators have expressed a need for standardized

production practices to maximize cutting yield and subsequent adventitious root

formation in propagules; however, little is published on how to effectively manage

tropical stock plants (Hartmann et al., 2002).

A tropical species that has potential to be more widely used by gardeners is

bougainvillea (Bougainvillea glabra Choisy). This multi-use plant grows best in U.S.

landscapes during summer months followed by attractive bracts displayed during natural

short-days. Unfortunately because of its warm temperature production requirements and

difficulty in propagation, commercial availability is limited (Czekalski, 1989).

Bougainvillea is generally propagated by stem cuttings or air-layering, yet the rooting

percentage is low, even during the cultivated seasons (Chakraverty, 1970). The root

systems are also known to be extremely fine and fragile (Auld, 1987). Bougainvillea root

best from semi-hardwood to hardwood cuttings depending on temperature (Schoellhorn,

2002), location (Auld, 1987), and time of year (Chakraverty, 1970). These plants have









been documented to benefit from bottom heat with rooting substrate temperatures of 30 C

(Singh et al., 1976).

Nitrogen (N) concentration is crucial in cuttings as it is important for nucleic acid

and protein synthesis in plant tissue (Hartmann et al., 2002). When considering the role

N has in these metabolic processes, one could strongly argue its importance in root

initiation (Blazich, 1988). Nitrogen concentration also affects cutting yield and rooting

performance of cuttings. While excessive N may negatively affect cutting propagation

(McAvoy, 1995), heavy fertilization in some tropical plants such as bougainvillea, has

been observed to inhibit flowering and promote growth (Schoellhorn, 2002).

Although optimum nutrient levels of tropical stock plants remain undefined, it is

crucial that propagules come from a nutritionally healthy source (Hartmann et al., 2002).

Limited research has been conducted in the management of stock plants of bougainvillea

for purposes of maximizing quantity and quality of cuttings suitable for propagation;

therefore the experimental objective was to evaluate the impact of three nitrogen

concentrations on stock plant yield and cutting performance of Bougainvillea 'Purple

Small Leaf and 'Raspberry Ice'.

Materials and Methods

Experiment 1

Rooted stem cuttings of Bougainvillea glabra 'Raspberry Ice' and 'Purple Small

Leaf were transplanted, three per pot, into 2.8-L (0.74-gal) [15.2-cm (6-inch diameter)]

round plastic containers on 18 April 2005. The root substrate was Fafard 2 (Fafard,

Anderson, S.C.), which contained 6.5 sphagnum peat: 2 perlite : 1.5 vermiculite (v/v). A

continual liquid fertilization program was initiated on 23 May, at which time a soft pinch

was provided to develop a canopy for harvesting cuttings. Fertilization treatments were









applied with N concentration levels at 100, 200, or 300 mgL-1. Phosphorous (P),

potassium (K), calcium (Ca), and magnesium (Mg) were held constant at 20, 200, 100,

and 50 mgL-1, respectively. Micronutrients were provided using soluble trace element

mix in solution (S.T.E.M.) (The Scotts Company, Marysville, Ohio) at a rate of 6.78

mgL-1. Supplemental iron was applied using ferric ethylenediaminetertraacetic acid

(FeEDTA) (Sequestrine) at a rate of 1.76 mgL-1. Ammoniacal-nitrogen was 6% to 35%

of the total nitrogen in all treatments. Plants were fertigated through a drip irrigation

system. The experiment was a randomized complete block design with 12 single-plant

replications of each of three treatments. Stock plants were grown in a greenhouse under

natural photoperiod and irradiance with day/night minimum setpoint temperatures of

21/18 C.

Leachate samples were collected initially 4 weeks after potting and biweekly

thereafter until 28 weeks after potting via a modified Virginia Tech Extraction Method

(VTEM) (Wright, 1986) to determine pH, electrical conductivity (EC), and nitrate

nitrogen (NO3-N). Distilled water (85 ml) was applied to the substrate surface to displace

50 ml of leachate. Leachate solutions were analyzed for pH (pHep pH meter; Hanna

Instruments, Woonsocket, R.I.), EC (DiST WP4 EC meter; Hanna Instruments), and

NO3-N (Horiba Compact Ion Meter; C-141 Spectrum Technologies, Inc., Plainfield, Ill.).

Leachate was collected from 3 random experimental blocks at each sampling date and

values were recorded as replications 1, 2, and 3, respectively. Photosynthetic photon flux

(PPF) was recorded each day at or near solar noon using a Quantum Meter (Spectrum

Technologies, East-Plainfield, Ill.). Average monthly PPF readings were 453, 586, 426,

596, 647, 355, and 380 imol.m-2S-1, for June through Dec., respectively.









Stock plants were maintained in Milton, Fla. and cuttings were harvested every

three weeks starting 7 weeks after potting, on 7 June (HI), 28 June (H2), 19 July (H3), 9

Aug. (H4), 30 Aug. (H5), and 20 Sept. (H6), respectively. Harvesting protocols consisted

of removing all sub-terminal stem cuttings from stock plants by excising the shoot tip 0.5

cm above the first fully mature leaf node, and then harvesting the cutting 1 cm below the

fourth successive node (Cerveny et al., 2005).

The quantity of cuttings was recorded at each harvest. At 7, 13, and 19 weeks after

potting shoot length and diameter was measured at the base of 3 randomly selected

cuttings per plant. Cutting leaf area was also measured with a LI-COR LI-3100 portable

area meter (LI-COR, Lincoln, Nebr.). For tissue analysis, leaves from all cuttings were

removed and stock plant blocks 1 to 4, 5 to 8, and 9 to 12 were pooled into treatment

replications 1, 2, and 3, respectively. Tissue samples were rinsed in deionized water,

then washed in 0.2 N HCl for 30 s, and dried at 70 C for 72 h. Dried tissue was ground

in a stainless steel Wiley mill to pass a 1-mm screen (20-mesh). Tissue was then

analyzed for macro and micronutrients at Quality Analytical Labs (QAL, Panama City,

Fla.).

Cuttings harvested 10, 16, and 22 WAP were packaged in moist towels and inserted

into perforated plastic bags. The bags were placed into boxes, along with a temperature

and relative humidity sensor (Hobo Pro, model number H08-032-08) (Onset Computer,

Bourne, Mass.), then stored over-night in a cooler at 10 C. Boxes were sealed and

cuttings were then shipped 24h, to simulate industry handling. Upon arrival, boxed

cuttings were left intact until the following morning and were then unpacked and planted,

totaling approximately 72 h between cutting excision and planting (Table 2-1).









Cuttings harvested from stock plant replications 1 to 12 of each treatment were

pooled and assigned randomly to 6 propagation replicates with 6 sub-samples of each

treatment. Basal portions of cuttings (1 cm) were inserted into indole-3-butyric acid,

potassium salt (KIBA) at 3,000 mgL-1 for 3 s then planted into 3.3 x 8.8 x 6 cm 6-cell

containers in a substrate consisting of 4 perlite: 1 vermiculite (v/v) and placed under

intermittent mist. Cuttings were maintained under natural photoperiod with day/night

set-point temperatures of 20/18 C, and misted daily for 5 s every 20 min (07:00 19:30).

Minimum rooting substrate temperatures were provided at 24 C and bottom heat was

supplemented via rooting mats as needed. The experimental design was a completely

randomized design with 2 cultivars, 3 N treatments, and 6 replications per treatment with

6 sub-samples (pooled) per replicate.

Adventitious roots > 1.0 mm in length were evaluated 4 and 6 weeks after planting

(WAP). At the first evaluation of each harvest percent survival (based on presence of at

least minimal callus formation), rooting quality, number of primary roots, number of

lateral roots, cutting total dry weight, and cutting root : shoot ratio (root weight divided

by shoot weight) were recorded. At the second evaluation, roots were not counted; all

other parameters were recorded in the same manner as the first. Rooting quality was

based on a 1 to 5 scale: 1= minimal callus formation with no roots; 2= minimal rooting;

3= moderate, uneven rooting; 4= moderate, uniform rooting; 5= well-developed rooting.

Cuttings were prepared for dry weight measurement by excision of the rooted

portion of stem 1 cm above cutting base. Then each portion was placed into individual

envelopes and dried for 3 d at 70 C. All data were subjected to analysis of variance using









generalized linear model procedure in SAS (SAS institute, Cary, N.C.) and means

separated using LSD with P < 0.05.

Experiment 2

The second experiment was replicated in the same manner as the first experiment,

with the following exceptions: Stock plants were transplanted 18 July, and given a soft

pinch along with initiation of fertilization treatments on 8 Aug. 2005. Cuttings were

harvested every three weeks on 23 Aug. (HI), 13 Sept. (H2), 4 Oct. (H3), 25 Oct. (H4),

15 Nov. (H5), and 6 Dec. (H6). Cuttings were handled in the same manner as the first

experiment, except they were shipped to Gainesville, Fla. using Federal Express 24 h

service (Table 2-1).

Results

Experiment 1

Stock Plant Parameters

Cutting quantity

The harvest date x cultivar and harvest date x N concentration interactions were

significant for cutting quantity (Table 2-2). For 'Raspberry Ice', cutting quantity at HI

(10.2) was similar to all other harvest dates. Harvest 4 and H6 were similar to each other

with 8.9 and 8.2 cuttings produced per plant, respectively. Both H4 and H6 were at least

28% less than H2, H3, or H5, all of which produced 12.0 cuttings per plant (LSD=2.4,

n=36). 'Purple Small Leaf produced a similar number of cuttings at HI and H6 (17.0 and

17.4, respectively), while both were at least 30% less than all other harvests. Harvest 5

produced more cuttings (31.3) than H2 (24.8) or H3 (25.1) (LSD=5.2, n=36). Harvest 4

produced a similar number of cuttings to H2, H3, and H5. For all harvests, 'Purple Small

Leaf produced more cuttings per plant, than 'Raspberry Ice' (data not shown). When









considering both cultivars, differences in cutting quantity were not significant amongst

treatments for HI, H2, or H4, with 13.5, 18.4, and 18.9 cuttings produced per plant,

respectively (Table 2-2). For H3, 126% more cuttings were harvested from stock plants

fertilized with N at 300 mg-L-1 than from N at 200 or 100 mg-L-1. For H5, stock plants

fertilized with N at 200 mg-L-1 produced a similar number of cuttings to those fertilized

with N at 300 mg-L-1, while both concentrations produced at least 153% more cuttings

than N at 100 mg-L-1. For H6, N at 300 mg-L-1 produced more cuttings than at 100 mg-L-

1, while both treatments were similar to N at 200 mg-L-1. For stock plants fertilized with

N at 100 or 300 mg-L-1, differences in cutting quantity were not significant amongst

harvests, with 14.4 and 19.7 cuttings produced per plant, respectively. For stock plants

fertilized with N at 200 mg-L-1, the number of cuttings produced at H5 was similar to H4

and H2, yet greater than HI, H3, and H6.

Cutting length, stem diameter, and leaf area

There was a harvest date x N concentration interaction for cutting length; the main

effect for cultivar will be presented separately (Table 2-3). Stock plants of 'Raspberry

Ice' and 'Purple Small Leaf produced similar length cuttings (7.0 cm). For HI and H5,

differences in cutting length were not significant amongst treatments with lengths of 7.0

and 7.5 cm, respectively (Table 2-4). At H3, stock plants treated with N at 100 or 200

mg-L-1 produced cuttings that were at least 112% longer than those treated with N at 300

mg-L-1. Over time cuttings harvested from stock plants treated with N at 100 mg-L-1 had

longer cuttings at H5 than at H3, while both harvest dates were similar to HI. Stock

plants of bougainvillea produced cuttings with a mean stem diameter of 0.24 cm and

differences were not significant amongst treatments.









There was a cultivar x N concentration interaction for leaf area (Table 2-3). The

harvest date main effects will be presented separately. Cuttings from H5 (40.5 cm2) had

a 114% larger leaf area than H1 (35.3 cm2), and was 116% larger than H3 (30.4 cm2)

(LSD=2.1, n=72). Stock plants of 'Raspberry Ice' fertilized with N at 100 mg-L-1

produced cuttings with similar leaf area to 200 mg-L-1 (45.1 cm2), while both

concentrations were greater than N at 300 mg-L-1 (38.9 cm2) (LSD=4.3, n=36).

Differences in leaf area were not significant amongst treatments for stock plants of

'Purple Small Leaf.

Rooting Parameters

Percent survival, visual rooting quality, and quantity of primary and lateral roots

There was a cultivar x N concentration and a harvest date x N concentration

interaction for percent survival of planted cuttings (Table 2-5). Differences in percent

survival were not significant for 'Raspberry Ice' and 'Purple Small Leaf (84% and 58%,

respectively). 'Raspberry Ice' had a higher percent survival than 'Purple Small Leaf when

stock plants were fertilized with N at 100 to 300 mg-L-1 (data not shown). When

considering both cultivars, cuttings planted from H2 had higher percent survival from

stock plants provided with N at 300 mg-L-1 than from concentrations of 100 to 200 mg-L-

1 (Table 2-6). Differences in percent survival were not significant amongst treatments for

H4 or H6. Nitrogen fertilization at 100 mg-L-1 produced cuttings with a higher percent

survival at H2 or H4 than at H6. Differences in percent survival were not significant

amongst harvest dates, with 70% survival when stock plants were fertigated with N at

200 mg-L-1. Similar to N at 100 mg-L-1, stock plants fertigated with N at 300 mg-L-1 had

a higher percent survival from H2 or H4 than from H6.









There was a harvest date x cultivar x N concentration interaction for rooting quality

(Table 2-5). The main effects for evaluation week will be presented separately. Cuttings

evaluated 6 WAP had a root quality rating of 2.2, which was higher than those evaluated

4 WAP (2.0) (LSD=0.2, n=108). For H2, differences in rooting quality were not

significant amongst treatments for cuttings of 'Raspberry Ice' (Table 2-7). 'Purple Small

Leaf, however, had lower rooting quality when provided N at 100 or 200 mg-L-1 as

compared to 300 mg-L-1. For H4, differences in rooting quality for both 'Raspberry Ice'

and 'Purple Small Leaf were not significant amongst treatments. For H6, 'Raspberry Ice'

cuttings planted from stock plants treated with N at 200 mg-L-1 had higher rooting quality

than from those treated with N at 100 or 300 mg-L-1. Differences in rooting quality for

'Purple Small Leaf cuttings planted from H6 were not significant amongst treatments.

When considering the interaction over time, 'Raspberry Ice' exhibited a reduction in

rooting quality with each harvest of stock plants treated with N at 100 to 300 mg-L-1.

Differences in rooting quality for 'Purple Small Leaf were not significant over time for

most treatments, except with N at 300 mg-L-1, which had higher rooting quality from H2

than from H4 and H6. 'Raspberry Ice' had higher rooting quality than 'Purple Small Leaf

for all treatments and harvests, except for cuttings planted from stock plants treated with

N at 300 mg-L-1 and harvested at H2, or any concentration of N at H6, in which the two

rooted similarly.

There was a harvest date x cultivar and a cultivar x N concentration interaction for

number of primary roots (Table 2-8). For cuttings planted from H2, H4, and H6,

'Raspberry Ice' had a greater number of primary roots than 'Purple Small Leaf (data not

shown). Additionally 'Raspberry Ice' cuttings planted from H2 produced 14.9 primary









roots per cutting, which was at least 470% more than from H4 (3.2) or H6 (1.2)

(LSD=2.8, n=18). 'Purple Small Leaf also had a similar number of primary roots at H4

and H6 (0.3), both of which were less than H2 (2.7) (LSD=1.4, n=18).

When comparing cultivars, stock plants of 'Raspberry Ice' fertilized with N at 100

mg-L-1 had a greater number of primary roots (5.5), than 'Purple Small Leaf (1.1)

(LSD=3.0, n=18). Similarly, when fertilized with N at 200 mg-L-1, stock plants of

'Raspberry Ice' produced cuttings with greater number of primary roots than 'Purple

Small Leaf (data not shown). When fertilized with N at 300 mg-L-1 differences between

cultivars were not significant, with 3.8 roots produced per cutting.

There was a harvest date x cultivar x N concentration interaction for number of

lateral roots (Table 2-8). The number of lateral roots on cuttings planted from H4 and H6

was not significantly different amongst treatments, with 6.2 and 2.0 lateral roots for

'Raspberry Ice', and with 0.1 and 0.7 lateral roots for 'Purple Small Leaf, respectively.

Cuttings of 'Raspberry Ice' planted from H2 had a greater number of lateral roots when

fertilized with N at 200 mg-L-1 (37.2) than at 100 mg-L-1 (19.7), however both were

similar to N at 300 mg-L-1 (29.7) (LSD=12.2, n=6). 'Purple Small Leaf cuttings planted

from H2 produced a greater number of lateral roots from stock plants treated with N at

300 mg-L-1 (10.4) than either 100 or 200 mg-L-1 (2.5 and 0.2, respectively) (LSD=4.1,

n=6). Both cultivars produced a similar number of lateral roots (1.7) with cuttings

planted at H4 from stock plants treated with N at 300 mg-L-1 (data not shown). Similarly,

for cuttings planted from H6, differences between cultivars were not significant for all N

concentrations. 'Raspberry Ice' cuttings had a greater number of lateral roots than 'Purple









Small Leaf when planted from stock plants treated with any N concentration from H2

and N at 100 to 200 mg-L-1 from H4.

Cutting dry weight and root : shoot ratio

There was a harvest date x cultivar x N concentration and a harvest date x

evaluation week x cultivar interaction for total cutting dry weight (Table 2-5). 'Raspberry

Ice', cuttings planted from H2 had at least a 17% lower total dry weight when propagated

from stock plants treated with N at 100 mg-L-1 than at 200 or 300 mg-L-1 (Table 2-9). For

cuttings planted from either H4 or H6, differences in total dry weight were not significant

amongst treatments with 165 and 223 mg per cutting, respectively. 'Purple Small Leaf

cuttings propagated at H2 from stock plants treated with N at 300 mg-L-1 had at least

133% greater dry weight than 100 or 200 mg-L-1. Differences in total dry weight were

not significant amongst N concentrations for H4, with 165 mg total dry weight per

cutting. From H6, cuttings propagated from stock plants treated with N at 100 to 200

mg-L-1 had greater dry weight than at 300 mg-L-1. For each N concentration provided and

with all harvest dates, 'Raspberry Ice' cuttings had greater dry weight than 'Purple Small

Leaf when planted from all harvest dates and for each N concentration applied to stock

plants.

'Raspberry Ice' cuttings evaluated from H2 had a greater dry weight 6 WAP than 4

WAP (257 vs 213 mg, respectively) (LSD=29, n=18). Similarly, H4 cuttings evaluated 6

WAP had higher dry weight than 4 WAP (data not shown), however at H6 those cuttings

evaluated 6 WAP had an 11% lower dry weight than 4 WAP (235 vs 210 mg,

respectively) (LSD=22, n=18). 'Purple Small Leaf cuttings also had higher dry weight 6

WAP as compared to 4 WAP for those planted from H2 (160 vs 132 g) (LSD=27, n=18).









Yet from H4 and H6, differences in total dry weight were not significant between

evaluation weeks, with 101 and 132 mg produced per cutting, respectively.

There were harvest date x evaluation week x cultivar, harvest date x evaluation

week x concentration, and cultivar x N concentration interactions for the cutting root:

shoot ratio (Table 2-5). 'Raspberry Ice' cuttings had a higher root : shoot ratio than

'Purple Small Leaf for each concentration of N applied, however differences in root:

shoot ratio were not significant amongst treatments applied to 'Raspberry Ice' with a

mean ratio of 0.26 per cutting. 'Purple Small Leaf had a higher root: shoot ratio with N

at 300 mg-L-1 than at 200 mg-L-1 (0.18 vs 0.14), while both were similar to 100 mg-L-1

(0.15) (LSD=0.04, n=36). While 'Raspberry Ice' had a higher root: shoot ratio than

'Purple Small Leaf for H2 and H4 at both 4 and 6 WAP (data not shown), cuttings

planted from H6 had a similar root : shoot ratio between cultivars of 0.15 and 0.14 at 4

and 6 WAP, respectively.

When considering both cultivars differences in root : shoot ratio were not

significant amongst treatments or evaluation weeks for H2, H4, and H6, except cuttings

evaluated 6 WAP from H6. Cuttings planted from H6 had a greater root : shoot ratio 6

WAP from stock plants treated with N at 200 mg-L-1 (0.17) than from N at 300 mg-L-1

(0.11), while both were similar to 100 mg-L-1 (0.14) (LSD=0.03, n=12). When evaluated

4 WAP, cuttings planted from N concentrations of 100 or 200 mg-L-1 showed similar root

: shoot ratios amongst harvest dates. When N was applied at 300 mg-L-1, cuttings

evaluated 4 WAP had a higher root: shoot ratio at H2 than H4 or H6 (data not shown).

For cuttings evaluated 6 WAP, those propagated from stock plants treated with N at 100

mg-L-1 had a higher root: shoot ratio from H2 (0.27) or H4 (0.23) than H6 (0.14)









(LSD=0.09, n=12). Cuttings planted from stock plants fertigated at 300 mg-L-1 produced

similar root: shoot ratios with H2 or H4 and greater than H6 (data not shown), however

cuttings planted from stock plants treated with N at 200 mg-L-1 were similar amongst

harvest dates with a root : shoot ratio of 0.24.

Stock plant leachate

There was a sampling date x N concentration interaction of leachate pH; main

effects for cultivar will be presented separately. The root substrate of stock plants of

'Raspberry Ice' had a mean leachate pH of 5.3, which was greater than that of 'Purple

Small Leaf (5.1). Generally pH values decreased with increasing concentrations of N

applied to stock plants (Figure 2-1). Variations occurred over time and increased

concentrations of N applied to stock plants tended to reduce pH values below acceptable

values of 5.5 to 6.0 (Dole and Wilkins, 2005). Although pH values were in the

acceptable range with N at 100 mg-L-1, rooting was not improved.

The sampling date x N concentration interaction was not significant for electrical

conductivity (EC) therefore the main effects will be presented. Overall, EC values

remained within the recommended range of 2.0 to 3.5 mS/cm for producing bougainvillea

plants established by Whipker et al. (2003) (Figure 2-2). A decling then increase in

soluble salts was observed during H2 to H4, suggesting nutrients were utilized for growth

of new shoots or leached. As the concentration of N delivered to stock plants increased,

so did leachate EC. Stock plants fertigated with N at 300 mg-L-1 had the highest EC

values of 3.84 mS/cm, while N at 200 mg-L-1 (2.92 mS/cm) was greater than 100 mg-L-1

(1.87 mS/cm) (LSD=0.56, n=48). When comparing cultivars, 'Raspberry Ice' (3.24

mS/cm) had higher EC values than 'Purple Small Leaf (2.52 mS/cm) (LSD=0.46, n=72).









There was a sampling date x cultivar interaction for nitrate nitrogen (NO3-N); main

effects for N concentration will be presented separately. Generally NO3-N values were

shown to increase with removal of plant tissue during cutting harvests (Figure 2-3). This

trend was more evident in stock plants of 'Purple Small Leaf than 'Raspberry Ice',

although overall leachate NO3-N values tended to be higher for 'Raspberry Ice'. As N

concentration increased, NO3-N values also increased, and were highest from stock plants

fertigated with N at 300 mg-L-1 (391.9 mg-L-) while N at 200 mg-L-1 (278.5 mg-L-) was

greater than at 100 mg-L-1 (179.8 mg-L-) (LSD=83.9, n=48).

Foliar analysis

There was a harvest date x cultivar interaction for foliar N, phosphorous (P),

potassium (K), calcium (Ca), and Magnesium (Mg); a harvest date x N concentration

interaction for foliar N, P, Ca, and Mg; and a cultivar x N concentration interaction for

foliar P, K, and Mg (Table 2-10). 'Raspberry Ice' was generally similar to 'Purple Small

Leaf when comparing foliar N or Mg concentrations, although when significant

'Raspberry Ice' tended to be greater than 'Purple Small Leaf (Table 2-11). For K and P

tissue concentrations, 'Purple Small Leaf tended to be greater than 'Raspberry Ice' at each

harvest, while P tissue concentrations tended to be greater for 'Raspberry Ice' than 'Purple

Small Leaf.

Generally foliar N concentration was similar with each concentration of N provided

to stock plants, however when cuttings were analyzed from H5, foliar N concentration

increased as N concentration increased (Table 2-12). For foliar P, all N fertigation

concentrations generally had a similar percentage, except those cuttings analyzed from

H3, where P levels decreased as N concentration increased. Both foliar Ca and Mg

concentrations decreased with increasing concentrations of N applied. This decrease may









be due to the low pH substrate inhibiting uptake of these nutrients (Whipker et al., 2003).

Magnesium levels remained within acceptable ranges of 0.2% to 0.8% as recommended

by Dole and Wilkins (2005); however Ca levels fell below the 1.0% to 2.0%

recommendation, when the N concentration increased to 300 mg-L-1 (Table 2-12).

When tissue data was compared for each cultivar, both had similar results for P

levels (Table 2-13). As the N concentration delivered to stock plants increased, foliar K

concentration decreased; this trend occurred at lower N concentrations for 'Purple Small

Leaf than for 'Raspberry Ice'. For foliar Mg concentration, 'Raspberry Ice' levels

decreased with increased concentrations of N applied, while 'Purple Small Leaf had

higher Mg levels with N at 200 mg-L-1 than at 100 or 300 mg-L-1.

Experiment 2

Stock plant parameters

Cutting quantity

There was a harvest date x cultivar and a cultivar x N concentration interaction for

cutting quantity (Table 2-14). Harvest 3 produced the most cuttings of 'Raspberry Ice' at

9.2 per plant, while HI (4.9), H2 (5.9), H4 (4.6), and H5 (5.4) all produced similar

quantities, which were at least 157% more than H6 (2.9) (LSD=1.4, n=36). For 'Purple

Small Leaf, cutting quantity was similar at H4 (21.7) and H3 (20.7), both of which were

greater than all other harvest dates. Harvest 2 (16.3) produced more cuttings than H5

(12.9), yet both were similar to H6 (15.4). Harvest 1 produced 7.6 cuttings per plant,

which was at least 42% less than any other harvest (LSD=3.2, n=36). When comparing

cultivars, 'Purple Small Leaf produced cuttings in greater quantity than 'Raspberry Ice'

during all harvests (data not shown).









'Purple Small Leaf stock plants produced more cuttings that 'Raspberry Ice' for all

N concentrations applied (Table 2-14). 'Raspberry Ice' stock plants produced more

cuttings with N at 300 mg-L-1 than at 100 or 200 mg-L-1. For 'Purple Small Leaf, cutting

quantity was greater for stock plants treated with N at 200 mg-L-1 than at 100 mg-L-1

while yield from both treatments was similar to N at 300 mg-L-1.

Cutting length, stem diameter, and leaf area

There was a harvest date x cultivar interaction for cutting length (Table 2-15).

Differences in cutting length were not significant between cultivars for HI or H3 with 7.7

and 8.2 cm per cutting, respectively. For H5, cuttings harvested from stock plants of

'Purple Small Leaf had 116% longer cuttings than from 'Raspberry Ice' (10.2 vs 8.7 cm)

(LSD=1.1, n=34). When considering only 'Raspberry Ice', cutting length was longer

from H5 (8.7 cm) than H1 (7.9 cm), while H3 was similar to both with 8.4 cm per cutting

(LSD=0.5, n=35). 'Purple Small Leaf had a similar cutting length from HI and H3 (7.5

and 8.0 cm, respectively), both of which were less than H5 (10.1 cm) (LSD=0.9, n=36).

Differences in stem length for either cultivar were not significant amongst treatments.

There was a harvest date x N concentration and a harvest date x cultivar interaction

for stem diameter (Table 2-15). For HI 'Purple Small Leaf had a 16% smaller stem

diameter than 'Raspberry Ice' (0.19 vs 0.23 cm, respectively). Both cultivars had similar

stem diameter at H3 (data not shown). At H5, 'Purple Small Leaf had larger sized stems

(0.24 cm) than 'Raspberry Ice' (0.20 cm) (LSD=0.02, n=34). 'Raspberry Ice' cuttings

from H3 had a stem diameter of 0.24 cm, which was 108% and 115% larger than H1

(0.23) and H5 (0.20), respectively (LSD=0.01, n=35). Alternatively, 'Purple Small Leaf

cuttings had a larger stem diameter at H5 than H3 (0.24 vs 0.21 cm), both of which were

greater than H1 (0.19 cm) (LSD=0.02, n=36). Although statistical differences were









shown, differences in stem diameter of 0.03 cm may not be considered commercially

important for either cultivar. When considering both cultivars, differences in stem

diameter were not significant amongst treatments for any harvest date. When considering

harvest dates within treatments, cuttings harvested from stock plants treated with N at

300 mg-L-1 had a larger stem diameter from H3 (0.23 cm) than either H5 or HI, which

had similar a diameter of 0.20 cm (LSD=0.02, n=24). Differences in stem diameter

amongst harvest dates were not significant for other N concentrations.

There was a harvest date x cultivar and a cultivar x N concentration interaction for

leaf area (Table 2-15). From HI, cuttings of'Raspberry Ice' had a 157% larger leaf area

than 'Purple Small Leaf (76.5 vs 48.5 cm2, respectively) (LSD=4.9, n=36). Similarly, at

H3, 'Raspberry Ice' had a leaf area of 47.1 cm2, which was 115% larger than that of

'Purple Small Leaf (40.8 cm2) (LSD=4.3, n=36). At H5, however, 'Purple Small Leaf

had larger leaf area than 'Raspberry Ice' (31.6 vs 24.1 cm2) (LSD=3.9, n=36). When

considering differences in 'Raspberry Ice' amongst harvest dates, leaf area from H1 (76.5

cm2) was larger than H3, both (47.1 cm2) of which were larger than H5 (24.1 cm2)

(LSD=4.7, n=35). 'Purple Small Leaf had larger leaf area at H5 (10.1 cm2) than at HI or

H3 (7.5 or 8.0 cm2, respectively) (LSD=0.9, n=36).

Differences for both 'Raspberry Ice' and 'Purple Small Leaf were not significant

amongst treatments with a leaf area of 51.0 and 40.3 cm2, respectively (Table 2-16).

When comparing cultivars, stock plants fertigated with N at 100 to 200 mg-L1, had a

larger leaf area with 'Raspberry Ice' than 'Purple Small Leaf. Differences in leaf area

were not significant between cultivars when stock plants were provided N at 300 mg-L1.









Rooting parameters

Percent survival, visual rooting quality, and quantity of primary and lateral roots

There was a harvest date x evaluation week x N concentration interaction for

percent survival (Table 2-17). Main effects for cultivar differences will be presented

separately. Percent survival of'Raspberry Ice' was 93%, while 71% of'Purple Small

Leaf cuttings survived (LSD=5, n=90). Differences in percent survival were not

significant amongst treatments for cuttings planted at H2, H4, or H6 and evaluated 4 or 6

WAP, respectively (data not shown). When considering N concentration, stock plants

fertilized with N at 300 mg-L-1 produced cuttings at H4 that had a higher percent survival

6 WAP than at 4 WAP (86% vs 55%) (LSD=20, n=12), otherwise all other harvest

plantings had similar survival percentages at both 4 and 6 WAP, regardless of N

concentration.

There was a harvest date x cultivar x N concentration and a harvest date x

evaluation week interaction for rooting quality (Table 2-17). When considering both

cultivars, cuttings evaluated 6 WAP had a higher rooting quality than those evaluated 4

WAP from H2, H4, and H6 (data not shown). For both 'Raspberry Ice' and 'Purple Small

Leaf, rooting quality was similar amongst treatments and harvests. One exception was

cuttings of 'Purple Small Leaf planted from stock plants treated with N at 100 mg-L-1,

which had a lower root quality rating than those provided with N at 200 or 300 mg-L-1

(Table 2-18). 'Raspberry Ice' when harvested from N concentrations of 100 mg-L-1,

produced better quality rooted cuttings when planted from H2 and H6, than from H4.

Nitrogen at 200 mg-L-1 produced variable results in rooting quality from each harvest,

with mean ratings of 3.9, 1.6, and 3.0 from H2, H4, and H6, respectively. Similarly,

cuttings planted from H2 had higher rooting quality than H6, which was higher than H4









with N at 300 mg-L-1. While 'Purple Small Leaf cuttings had similar rooting quality from

H4 and H6, rooting quality of cuttings planted from N at 100 mg-L-1 at H2 was higher.

Differences in rooting quality were not significant amongst harvest dates with N

concentrations from 200 to 300 mg-L-1.

There was a harvest date x cultivar x N concentration interaction for number of

primary roots (Table 2-19). For most harvest plantings, both 'Raspberry Ice' and 'Purple

Small Leaf had a similar number of primary roots amongst treatments. However,

'Raspberry Ice' from H2 had more primary roots per cutting when fertilized with N at 200

mg-L-1, than at 300 mg-L-1, and both of these treatments were similar to N at 100 mg-L-1.

Cuttings of 'Purple Small Leaf planted at H6 had 281% more primary roots when stock

plants were fertigated with N at 300 mg-L-1 than at 100 mg-L-1, while both treatments

were similar to N at 200 mg-L-'. For cuttings planted from H2, 'Raspberry Ice' had

264%, 425%, and 237% more primary roots than 'Purple Small Leaf when stock plants

were fertigated with N at 100, 200, and 300 mg-L-1, respectively (data not shown).

'Raspberry Ice' and 'Purple Small Leaf cuttings both had a similar number of primary

roots amongst the three N concentrations, when planted from H4 and H6.

There was a harvest date x cultivar interaction for number of lateral roots.

'Raspberry Ice' had 18.5 lateral roots per cutting for H2, which was at least 506% more

than H4 or H6 (1.4 or 3.7, respectively) (LSD=5.6, n=12). 'Purple Small Leaf had more

lateral roots per cutting when planted from H6 (6.5) than H2 (3.4) or H4 (1.4) (LSD=2.6,

n=18). For cuttings planted from H2, 'Purple Small Leaf had an 82% lower number of

lateral roots than cuttings of 'Raspberry Ice' (data not shown). 'Raspberry Ice' and 'Purple









Small Leaf had a similar number of lateral roots from H4 and H6, with 1.4 and 5.1 lateral

roots per cutting, respectively.

Cutting dry weight and root : shoot ratio

There was a harvest date x evaluation week and a harvest date x cultivar interaction

for cutting total dry weight (Table 2-17). For cuttings planted from H2, those evaluated 6

WAP had greater total dry weight than at 4 WAP (291 vs 231 mg) (LSD=39, n=35). For

H4 and H6, differences in total dry weight were not significant between evaluation weeks

with 219 and 173 mg per cutting, respectively. When comparing cultivars, 'Raspberry

Ice' and 'Purple Small Leaf had similar cutting dry weight when planted from H2 and H6

with 244 and 253 mg per cutting, respectively. For H4, 'Raspberry Ice' cuttings had

greater dry weight than 'Purple Small Leaf (245 vs 211 mg) (LSD=29, n=22). Dry

weight of'Raspberry Ice' cuttings decreased over time with 289, 245, and 163 mg per

cutting from H2, H4, and H6, respectively (LSD=32, n=16). 'Purple Small Leaf

responded similarly over time with 251, 211, and 177 g per cutting from H2, H4, and H6,

respectively (LSD=32, n=45).

There was a harvest date x evaluation week and a harvest date x cultivar interaction

for the cutting root: shoot ratio (Table 2-17). The root : shoot ratio was similar between

'Raspberry Ice' and 'Purple Small Leaf at H2, H4 and H6. 'Raspberry Ice' cuttings had

similar root: shoot ratios for H2 and H6 with 0.24, which was greater than H4 (0.15)

(LSD=0.05, n=16). 'Purple Small Leaf cuttings performed similarly, with H2 (0.24) or

H6 (0.28) being greater than H4 (0.15) (LSD=0.05, n=45). When considering evaluation

weeks, cuttings evaluated from either H4 (0.15) or H6 (0.26) had similar root : shoot

ratios at both 4 and 6 WAP, while cuttings evaluated 4 WAP from H2 had a 42% lower

root: shoot ratio than at 6 WAP (0.17 vs 0.32) (LSD=0.05, n=34).









Stock plant leachate

There was a sampling date x N concentration interaction for leachate pH,

differences in cultivar were not significant. Over time substrate pH increased with N at

100 mg-L-1, while a decrease in pH was observed with N at 200 to 300 mg-L-1 (Figure 2-

4).

The sampling date x N concentration interaction was not significant for leachate

EC, therefore the main effects will be presented separately. Both cultivars had similar

EC values. Over time, values remained within the recommended range of 2.0 to 3.5

mS/cm for producing bougainvillea plants (Figure 2-5). As the concentration of N

applied to stock plants increased, EC also increased. Nitrogen at 300 mg-L-1 had the

highest leachate EC (3.62 mS-cm-1), while 200 mg-L-1 (2.89 mS-cm-1) was higher than

100 mg-L-1 (1.76 mS-cm-1) (LSD=0.32, n=48).

There was a sampling date x N concentration interaction for NO3-N, differences in

cultivar were not significant. In general, leachate values for NO3-N were shown to

increase with removal of plant tissue at cutting harvests when stock plants were fertigated

with N at 200 to 300 mg-L-1 (Figure 2-6). Leachate NO3-N values were excessively high

with N at 300 mg-L-1.

Foliar analysis

There was a harvest date x cultivar x N concentration interaction for foliar Ca and

Mg concentrations. There was a harvest x cultivar interaction for foliar P and K

concentrations. There was a harvest x N concentration interaction for foliar N, while

main effects for cultivar will be presented separately (Table 2-20). 'Raspberry Ice' had

greater foliar N concentration than 'Purple Small Leaf (4.59% vs 4.43%) (LSD=0.14,

n=18). Early in the season, at HI, stock plants fertigated with N at 200 mg-L-1 (4.52%)









had greater foliar N concentration than at 300 mg-L-1 (4.16%), while both concentrations

were similar to N at 100 mg-L-1 (4.30%) (LSD=0.27, n=6). During H3, foliar N

concentration was similar for stock plants fertigated with N at 200 to 300 mg-L-1 (4.94%),

while both were greater than 100 mg-L-1 (4.14%) (LSD=0.33, n=6). Harvest 5 followed a

similar pattern, as the N concentration delivered to stock plants increased, N

concentration in the foliage also increased (data not shown). When considering foliar K

concentrations, cuttings harvested from stock plants treated with N at 100 mg-L-1 had the

highest tissue concentrations (4.16%), while 200 mg-L-1 (3.93%) was greater than 300

mg-L-1 (3.54%) (LSD=0.16, n= 27). 'Purple Small Leaf had a similar K concentration in

foliage to 'Raspberry Ice' during HI and H5, while at H3 'Purple Small Leaf (4.23%) had

a greater K concentration than 'Raspberry Ice' (3.63%) (LSD=0.31, n=9). Phosphorous

levels were similar for all treatments during the second study. During HI, 'Purple Small

Leaf cuttings had a greater foliar P concentration than 'Raspberry Ice' (0.52% vs 0.42%)

(LSD=0.05, n=9). The third harvest followed an opposite trend, with P being greater in

'Raspberry Ice' (0.32%), than 'Purple Small Leaf (0.29%) (LSD=0.02, n=9). Differences

in cultivar responses were not significant at H5, with 0.26% P measured in tissue. Over

time, foliar P and K concentrations decreased for both cultivars, although values were

still within the acceptable range of 0.2% to 0.8% for P, and 3.0 to 5.5% for K (Whipker et

al., 2003).

When considering foliar Ca levels observed for 'Raspberry Ice' and 'Purple Small

Leaf, concentrations were generally similar amongst N concentrations applied. During

H3 however, decreased Ca concentrations were observed as N concentration increased

(Table 2-21). As the season progressed, 'Raspberry Ice' exhibited a decline in foliar Ca.









Although the characteristic leaf distortions and chlorosis commonly associated with Ca

deficiency were not observed, resultant slow growth due to a minor calcium deficiency

may have contributed to the reduction in cutting quantity harvested later in the season

(Whipker et al., 2003). Magnesium levels were shown to be above the recommended

range of 0.2% to 0.8% during HI for both cultivars (Table 2-21). Decreased foliar Mg

was observed as the season progressed for both cultivars, however values remained

within the recommended range.

Discussion

During the first experiment, 'Purple Small Leaf produced more cuttings than

'Raspberry Ice'. Although cutting length, stem diameter, and leaf area were similar

between cultivars, percent survival, rooting quality, number of lateral roots, total cutting

dry weight, and root: shoot ratios for planted cuttings were shown to be greater with

'Raspberry Ice' than 'Purple Small Leaf.

When considering both cultivars, stock plants fertigated with N concentrations of

200 to 300 mg-L-1, produced greater quantities of cuttings, when compared to 100 mg-L-1.

At times, all treatments produced similar numbers of cuttings, yet throughout the study N

at 300 mg-L-1, more consistently produced the largest yield (Table 2-2). Additionally,

when comparing consistency of harvest dates, stock plants fertigated with N at 100 or

300 mg-L-1 produced similar numbers of cuttings at each harvest, while results from N at

200 mg-L-1 varied.

'Purple Small Leaf produced cuttings with a shorter stem length than 'Raspberry

Ice', however the 0.3 cm difference may not be considered commercially important.

When considering N concentration, cutting length was generally similar amongst

treatments, however at the third harvest stock plants fertigated with N at 100 to 200









mg-L-1 produced longer cuttings than at 300 mg-L-1. Cutting leaf area was generally

similar for all treatments, although 'Raspberry Ice' showed a reduction in area as N

concentration increased. 'Purple Small Leaf had a similar leaf area for all N

concentrations provided. As the N concentration increased later in the harvesting season,

a slight trend was shown with both cultivars toward shorter cuttings with decreased leaf

area.

Percent survival for cuttings planted from H2 was higher with cuttings planted

from stock plants treated with N at 300 mg-L-1 than from concentrations of 100 to 200

mg-L-1, otherwise all treatments had similar survival. When comparing harvest dates,

cuttings planted from H6 had a considerable reduction in survival as compared to the

other two plantings. Improved survivability with N at 200 mg-L-1 than 300 mg-L-1,

occurred with cuttings planted from H6, therefore growers may want to consider reducing

fertilizer concentrations as the growing season progresses into the cool season. Rooting

quality of 'Purple Small Leaf cuttings was improved with N at 300 mg-L-1, as compared

to lower concentrations early in the study, yet as the season progressed treatment

differences were not significant. 'Raspberry Ice' cutting performance was generally not

influenced by treatment early in the season, yet had better rooting quality when planted

from H6 when N was applied at 200 mg-L-1, as compared to 100 or 300 mg-L-1. When

comparing primary root number, all N concentrations produced a similar quantity. In

general both cultivars had a reduction in primary roots as the season progressed; this

trend seems to coincide with the decreases in visual rooting quality shown during the

same time period. For 'Raspberry Ice', this reduction in rooting quality at H6 may be

related to the reduction in both stem length and leaf area seen with increased









concentrations of nitrogen applied to stock plants later in the season. Lateral root number

was generally improved with N concentrations of 200 to 300 mg-L-1 when applied to

stock plants of 'Raspberry Ice', and with 300 mg-L-1 applied to 'Purple Small Leaf.

Differences in total cutting dry weight and the root: shoot ratio seem to coincide

with other rooting parameters previously discussed, as 'Purple Small Leaf was shown to

have increased total cutting dry weight and an improved root: shoot ratio with N at 300

mg-L-1 as compared to the lower concentrations applied to stock plants. Although

'Raspberry Ice' was not significant amongst treatments for the root : shoot ratio, total

cutting dry weight was greater from N at 200 to 300 mg-L-1 than at 100 mg-L-1.

When comparing all parameters, stock plants of 'Raspberry Ice' and 'Purple Small

Leaf were shown to achieve the greatest benefits in yield and subsequent rooting

performance when stock plants were fertigated with N at 200 to 300 mg-L-1 during Expt.

1 (warm season). Results were similar to studies conducted during the 2005 spring

growing season, in which the recommended concentration was 200 mg-L-1 (Cerveny et

al., 2005).

During the second experiment, 'Purple Small Leaf produced more cuttings than

'Raspberry Ice'. When comparing treatments, 'Raspberry Ice' produced the greatest

quantity of cuttings when stock plants were fertigated with N at 300 mg-L-1 as compared

to 100 to 200 mg-L-1. Alternatively, 'Purple Small Leaf stock plants were shown to have

optimum yield with N at 200 mg-L-1 as compared to 100 or 300 mg-L-1, suggesting no

benefit to the increased concentration.

Cutting length was generally similar for both cultivars, except 'Purple Small Leaf

produced longer cuttings than 'Raspberry Ice' during H5. 'Raspberry Ice' generally had a









larger leaf area than 'Purple Small Leaf with lower concentrations of N applied, yet

differences were not significant as concentrations increased to 300 mg-L-1. When

comparing harvests, leaf area tended to be greater with 'Raspberry Ice' than 'Purple Small

Leaf during earlier harvests, although at H5 'Purple Small Leaf had a larger leaf area

than 'Raspberry Ice'. As the season progressed into the cooler months, 'Raspberry Ice'

stock plants produced fewer cuttings with shorter lengths and less leaf area; this was

shown to a lesser degree with 'Purple Small Leaf. This change in performance might

suggest a seasonal effect, although a similar trend was observed to a lesser degree during

summer. Because bougainvillea flowers under natural short days, flower initiation may

have inhibited vegetative growth (Chakraverty, 1970). Another possibility for the

observed changes in cutting yield may be due to a natural reduction in vigor associated

with 18 to 20 weeks in production. In commercial floriculture, stock plants of other

species are often replanted several times per year to avoid reductions in vigor (Carlos

Villalobos, General Manager, Ball Linda Vista facility, Costa Rica, personal

communication). Alternatively, the foliar analysis data which showed slight Ca and Mg

deficiencies, coupled with reduced pH values observed in stock plant leachate may

indicate reductions in vigor are related to elemental deficiencies associated with low pH.

Percent survival was generally not influenced by N concentration applied to stock

plants and 'Raspberry Ice' tended to be similar in percentage survival to 'Purple Small

Leaf for most harvest plantings and evaluation dates. Visual rooting quality was also

generally similar amongst N concentration provided to stock plants, except for 'Purple

Small Leaf later in the season, in which higher rates of N at 200 to 300 mg-L-1 provided

to stock plants had greater rooting quality than at 100 mg-L-1. Throughout the second









study N fertilization provided to 'Purple Small Leaf at 200 to 300 mg-L-1 produced

similar results when planted from all harvests. 'Raspberry Ice' tended to have better

rooting earlier in the study for all treatments, but was generally similar amongst all N

concentrations.

For the N concentrations investigated, 200 and 300 mg-L-1 produced the highest

quantity of primary roots for 'Raspberry Ice' and 'Purple Small Leaf, respectively. For

lateral roots, differences in treatment were not significant, yet 'Raspberry Ice' was shown

to have improved lateral rooting earlier in the season, while 'Purple Small Leaf

performed better as the season progressed. 'Raspberry Ice' was greater than 'Purple Small

Leaf when considering number of lateral roots when cuttings were planted from H2,

although H4 and H6 produced similar results between cultivars.

'Raspberry Ice' and 'Purple Small Leaf generally had similar total cutting dry

weight values throughout the study with few exceptions. Total cutting dry weight of

'Raspberry Ice' decreased with each harvest planting, while 'Purple Small Leaf values

were constant over time. This data further illustrates differences observed with other

parameters previously mentioned. The root: shoot ratios were similar for 'Raspberry Ice'

and 'Purple Small Leaf cuttings, and both followed a similar trend with harvest number.

Cutting root : shoot ratios were generally similar with each harvest, however a reduction

was seen when cuttings were planted from H4.

When considering both studies, cuttings evaluated 4 WAP tended to have similar

survival to those evaluated 6 WAP, this suggests that although rooting may not be at the

desired level, no benefit was shown in allowing cuttings to remain under mist for six

weeks. All other parameters that showed improvement 6 WAP as compared to 4 WAP,









such as rooting quality, root number, and dry weight, all suggest growth with time and

would not be considered the result of prolonged misting. Growers will want to consider

removing cuttings from the propagation environment when the majority of cuttings have

begun rooting to encourage better establishment of rooted liners. Length of time

necessary to propagate bougainvillea cuttings varies, so growers will want to experiment

with each cultivar to determine the ideal harvesting window.

Conclusion

During the summer study stock plants of bougainvillea were shown to benefit from

N at 300 mg-L-1. The trend during the second study suggests that 300 mg-L-1 may be

more appropriate earlier in the growing season, but as the season progresses fertilization

program should be reduced to 200 mg-L-1. Stock plant leachate pH, EC, and NO3-N

should be monitored regularly to assure optimum root zone management. Bougainvillea

glabra 'Raspberry Ice' and 'Purple Small Leaf were also shown to survive similarly at 4

and 6 WAP under Florida conditions. Growers should compare the risk of extended

propagation time to the benefits achieved in rooting performance for their specific

growing environments. Since the response of 'Purple Small Leaf and 'Raspberry Ice'

varied over time and between each other, further studies should be conducted on other

cultivars of bougainvillea as well as other tropical species to determine optimum

concentrations of N for a successful stock plant management program.









Table 2-1. Temperature and relative humidity data (maximum, minimum, and mean) as
recorded during simulated and actual shipping (excision to planting) of
cuttings from Harvests (H) 2, 4, and 6 of Expts. 1 and 2, respectively.
Temperature (C) Relative Humidity (%)
Expt 1 Max. Min. Mean Max. Min. Mean
H2 34.4 13.3 22.4 94.5 46.4 81.4
H4 36.1 20.6 26.4 82.5 56.2 69.1
H6 38.8 22.9 26.5 91.7 36.3 72.3
Avg 36.4 18.9 25.1 89.6 46.3 74.3

Temperature Relative Humidity
Expt 2 Max. Min. Mean Max. Min. Mean
H2 38.8 21.0 25.8 82.1 30.4 65.1
H4 26.0 3.3 17.3 96.7 26.6 63.7
H6 25.6 3.3 13.9 88.0 30.4 72.7
Avg 30.1 9.2 19.0 88.9 29.1 67.2


Table 2-2. Influence of harvest date, cultivar, and nitrogen (N) concentration on quantity
of sub-terminal bougainvillea stem cuttings harvested during the warm season
(Expt. 1).
N Concn Cutting quantity
(mg-L-1) HI H2 H3 H4 H5 H6 LSD
100 13.5 16.1 16 15.9 15.6 9.5 NS
200 14 19 17.6 20 25.5 11.4 7
300 13.2 20.1 22.1 20.8 23.9 18.1 NS
Significance NS NS *** NS *
LSD 3.6 6.6 6.9

Source of variance df Pr > F
Rep (R) 11 0.0061
Harvest date (H) 5 0.0001
Cultivar (C) 1 0.0001
HxC 5 0.0001
N concn (T) 2 0.0001
HxT 10 0.0495
C xT 2 NS
HxCxT 10 NS
NS, *, *** Nonsignificant or significant at P < 0.05 or 0.001, respectively.









Table 2-3. Significance levels for effects of harvest date, cultivar, and nitrogen (N)
concentration on length and leaf area of cuttings harvested from stock plants
of bougainvillea (Expt 1).
Source df Cutting length Leaf area
Rep (R) 11 NS NS
Harvest date (H) 2 0.0001 0.0001
Cultivar (C) 1 0.0254 0.0001
HxC 2 NS NS
N concn (T) 2 NS 0.0018
HxT 4 0.0454 NS
C xT 2 NS 0.0063
HxCxT 4 NS NS
NSNonsignificant at P < 0.05.


Table 2-4. Harvest date (H) by nitrogen (N) concentration interaction for length of sub-
terminal bougainvillea stem cuttings from harvested during the warm season
(Expt 1).
Cutting length (cm)
N Concn
(mg-L-1) H1 H3 H5 LSD
100 7.3 6.7 7.5 0.6
200 6.7 6.7 7.3 0.5
300 6.9 6 7.6 *** 0.6
Significance NS NS
LSD 0.5
NS, ** Nonsignificant or significant at P < 0.05 or 0.001, respectively.









Table 2-5. Significance levels for effects of harvest date, evaluation date, cultivar, and
nitrogen concentration provided to stock plants on percent survival, visual
rooting quality (l=poor; 5=best), total dry weight (shoot weight + root
weight), and root : shoot ratio of sub-terminal bougainvillea stem cuttings
planted during the warm season (Expt. 1).


Source
Rep (R)
Harvest date (H)
Evaluation date (W)
HxW
Cultivar (C)
HxC
WxC
HxWxC
N Concn (T)
HxT
WxT
HxWxT
CxT
HxCxT
WxCxT
HxWxCxT
NSNonsignificant at P


df
5
2
1
2
1
2
1
2
2
4
2
4
2
4
2
4
< 0.05.


Percent
survival
NS
0.0001
NS
NS
0.0001
NS
NS
NS
NS
0.0051
NS
NS
0.0227
NS
NS
NS


Rooting
quality
NS
0.0001
0.0185
NS
0.0001
0.0001
NS
NS
NS
0.0005
NS
NS
0.0003
0.0482
NS
NS


Total dry
weight
0.0248
0.0001
0.0001
0.0001
0.0001
0.0041
NS
0.0001
0.0001
0.0001
NS
NS
NS
0.006
NS
NS


Root: shoot
ratio
NS
0.0001
0.0001
0.0019
0.0001
0.0001
0.0015
0.0482
NS
0.0011
NS
0.0314
0.0096
NS
NS
NS


Table 2-6. Harvest date (H) by nitrogen (N) concentration interaction for percent
survival of sub-terminal bougainvillea stem cuttings planted from stock plants
treated with N at 100, 200, or 300 mg-L-1 during the warm season (Expt. 1).
Percent survival (%)
N Concn
(mg-L-1) H2 H4 H6 LSD
100 75 77 58 15
200 76 69 63 NS
300 90 80 49 *** 14
Significance NS NS
LSD 12
Ns, *, *** Nonsignificant or significant at P < 0.05 or 0.001,
respectively.









Table 2-7. Harvest date (H) by cultivar by N concentration interaction for visual rooting
quality (l=poor; 5=best) of sub-terminal bougainvillea stem cuttings planted
during the warm season (Expt. 1).
Rooting quality
N Concn Raspberry Ice Purple Small Leaf
(mg-L-1) H2 H4 H6 H2 H4 H6
100 3.6 2.8 1.5 1.8 1.3 1.6
200 3.9 2.8 2 1.5 1.1 1.3
300 3.5 2.2 1.3 2.9 1.4 1.1
Significance NS NS NS NS
LSD 0.5 0.8
Ns, Nonsignificant or significant at P < 0.05, respectively.


Table 2-8. Significance levels for effects of harvest date, cultivar, and nitrogen
concentration provided to stock plants on number of primary and lateral roots
on sub-terminal bougainvillea stem cuttings planted during the warm season
(Expt. 1).
Source df Primary roots Lateral roots
Rep (R) 5 NS NS
Harvest date (H) 2 0.0001 0.0001
Cultivar (C) 1 0.0001 0.0001
HxC 2 0.0001 0.0001
N Concn (T) 2 NS 0.0129
HxT 4 NS 0.0094
C xT 2 0.0343 0.0003
HxCxT 4 NS 0.0278
NSNonsignificant at P < 0.05.


Table 2-9. Harvest date by cultivar by N concentration interaction for total cutting dry
weight (shoot weight + root weight) of sub-terminal bougainvillea stem
cuttings planted during the warm season (Expt. 1).
Total cutting dry weight (mg)
N Concn Raspberry Ice Purple Small Leaf
(mg-L-1) H2 H4 H6 H2 H4 H6
100 204 152 216 116 98 145
200 253 164 230 138 101 139
300 246 179 222 184 105 111
Significance NS NS *** NS *
LSD 36 25 23
Ns,*, *** Nonsignificant or significant at P < 0.05 or 0.001, respectively.










Table 2-10. Significance levels for effects of harvest date, cultivar, and nitrogen (N)
concentration on foliar concentration of macronutrients N, phosphorous (P),
potassium (K), calcium (Ca), and magnesium (Mg) of sub-terminal
bougainvillea stem cuttings harvested during the warm season (Expt. 1).
Source df N P K Ca Mg
Rep 2 NS NS NS NS NS
Harvest date (H) 2 NS 0.0001 0.0001 0.0001 0.0001
Cultivar (C) 1 NS 0.0001 0.0001 0.0001 NS
HxC 2 0.0396 0.0038 0.0114 0.0001 0.0001
N concn (T) 2 0.0001 0.0368 0.0001 0.0001 0.0001
HxT 4 0.0213 0.0028 NS 0.0001 0.0002
C x T 2 NS 0.0009 0.0040 NS 0.0001
HxCxT 4 NS NS NS NS NS
NSNonsignificant at P < 0.05.



Table 2-11. Harvest date (H) by cultivar interaction for foliar nutrient concentration of
macronutrients nitrogen (N), phosphorous (P), potassium (K), calcium (Ca),
and magnesium (Mg) of sub-terminal bougainvillea stem cuttings harvested
during the warm season (Expt. 1).
Foliar concn (%)
Cultivar Harvest N P K Ca Mg
Raspberry Ice HI 4.63 0.40 3.92 0.96 0.62
Purple Small Leaf 4.57 0.37 4.65 1.16 0.52
NS *** *** **
Raspberry Ice H3 5.02 0.33 3.94 0.82 0.55
Purple Small Leaf 4.59 0.33 4.31 1.14 0.60
NS *** NS
Raspberry Ice H5 4.80 0.33 3.75 0.83 0.51
Purple Small Leaf 4.87 0.29 4.21 0.87 0.51
NS *** *** NS NS
Ns, Nonsignificant or significant at P < 0.05, 0.01, or 0.001, respectively.










Table 2-12. Harvest date (H) by nitrogen (N) concentration interaction for foliar nutrient
concentrations of macronutrients N, phosphorous (P), calcium (Ca), and
magnesium (Mg) of sub-terminal bougainvillea stem cuttings harvested during
the warm season (Expt. 1)..


N concn
Harvest (mg-L-1)
H1 100
200
300


100
200
300

100
200
300


Foliar concn (%)


N
4.52
4.68
4.60
NS
4.56
4.81
5.05
NS
4.36
4.84
5.31
***


P
0.39
0.38
0.40
NS
0.31
0.32
0.35
*
0.31
0.32
0.30
NS


Ca
1.12
1.11
0.96
**
1.11
1.06
0.77

1.13
0.85
0.57
***


Mg
0.61
0.59
0.51
*
0.65
0.62
0.46

0.63
0.52
0.39
***


NS, **' Nonsignificant or significant at P < 0.05, 0.01, or 0.001,
respectively.
z Any two means within a column not followed by the same letter are
statistically different using LSD at P < 0.05.


Table 2-13. Cultivar by nitrogen (N) concentration interaction for foliar nutrient
concentration of macronutrients phosphorous (P), potassium (K), and
magnesium (Mg) of sub-terminal bougainvillea stem cuttings harvested during
the warm season (Expt. 1).
Foliar concn (%)
N concn (mg-L-1) P K Mg
Raspberry Ice 100 0.35 4.00 a 0.62 a
200 0.36 3.94 a 0.55 b
300 0.35 3.66 b 0.51 c
NS *** ***
Purple Small
Leaf 100 0.33 4.62 a 0.52 b
200 0.32 4.22 b 0.60 a
300 0.35 4.32 b 0.51 b
NS ** ***
Ns, Nonsignificant or significant at P < 0.05, 0.01, or 0.001, respectively.
z Any two means within a column not followed by the same letter are statistically
different using LSD at P < 0.05.









Table 2-14. Influence of harvest date, cultivar, and nitrogen (N) concentration on
quantity of sub-terminal bougainvillea stem cuttings harvested during the cool
season (Expt. 2).
N concn Cutting quantity
(mg-L-1) Raspberry Ice Purple Small Leaf LSD
100 4.9 13.9 *** 2
200 4.9 17.5 *** 2.3
300 6.6 15.8 *** 1.8
Significance ** *
LSD 1 2.2

Source of variance df Pr > F
Rep (R) 11 0.0241
Harvest date (H) 5 0.0001
Cultivar (C) 1 0.0001
HxC 5 0.0001
N concn (T) 2 0.0037
HxT 10 NS
C x T 2 0.006
HxCxT 10 NS
NS, ,, *** Nonsignificant or significant at P < 0.05, 0.01, 0.001, respectively.



Table 2-15. Significance levels for effects of harvest date, cultivar, and nitrogen (N)
concentration on length, stem diameter, and leaf area of sub-terminal
bougainvillea stem cuttings harvested during the cool season (Expt. 2).

Source df Cutting length Stem diameter Leaf area
Rep (R) 11 NS NS 0.0051
Harvest date (H) 2 0.0001 0.0044 0.0001
Cultivar (C) 1 NS 0.0221 0.0001
HxC 2 0.0012 0.0001 0.0001
N concn (T) 2 NS NS NS
HxT 4 NS 0.0283 NS
C xT 2 NS NS 0.0002
HxCxT 4 NS NS NS
NSNonsignificant at P < 0.05.










Table 2-16. Cultivar by nitrogen (N) concentration interaction for leaf area of sub-
terminal bougainvillea stem cuttings harvested during the cool season (Expt.
2).
N concn Leaf area (cm2)
(mg-L-1) Raspberry Ice Purple Small Leaf LSD
100 50.6 37.6 9.1
200 53.5 40.1 9.3
300 45.8 43.2 NS
Significance NS NS

Ns, Nonsignificant or significant at P < 0.05, respectively.



Table 2-17. Significance levels for effects of harvest date, evaluation date, cultivar, and
nitrogen concentration provided to stock plants on percent survival, visual
rooting quality (l=poor; 5=best), total dry weight (shoot weight + root
weight), and root : shoot ratio of sub-terminal bougainvillea stem cuttings
planted during the cool season (Expt. 2).


Source df
Rep (R) 5
Harvest date (H) 2
Evaluation date (W) 1
HxW 2
Cultivar (C) 1
HxC 2
WxC 1
HxWxC 2
N Concn (T) 2
HxT 4
WxT 2
HxWxT 4
CxT 2
HxCxT 4
WxCxT 2
HxWxCxT 4
NSNonsignificant at P < 0.05.


Percent
survival
NS
0.0001
NS
0.0038
0.0001
NS
NS
NS
NS
NS
NS
0.0480
NS
NS
NS
NS


Rooting
quality
NS
0.0001
0.0001
0.0032
0.0732
0.0001
NS
NS
NS
NS
NS
NS
NS
0.0024
NS
NS


Total dry
weight
NS
0.0001
0.0083
0.0001
0.0024
0.0001
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS


Root: shoot
ratio
NS
0.0001
0.0001
0.0001
NS
0.0032
NS
NS
NS
NS
NS
NS
NS
NS
NS
NS










Table 2-18. Harvest date (H) by cultivar by nitrogen (N) concentration interaction for
visual rooting quality (1-5; l=poor, 5=best) of sub-terminal bougainvillea
stem cuttings planted during the cool season (Expt. 2).
Rooting quality
N concn Raspberry Ice Purple Small Leaf
(mg-L-1) H2 H4 H6 H2 H4 H6
100 3.6 1.8 3 3.2 2.4 2.4
200 3.9 1.7 3 2.6 2.3 3
300 4.1 1.6 2.7 2.5 2.5 3.1
Significance NS NS NS NS NS *
LSD 0.6
Ns, Nonsignificant or significant at P < 0.05, respectively.


Table 2-19. Influence of harvest date (H), cultivar, and nitrogen (N) concentration on
quantity of primary roots of sub-terminal bougainvillea stem cuttings planted
during the cool season (Expt. 2).
Number of primary roots
N concn Raspberry Ice Purple Small Leaf
(mg-L-1) H2 H4 H6 H2 H4 H6
100 8.8 0.5 3.2 3.3 3.1 2.1
200 10.2 0.5 6.2 2.4 4.6 3.6
300 6.2 1 2.7 2.6 1.7 5.9
Significance NS NS NS NS *
LSD 2.6 2.8

Source of variance df Pr> F
Rep (R) 5 NS
Harvest date
(H) 2 0.0001
Cultivar (C) 1 0.0259
HxC 2 0.0001
N Concn (T) 2 0.0483
HxT 4 NS
C xT 2 NS
HxCxT 4 0.0059
NS, *Nonsignificant or significant at P < 0.05.










Table 2-20. Significance levels for effects of harvest date, cultivar, and nitrogen (N)
concentration on foliar concentration of macronutrients N, phosphorous (P),
potassium (K), calcium (Ca), and magnesium (Mg) of sub-terminal
bougainvillea stem cuttings harvested during the cool season (Expt. 2).
Source df N P K Ca Mg
Rep 2 NS NS NS NS NS
Harvest date (H) 2 0.0006 0.0001 0.0001 0.0001 0.0001
Cultivar (C) 1 0.0213 0.0048 0.0041 0.0001 0.0001
HxC 2 NS 0.0001 0.0003 NS 0.0036
N concn (N) 2 0.0001 NS 0.0001 NS 0.0013
HxT 4 0.0001 NS NS NS NS
C xT 2 NS NS NS 0.0024 0.0052
HxCxT 4 NS NS NS 0.0046 0.0330
NSNonsignificant at P < 0.05.


Table 2-21. Harvest date (H) by cultivar by nitrogen (N) concentration interaction for
foliar nutrient concentration of calcium (Ca) and magnesium (Mg) in sub-
terminal bougainvillea cuttings harvested from stock plants fertigated with N
at concentrations of 100, 200, or 300 mg-L-1 during the cool season (Expt. 2).
Ca (%)
N concn Raspberry Ice Purple Small Leaf
(mg-L-1) HI H3 H5 HI H3 H5
100 1.40 1.04 a 1.15 1.64 b 1.25 1.03
200 1.44 0.79 b 0.57 1.70 a 1.28 1.38
300 1.56 0.55 c 0.76 1.73 a 1.25 1.01
NS ** NS ** NS NS

Mg (%)
N concn Raspberry Ice Purple Small Leaf
(mg-L-1) H1 H3 H5 H1 H3 H5
100 1.07 0.73 a 0.75 1.03 0.81 0.70
200 1.07 0.54 b 0.55 1.07 0.76 0.77
300 1.02 0.43 c 0.45 1.05 0.73 0.62
NS ** NS NS NS NS
NS, Nonsignificant or significant at P < 0.05, 0.01, or 0.001, respectively.
z Any two means within a column not followed by the same letter are statistically
different using LSD at P < 0.05.







52



7


5
4
3

2
1


0 2 4 6 8 10 12 14 16 18 20 22 24
Weeks after potting

--- 100 -- 200 300 A Harvest date


Figure 2-1. Nitrogen (N) concentration x sampling date interaction for leachate pH
collected from bougainvillea stock plants fertilized with N at 100, 200, or 300
mg-L-1 during the warm season (Expt. 1). Each data point represents mean of
three samples. Cutting harvests are indicated with a triangle along x-axis.


4.5
4
3.5

2.5
9 2
O 1.5
1
0.5
0 A A A A A A
0 2 4 6 8 10 12 14 16 18 20 22 24
Weeks after planting

-*-EC A Harvest date


Figure 2-2. Main effects for leachate electrical conductivity (EC) over time, as recorded
from bougainvillea stock plants during the warm season (Expt. 1). Each data
point represents the mean of three samples. Cutting harvests are indicated by
a triangle along the x-axis.











900
800
700
600
500
z 400
300
S200
100

0 2 4 6 8 10 12 14 16 18 20 22 24
Weeks after planting

-- Raspberry Ice -m- Purple Small Leaf A Harvest date


Figure 2-3. Nitrogen (N) concentration x sampling date interaction for nitrate nitrogen
(NO3-N) values recorded from stock plant leachate of 'Raspberry Ice' and
'Purple Small Leaf bougainvilleas (Expt. 1). Each data point represents the
mean of three samples. Cutting harvests are indicated by a triangle along the
x-axis.


7
6
5
4
3
2
1
0 A A A A A A
0 2 4 6 8 10 12 14 16 18 20 22
Weeks after planting

--100 -0 200 300 A Harvest date


Figure 2-4. Nitrogen (N) concentration by sampling date interaction for leachate pH
collected from bougainvillea stock plants fertilized with N at 100, 200 or 300
mg-L-1 during the cool season (Expt. 2). Each data point represents the mean
of three samples. Cutting harvest dates are indicated by a triangle along the x-
axis.







54



3.5


2.5
E
U 2

1.5
"' 1

0.5


0 2 4 6 8 10 12 14 16 18 20 22
Weeks after planting

*-EC A Harvest date


Figure 2-5. Main effects for leachate electrical conductivity (EC) over time as recorded
from bougainvillea stock plants during the cool season (Expt. 2). Each data
point represents the mean of three samples. Cutting harvests are indicated by
a triangle along the x-axis.



1800
1600
1400
Li 1200
S1000
2 800
6 600
400

200

0 2 4 6 8 10 12 14 16 18 20 22
Weeks after planting

---100 --w200 -- 300 A Harvest date


Figure 2-6. Nitrogen (N) concentration by sampling date interaction for leachate nitrate
nitrogen (NO3-N) values recorded from stock plants of bougainvillea fertilized
with N at 100, 200 or 300 mg-L-1 during the cool season (Expt. 2). Each data
point represents the mean of three samples. Cutting harvests are indicated by
a triangle along the x-axis.














CHAPTER 3
LATERAL BRANCHING AGENTS

Introduction

With the ability to withstand high humidity levels and extremely warm

temperatures in southern climates and the versatility to be used as annuals in the northern

landscapes, tropical perennials offering vibrant color choices and interesting foliar

textures for U.S. consumers (Palmer, 2000). Oleander (Nerium oleander L.) 'Dwarf

Salmon' and Tecoma [Tecoma stans (L.) Juss.] 'Esperanza' are two highly desirable

tropical perennials because of their outstanding flowering characteristics and superior

garden performance.

Oleander is a multipurpose tropical perennial, hardy to USDA Hardiness Zone 9,

that can be used as a foundation plant, hedge, or as a featured specimen in a container

garden. Once established, oleander is able to withstand poor soil conditions and is

reportedly salt tolerant (Walker and Hanly, 1996). It requires a sunny location for

northern gardeners, who could use it as a potted plant to be taken indoors during winter

months (McCulla, 1973). Tecoma, characterized by its trumpet-shaped yellow flowers

was recognized as a 2005 "Plant of the Year" by the Florida Nursery, Growers and

Landscape Association (FNGLA, 2005). Used as a deciduous shrub or tree and hardy to

USDA Hardiness Zone 10, tecoma grows to 7.6 m (25 ft) at maturity, and is equally

attractive as a containerized specimen plant.

While demand is increasing for both species, shortages in rooted cutting

availability exist amongst shipments to growers. This, coupled with low quality rooted









liners supplied by domestic propagation firms has become a concern for growers wishing

to produce oleander and tecoma for sale. As a means of increasing cutting number the

effects of ethephon (Florel) [3.9% (2-chloroethyl) phosphonic acid] (Monteray Lawn and

Garden Products, Inc., Fresno, Calif.) and benzyladenine (BA) on stock plant cutting

quantity has been proven beneficial for promotion of axillary shoot development in some

floriculture crops such as: chrysanthemum (Chiy ,iwniheniun morifolium); geranium

(Pelargonium x hortorum L'Her.); poinsettia (Euphorbiapulcharrima Willd. ex Klotsch);

scaevola (Scaevola aemula R. Br.); and vinca vine (Vinca major L.) (Gibson and

Whipker, 2004; Konjoian, 1994; Witaszek and Mynett, 1989). Because oleander and

tecoma display limited branching in containers, they are ideal candidates for lateral

branching chemical sprays to increase cutting quantity and quality. The objective of the

experiment was to determine the efficacy of lateral branching treatments on stock plants

of Nerium oleander 'Dwarf Salmon' and Tecoma stans 'Esperanza'and to determine the

most effective concentrations of Fresco (N-[phenylmethyl]-lH-purine 6-amine

+gibberellins A4A7: 1.8%)(Fine Agrochemicals Ltd., Worchester, United Kingdom),

FAL-457 (N-[phenylmethyl]-lH-purine 6-amine: 2.0%)(Fine Agrochemicals Ltd.), and

Florel for increasing harvest quantity, quality, and subsequent rooting performance of

cuttings.

Materials and Methods

Experiment 1

Rooted liners of tecoma (Tecoma stans 'Esperanza') and oleander (Nerium

oleander 'Dwarf Salmon') were planted, 2-per pot, into 16d x 18.5h cm round plastic

containers on 18 May 2005. The root substrate used was Fafard 4-P (Fafard, Anderson,

S.C.), which contained: 4 sphagnum peat: 2 pine bark: 2 vermiculite: 1 perlite (v/v).









Slow release fertilizer (19N-2.6P-9.9K) was incorporated into the stock plant rooting

substrate at a rate of 3.21 g-m-3 (1.33 oz/ft3) prior to planting. Plants were then fertigated

once-weekly using 14N-1.7P-11.6K with N at 200 mg-L-1. Plants were established for 4

weeks, until roots reached the bottom of the pot, and then given a hard pinch on 6 July.

Stock plants were sprayed, using a volume of 204 mL-m3, with a treatment solution

consisting of Fresco or FAL457 at 125, 250, or 500 mg-L-1 or Florel at 250, 500, or 1000

mg-L-1. All treatments were mixed with water containing 0.66 mg-L-1 polyether modified

polysiloxane (Capsil) (The Scotts Company, Marysville, Ohio) spray adjuvant. A control

treatment was included, which was sprayed with water and adjuvant only. Treatments

were applied 7 July, 22 July, 4 Aug., and 24 Aug., and cuttings were harvested 4 and 8

weeks after treatments were initiated, on 3 Aug. and 30 Aug., respectively. Stock plants

were assigned to a randomized complete block design (RCBD) with 10 treatments and

five single pot replications of each treatment.

Experiment 2

The second experiment was repeated in the same manner as the first, with the

following exceptions: Stock plants were transplanted on 5 Oct. and given a hard pinch on

31 Oct. Lateral branching agents with nine single pot replications of the 10 treatments

were applied 1 Nov., 15 Nov., 29 Nov., and 13 Dec. Cuttings were harvested on 28 Nov.

and 28 Dec. 2005.

Harvesting procedure

At each harvest, all 2-node tip cuttings were removed from stock plants of oleander

and tecoma. Cutting quantity was recorded along with stem length and diameter (cm) of

three random cuttings per plant. Cuttings were then dipped (0.5 cm for 3 s) in a 1,500

mgL-1 solution containing indolebutyric acid (IBA) 1%: napthaleneacetic acid (NAA)









0.5% and then inserted in a substrate containing 4 perlite: 1 vermiculite (v/v). Cuttings

were assigned to a completely randomized design (CRD) and propagated under mist (5 s

every 20 min). Each species was randomized separately and was assigned, when

numbers were available, with 6 sub-samples and 4 replications of each treatment.

Cutting evaluations

Cuttings were evaluated 5 weeks after planting for percent survival and rooting

quality. Root quality rankings were assigned on a 1 to 5 scale, relative to each species,

with a general ranking system as follows: 1=0 roots with some callus formation; 2=

minimal rooting, 1 or more primary roots around cutting base; 3=5 or more primary roots

with some elongation; 4=uniform root growth and development around cutting base with

primary root elongation and some lateral root growth; 5= best rooting performance

uniform primary and lateral root development with well defined root-ball.

All data were subject to analysis of variance using generalized linear model

procedure in SAS (SAS institute, Cary, N.C.) and means separated using LSD with

P<0.05.

Results

Experiment 1

Harvest yield, cutting length, and stem diameter

The species x treatment interaction was significant for quantity and length of

cuttings, while the harvest date x species x treatment interaction was only significant for

stem diameter (Table 3-1). The harvest date x treatment interaction was significant for

cutting length. The main effects of harvest date will be presented separately for cutting

quantity.












Table 3-1. Significance levels for effects of harvest date, cultivar, and branching
treatment on quantity, length, and stem diameter of 2-node stem cuttings
harvested from stock plants of oleander and tecoma during the cool season
(Expt. 1).
Source df Quantity Length Diameter
Rep 4 NS NS NS
Harvest date (H) 1 0.0020 0.0001 0.0026
Cultivar (C) 1 0.0001 0.0001 0.0001
HxC 1 NS NS NS
Branching treatment (T) 9 0.0001 0.0001 0.0001
HxT 9 NS 0.0155 0.0059
CxT 9 0.0001 0.0404 0.0001
HxCxT 9 NS NS 0.0355
Ns Nonsignificant at P < 0.05.


Table 3-2. Cultivar by treatment interaction for quantity and length of 2-node stem
cuttings harvested from stock plants of Tecoma stans and Nerium oleander
during the warm season (Expt. 1).
Tecoma Oleander


Length (cm)
6.5
7.4
6.0
5.4
7.1
7.0
6.3
6.4
5.4
4.5


Quantity
13.9
6.8
3.1
1.2
14.5
14.0
14.6
14.7
19.0
17.7


Length (cm)
4.7
5.3
4.9
2.5
3.8
3.7
4.0
4.3
4.0
3.5


Significance **
LSD 2.7
NS, *, *** Nonsignificant or significant at


1.2
P < 0.05, or 0.001,


**
3.3
respectively.


Treatment
Control
Fresco
Fresco
Fresco
FAL-457
FAL-457
FAL-457
Florel
Florel
Florel


Concn
(mg-L1)
0
125
250
500
125
250
500
250
500
1000


Quantity
12.1
9.4
8.8
3.3
10.2
11.1
9.6
7.9
5.8
6.3









Amongst all treatments and between species, the first harvest (9.4) produced 15%

less cuttings than the second harvest (11.0) (LSD=0.2, n=100). For both species,

differences in cutting length were not significant amongst treatments for the first harvest

(5.4 cm). During the second harvest, cuttings harvested from the control had similar

lengths to those harvested from all treatments, except from Fresco at 500 mg-L-1, which

were 57% shorter (data not shown). Stock plants treated with Fresco at 125 or 250 mg-L-
1 produced cuttings that were 5.6 and 4.7 cm, respectively, which were at least 121%

longer than those treated with Fresco at 500 mg-L-1 (2.1 cm). Stock plants treated with

Florel at 250 mg-L-1 (5.0 cm) produced cuttings that were 152% longer than Florel at

1000 mg-L-1 (3.3 cm); both concentrations were similar to Florel at 500 mg-L-1 (3.8 cm)

(LSD=1.64, n=10). Stock plants treated with all concentrations of FAL-457 produced

similar sized cuttings (data not shown).

At least 129% more tecoma cuttings were harvested from the control than from

stock plants treated with Fresco or Florel (Table 3-2). Stock plants treated with FAL-457

produced a similar amount of cuttings to the control. Fresco at 125 mg-L-1 produced at

least 63% fewer cuttings than at the higher concentrations, although cuttings harvested

from Fresco at 250 or 500 mg-L-1 were mostly defoliated with necrotic lesions on stems.

Cutting quantity was similar from all Florel treated stock plants. Cutting length was

similar to the control for all treatments except Florel at 1000 mg-L-1 (Table 3-2). At least

122% longer cuttings were harvested from stock plants treated with Fresco at 125 mg-L-1

than at 250 or 500 mg-L-1. Stock plants treated with Florel at 250 mg-L-1 produced 143%

longer cuttings than at 1000 mg-L-1; however both low and high concentrations were









similar to Florel at 500 mg-L-1. Cuttings harvested from FAL-457 treated stock plants at

any concentration were similar in length.

Differences in tecoma stem diameter were not significant amongst treatments for

the first harvest (0.28 cm). During the second harvest, cuttings from stock plants treated

with Fresco at 125 (0.33 cm) or 250 mg-L-1 (0.35 cm) were at least 138% larger in

diameter than from stock plants treated with Fresco at 500 mg-L-1 (0.24 cm), FAL-457 at

500 mg-L-1 (0.24 cm), or Florel at 250 to 1000 mg-L-1 (0.20 and 0.22 cm) (LSD=0.9,

n=5). All treatments produced similar sized cutting diameters to the control, except stock

plants treated with all concentrations of Florel, which were smaller (data not shown).

Florel treatments at all concentrations were similar to each other, as were FAL-457

treatments.

Stock plants of oleander treated with Florel at 250 mg-L-1 or any concentration of

FAL-457 produced a similar number of cuttings to the control (Table 3-2). Fresco at 125

mg-L-1 produced 219% and 258% more cuttings than at 250 or 500 mg-L-1, respectively,

however cuttings were mostly defoliated and exhibited excessive stem elongation with

signs of necrosis. Florel at 250 mg-L-1 produced 23% fewer cuttings than at 500 mg-L-1,

however both were similar to Florel at 1000 mg-L-1.

Differences in oleander cutting length were not significant amongst treatments (3.8

cm). Cutting stem diameter was similar for all treatments during the first harvest, the

same trend occurred in the second harvest, except for smaller stem diameters with Fresco

(data not shown). Cutting stem diameter was not measured from stock plants treated with

Fresco at 500 mg-L-1, as zero cuttings were produced at the second harvest. Cuttings









harvested from stock plants treated with Fresco at 250 mg-L-1 (0.11 cm) had a 33%

smaller stem diameter than at 125 mg-L-1 (0.16 cm) (LSD=0.05, n=5).

Percent survival and visual rooting quality

The harvest date x species x treatment interaction was significant for percent

survival and rooting quality (Table 3-3). Differences in percent survival were not

significant amongst treatments for the first harvest of tecoma, which exhibited 94%

survival (Table 3-4). Rooting quality of cuttings harvested from the control were similar

to all treatments except Fresco concentrations of 125 and 250 mg-L-1. Fresco at lower

concentrations had reduced rooting quality when compared to all treatments except when

provided at 500 mg-L-1. For the second harvest of tecoma, cuttings planted from stock

plants treated with Fresco at 500 mg-L-1 had at least a 57% lower rate of survival than all

other lateral branching treatments, which performed similarly to the control (Table 3-4).

Rooting quality was similar to the control for cuttings harvested from stock plants treated

with FAL-457 at 125 or 250 mg-L-1 or Florel at 250 or 500 mg-L-1. Fresco at 125 or 250

mg-L-1 had greater root quality ratings than at 500 mg-L-1, which was the lowest of any

treatment. Stock plants treated with Florel at 250 or 500 mg-L-1 had better rooting quality

than 1000 mg-L-1. Cuttings had better rooting quality when planted from stock plants

treated with FAL-457 at 250 mg-L-1, when compared to 125 or 500 mg-L-1.

For the first harvest of oleander, propagules from stock plants treated with Fresco

at 250 or 500 mg-L-1, or FAL-457 at 125, 250, or 500 mg-L-1 exhibited stem and foliar

disease symptoms, and did not survive propagation. While cuttings harvested from stock

plants treated with Florel at all concentrations performed similarly, treatments of 1000

mg-L-1 had a 202% higher survival rate than Fresco treatments at 125 mg-L-1 (Table 3-4).

Rooting quality ratings were higher for cuttings planted from stock plants treated with









Florel at 1000 mg-L-1 than Florel at 500 mg-L1 or Fresco at 125 mg-L1, while Florel at

250 mg-L1 was similar to all three. For the second harvest, no cuttings were planted

from stock plants treated with Fresco at 500 mg-L1. As concentrations of Fresco

increased from 0 to 250 mg-L1, percent survival and subsequent rooting quality

decreased (Table 3-4). Cuttings planted from stock plants treated with Fresco at 250

mg-L1 had the lowest rate of survival, when compared to all other treatments. All

treatments except the two higher concentrations of Fresco had similar survival to the

control. For rooting quality, all treatments were similar to the control, except Fresco at

125 and 250 mg-L1.

Table 3-3. Significance levels for effects of harvest date, cultivar, and lateral branching
treatment on percent survival, visual rooting quality (l=poor; 5=best), total
cutting dry weight (root weight + shoot weight) and root: shoot ratio (root
weight / shoot weight) of cuttings planted from stock plants of oleander and
tecoma during the warm season (Expt. 1).
Percent Rooting Total dry Root:
Source df survival quality weight shoot ratio
Rep 3 NS NS NS NS
Harvest date (H) 1 0.0001 0.0007 NS 0.0001
Cultivar (C) 1 0.0001 0.0001 NS 0.0348
HxC 1 0.0001 0.0001 NS 0.0103
Branching treatment (T) 9 0.0001 0.0001 NS NS
HxT 9 0.0001 0.0001 NS NS
CxT 9 0.0001 0.0001 NS NS
HxCxT 9 0.0124 0.0077 NS NS
Ns Nonsignificant at P < 0.05.

Cutting total dry weight and root: shoot ratio

The harvest date x species interaction was significant for cutting root : shoot ratio

(Table 3-3), however differences in total cutting dry weight were not significant amongst

treatments (0.579 g). For the first harvest, differences in root : shoot ratio between

species were not significant (0.38). For the second harvest however, tecoma (0.29) had a

242% larger root : shoot ratio than oleander (0.12) (LSD=0.07, n=30).












Table 3-4. Harvest date by cultivar by branching treatment interaction for rooting quality (1=poor; 5=best) and percent survival of 2-
node stem cuttings propagated from stock plants of Nerium oleander 'Dwarf Salmon' and Tecoma stans 'Esperanza'
d etsevrah on 3 Aug (H1) and 30 Aug 2005 (H2) during the war )


Tecoma Oleander
Hi H2 Hi H2
Treatment Concn Survival Rooting Survival Rooting Survival Rooting Survival Rooting
(mg-L-1) (%) quality (%) quality (%) quality (%) quality
Control 0 78 4.1 100 4.1 0 0.0 91 3.6
Fresco 125 89 4.2 88 3.4 15 2.0 88 2.8
Fresco 250 100 3.3 96 3.4 0 0.0 13 0.8
Fresco 500 100 3.4 38 2.6 0 0.0 0 0.0
FAL-457 125 100 3.9 100 3.8 0 0.0 96 3.3
FAL-457 250 94 4.1 100 3.2 0 0.0 92 3.4
FAL-457 500 89 4.2 100 4.1 0 0.0 92 2.9
Florel 250 94 4.2 100 4.3 25 3.5 88 3.4
Florel 500 100 4.3 100 4.1 21 2.1 92 3.7
Florel 1000 100 4.1 100 3.3 30 4.4 95 3.4

Significance NS *** *** *** *** *
LSD 0.6 15 0.5 15 1.8 19 0.8
Ns, *, ** Nonsignificant or significant at P < 0.05 or 0.001, respectively.









Experiment 2

Harvest yield, cutting length, and stem diameter

There was a harvest date x species x treatment interaction for all measured

parameters except stem diameter (Table 3-5). The harvest x species and harvest x

treatment interactions were both significant for stem diameter.

Table 3-5. Significance levels for effects of harvest date, cultivar, and branching
treatment on quantity, length, and stem diameter of cuttings harvested from
stock plants of oleander and tecoma during the cool season (Expt. 1).
Source df Quantity Length Diameter
Rep 8 0.0001 0.0316 NS
Harvest date (H) 1 0.0001 NS NS
Cultivar (C) 1 0.0001 0.0198 NS
HxC 1 0.0001 0.0001 0.0021
Branching treatment (T) 9 0.0003 0.0001 0.0095
HxT 9 0.0417 NS 0.0108
CxT 9 0.0001 0.0001 NS
HxCxT 9 0.0001 0.0020 NS
Ns Nonsignificant at P < 0.05.


For the first harvest of tecoma, stock plants treated with FAL-457 at 125 to 250

mg-L-1 or Fresco at 125 mg-L1 produced a similar number of cuttings to the control

(Table 3-6). Untreated plants produced at least 143% more cuttings than stock plants

treated with Fresco at 250 or 500 mg-L1, FAL-457 at 500 mg-L1, or Florel at any

concentration. Florel at 250 mg-L1 produced 520% more cuttings than at 1000 mg-L1,

but was similar to 500 mg-L1. Similarly, Fresco at 125 mg-L1 produced 178% more

cuttings than 500 mg-L1, yet quantity was similar to 250 mg-L1.

Tecoma stock plants treated with Florel at 1000 mg-L1 produced the smallest

number of cuttings (Table 3-6). All other treatments performed similarly to the control,

except Fresco at 125 and 500 mg-L1, which yielded at least 131% more cuttings. For the

second harvest, cutting quantity was similar to the control for all treatments except Florel









at 1000 mg-L-1 (Table 3-7). Within chemical, cutting yield was similar, however upon

comparison of lateral branching agents at least a 181% greater number of cuttings were

harvested from stock plants treated with FAL-457 at 125 or 250 mg-L-1 than from those

treated with all concentrations of Fresco or Florel at 1000 mg-L-1. Cutting quantity

harvested from stock plants treated with all concentrations of Florel was similar to all

concentrations of Fresco.

Table 3-6. Influence of lateral branching agents on number and length of cuttings and
subsequent rooting performance values for cuttings harvested on from stock
plants of Tecoma stans 'Esperanza' on 28 Nov. 2005.
Concn Length Rooting Total dry Root: shoot
Treatment (mg-L-1) y (cm) quality weight (g) ratio
Control 0 7.0 4.9 4.0 0.488 0.24
Fresco 125 6.3 7.7 3.8 0.460 0.38
Fresco 250 4.9 6.3 3.1 0.296 0.29
Fresco 500 3.6 6.5 2.8 0.219 0.25
FAL-457 125 7.0 5.3 3.5 0.478 0.24
FAL-457 250 5.9 5.3 2.8 0.367 0.22
FAL-457 500 3.8 5.3 3.2 0.386 0.18
Florel 250 2.9 5.5 4.0 0.338 0.30
Florel 500 1.8 4.3 2.8 0.181 0.35
Florel 1000 0.6 1.8 4.2 0.244 0.34

Significance ** ** *
LSD 1.9 1.5 0.9 0.142 0.13
Significant at P < 0.05 or 0.01.

During the first harvest, cutting length was similar for all treatments as compared to

the control, except those from stock plants treated with Fresco at 125 or 500 mg-L-1,

which had longer cuttings (Table 3-6). Fresco at 125 mg-L-1 had longer cuttings than any

other treatment, except at concentrations of 250 to 500 mg-L-1. When comparing

individual branching agents, cutting length was similar for all concentrations of Fresco,

as were all concentrations of FAL-457. Stock plants treated with concentrations of Florel

at 250 to 500 mg-L-1 produced at least 241% longer cuttings than at 1000 mg-L-1. Cutting









length from the second harvest was similar to the control for all treatments except Fresco

at 500 mg-L-1 (Table 3-7). Cuttings harvested from stock plants treated with Fresco at

500 mg-L-1 were 372% longer than from Florel at 1000 mg-L-1. When considering each

branching agent individually, cutting length was similar for all concentrations of each

chemical applied.

Table 3-7. Influence of lateral branching agents on quantity and length of cuttings and
subsequent dry weights of cuttings harvested on from stock plants of Tecoma
stans 'Esperanza' on 28 Dec. 2005.
Concn Length Total dry Root:
Treatment (mg-L-1) Quantity (cm) weight (g) shoot ratio
Control 0 3.4 2.5 0.110 0.55
Fresco 125 2.3 4.0 0.210 0.30
Fresco 250 1.8 3.0 0.113 0.39
Fresco 500 2.1 4.9 0.140 0.28
FAL-457 125 4.2 3.4 0.271 0.15
FAL-457 250 4.2 3.7 0.266 0.15
FAL-457 500 3.3 3.5 0.211 0.26
Florel 250 2.6 2.6 0.055 0.50
Florel 500 2.6 2.4 0.037 0.99
Florel 1000 1.2 1.3 0.015 1.64

Significance ** *
LSD 1.8 1.9 0.079 0.71
** Significant at P < 0.05 or 0.001.

At the first harvest of oleander, cutting quantity was similar for stock plants treated

with Fresco at 125 mg-L-1, Florel at 1000 mg-L-1, or FAL-457 at all concentrations (Table

3-8). Additionally, stock plants treated with Fresco at 125 to 250 mg-L-1 or Florel at 250

to 500 mg-L-1 generated at least 308% more cuttings than untreated stock plants. Results

varied in response to concentration for Florel and Fresco, while FAL-457 at all

concentrations performed similarly. Florel at 250 to 500 mg-L-1 produced more cuttings

than at 1000 mg-L-1, while Fresco at 125 mg-L-1 produced fewer cuttings than at









concentrations of 250 to 500 mg-L-1. During the second harvest, with 9.4 cuttings per

plant, differences in quantity were not significant amongst treatments.

Oleander cutting length during the first harvest was similar for all branching agents

as compared to the control, except Fresco at 250 and 500 mg-L-1 (Table 3-8). Fresco

treatments of 125 mg-L-1 produced 38% shorter cuttings than 500 mg-L-1, while both

concentrations were similar to 250 mg-L-1. Cutting length in response to all

concentrations of Florel and FAL-457 were similar. During the second harvest, lengths

of cuttings from untreated plants were similar to FAL-457 at all concentrations and Florel

at 500 mg-L-1 (Table 3-9). All concentrations of Florel produced shorter cuttings than

cuttings from Fresco treated stock plants. Longer cuttings with Fresco at 250 mg-L-1

were recorded when compared to 500 mg-L-1.

When considering both species, at the first harvest and amongst treatments, cutting

diameter was 126% larger for tecoma (0.23 cm) than oleander (0.19 cm) (LSD=0.04,

n=90). At the second harvest however, oleander (0.20 cm) had a 118% larger diameter

than tecoma (0.17 cm) (LSD=0.02, n=90). During the first harvest of both species, all

branching agents provided a similar diameter to the control (0.18 cm), except Florel at

250 mg-L-1 (0.33 cm) which was at least 145% larger than any other treatment (LSD=0.1,

n=18). Stock plants treated with Fresco at 500 mg-L-1 (0.23 cm) produced a 183% larger

diameter than those treated with Florel at 1000 mg-L-1 (0.12 cm), although both were

similar to the control. At the second harvest, most treatments were similar to the control

(0.21 cm) except Fresco at 125 mg-L-1 (0.16 cm) or 250 mg-L-1 (0.15 cm) or Florel at 250

or 1000 mg-L-1 (0.17 and 0.13 cm, respectively) (LSD=0.05, n=18). Stock plants treated

with FAL-457 at 125 mg-L-1 (0.24 cm) or 500 mg-L-1 (0.22 cm) produced cuttings with at









least 125% greater diameter than from those treated with Fresco or Florel at any

concentration (data not shown). Relative differences amongst concentrations for each

chemical were not significant.

Percent survival and visual rooting quality

There was a harvest date x cultivar x treatment interaction for both percent survival

and visual rooting quality (Table 3-10). Differences in percent survival were not

significant amongst treatments for cuttings planted from either harvest of tecoma, which

were 100% and 77% from harvests 1 and 2, respectively. Rooting quality from the first

harvest of untreated cuttings was similar to most treatments except concentrations of

Fresco at 250 to 500 mg-L-1, FAL-457 at 250 mg-L-1, and Florel at 500 mg-L-1, which had

lower quality ratings (Table 3-6). Florel treatments at 250 and 1000 mg-L-1, were similar,

yet produced higher quality ratings than at 500 mg-L-1. Although Fresco at 125 mg-L-1

had lower rooting quality than at 500 mg-L-1, both were similar in quality to Fresco at 250

mg-L-1. All concentrations of FAL-457 performed similarly to each other. Florel at 1000

mg-L-1 had higher root quality ratings than cuttings from stock plants treated with Fresco

or FAL-457 at 250 or 500 mg-L-1, respectively. Differences in rooting quality for tecoma

cuttings from the second harvest were not significant amongst treatments (2.2).

Branching agents did not improve percent survival for oleander cuttings planted

from the first harvest as compared to the control (Table 3-8). Cuttings planted from stock

plants treated with FAL-457 at 250 mg-L-1, or Florel at 250 to 500 mg-L-1, survived better

than those treated with Fresco at 500 mg-L-1, FAL-457 at 125 mg-L-1, or Florel at 1000

mg-L-1.

Cuttings had the highest rooting quality values from untreated stock plants than

from all lateral branching agents applied (Table 3-8). FAL-457 treatments provided









similar rooting quality at all concentrations; however, FAL-457 cuttings had lower

rooting quality than Florel treatments at 500 or 1000 mg-L-1. Fresco had similar rooting

quality values to Florel.

Table 3-8. Influence of lateral branching agents on quantity and length, and subsequent
rooting for cuttings harvested from stock plants of Nerium oleander 'Dwarf
Salmon' on 28 Nov. 2005.
Root :
Concn Quantity Length Survival Rooting Total dry soot:
Treatment (m -1 (cm) (%) quality weight (g) ratio
Treatment (mg-L ) ratio
Control 0 1.3 2.4 93 4.4 0.522 0.13
Fresco 125 1.9 4.1 93 2.8 0.277 0.27
Fresco 250 4.8 5.4 91 2.8 0.202 0.17
Fresco 500 4.1 6.6 73 2.6 0.112 0.22
FAL-457 125 2.4 2.6 73 2.4 0.317 0.15
FAL-457 250 1.7 1.8 100 1.8 0.293 0.06
FAL-457 500 2.0 2.1 83 1.8 0.281 0.07
Florel 250 5.6 2.1 96 2.6 0.170 0.17
Florel 500 4.2 2.0 100 3.1 0.151 0.22
Florel 1000 1.6 1.6 72 3.2 0.065 0.38

Significance ** *
LSD 2.0 1.8 21 0.6 0.134 0.11
*, *** Significant at P < 0.05 or 0.001.


For the second harvest of oleander, cuttings generated from the control had similar

percent survival to all branching treatments except Fresco concentrations of 125 to 500

mg-L-1 (Table 3-9). Cuttings planted from stock plants treated with increasing

concentrations of Fresco experienced drastic decreases in percent survival. Fresco at 500

mg-L-1 had the lowest percent survival. Rooting quality for cuttings propagated from

stock plants treated with FAL-457 at 125 mg-L-1 was similar the control, while all other

branching treatments were lower in quality (Table 3-9). When considering each chemical

individually, Fresco treated cuttings declined in rooting quality as the concentration

increased, while concentrations of FAL-457 at 250 to 500 mg-L-1 performed similarly,









but with poorer quality as compared to 125 mg-L-1. Rooting response to Florel at 250

mg-L-1 was similar to 500 mg-L-1. Cuttings harvested from stock plants treated with

Florel at 500 mg-L-1 had higher rooting quality than those treated with Florel at 1000

mg-L-1 or Fresco at 250 mg-L-1. Cuttings planted from stock plants treated with Fresco at

500 mg-L-1 had the poorest rooting quality when compared to all other treatments.

Table 3-9. Influence of lateral branching agents on number and length of cuttings, and
subsequent rooting performance values for cuttings harvested on from stock
plants of Nerium oleander 'Dwarf Salmon' on 28 Dec. 2005.
Concn Length Survival Rooting Total dry Root: shoot
Treatment (mg-L-1) (cm) (%) quality weight (g) ratio
Control 3.9 100 4.4 0.539 0.20
Fresco 125 6.7 83 3.0 0.187 0.17
Fresco 250 8.1 37 1.9 0.091 0.17
Fresco 500 6.2 9 1.0 0.108 0.43
FAL-457 125 4.7 100 4.2 0.439 0.20
FAL-457 250 4.2 100 2.8 0.561 0.11
FAL-457 500 3.9 100 3.0 0.590 0.09
Florel 250 3.9 100 3.2 0.217 0.28
Florel 500 2.6 100 3.3 0.181 0.24
Florel 1000 1.9 100 2.5 0.074 0.42

Significance **
LSD 1.5 11 0.8 0.092 0.09
Significant at P < 0.05 or 0.001.


Cutting total dry weight and root : shoot ratio

There was a harvest date x cultivar x treatment interaction for total cutting dry

weight and root : shoot ratio (Table 3-10). Tecoma cuttings planted from the first harvest

were similar in total cutting dry weight to the control when stock plants were provided

Fresco at 125 mg-L-1 or any concentration of FAL-457 (Table 3-6). Cuttings planted

from stock plants treated with Fresco at concentrations of 250 to 500 mg-L-1, were at least

36% lower in total dry weight than at 125 mg-L-1. Dry weights for all concentrations of

FAL-457 were similar, as were all concentrations of Florel. Overall, Fresco or FAL-457









at low concentrations had larger dry weights than Florel at 500 to 1000 mg-L-1. Root :

shoot ratios for all branching treatments were similar to the control, except Fresco at 125

mg-L-1, which was 161% greater (Table 3-9). The root: shoot ratio of Fresco at 125

mg-L-1 was greater than at 500 mg-L-1 as well as all concentrations of FAL-457. Fresco

and Florel were similar with respect to the cutting root : shoot ratio, except when FAL-

457 was applied at 125 mg-L-1.

For the second harvest, total cutting dry weight of tecoma cuttings propagated from

stock plants treated with Fresco at 250 mg-L-1 or Florel at 250 to 500 mg-L-1 were similar

to the control (Table 3-7). When considering FAL-457, all concentrations had similar

dry weights, yet when stock plants were provided concentrations of 125 to 250 mg-L-1,

total cutting dry weights were at least 194% greater than when planted from Fresco

treated stock plants at 250 to 500 mg-L-1 or all concentrations of Florel. FAL-457 also

had increased cutting dry weight as compared to the control. Cuttings showed a

reduction in total cutting dry weight when treated with Fresco at 250 to 500 mg-L-1 as

compared to 125 mg-L-1. Fresco at 250 mg-L-1 was similar to Florel at 500 to 1000 mg-L-

1

Branching agents applied to tecoma stock plants at all concentrations produced

similar root: shoot ratios as compared to the control, except Florel at 1000 mg-L-1 (Table

3-7). Root: shoot ratios of Florel at 500 to 1000 mg-L-1 were at least 355% higher than

Fresco at 500 mg-L-1 or any concentration of FAL-457; however, higher concentrations

of Florel caused stunting with little to no shoot growth during propagation. Florel at 250

mg-L-1 performed similarly to 500 mg-L-1 and any concentration of Fresco.









Oleander cuttings from the first harvest had at least 165% greater total dry weight

with the untreated control than from any other lateral branching agent (Table 3-8). When

considering each chemical individually, all concentrations of Florel had similar dry

weights, as did FAL-457. Fresco had greater dry weight at 125 mg-L-1 than 500 mg-L-1,

yet both concentrations were similar to 250 mg-L-1. Cuttings planted from stock plants

treated with FAL-457 at 250 or 500 mg-L-1 had at least 173% greater dry weight than

plants treated with Fresco at 500 mg-L-1 or Florel at any concentration.

Oleander cutting root : shoot ratios were similar to the control for all treatments

except for Fresco at 125 mg-L-1 or Florel at 1000 mg-L-1 which had larger ratios (Table 3-

8). When comparing individual chemicals, all concentrations of Fresco had a similar root

: shoot ratio, as did all concentrations of FAL-457. Florel at 250 or 500 mg-L-1 had a

lower root: shoot ratio than 1000 mg-L-1. There was at least a 166% higher root: shoot

ratio for cuttings propagated from plants treated with Florel at 1000 mg-L-1 than from

untreated plants, all concentrations of FAL-457, or Fresco and Florel at 250 to 500 mg-L-

1, respectively.

For the second harvest of oleander, cuttings propagated from stock plants treated

with FAL-457 at 250 or 500 mg-L-1 produced cuttings with a similar total dry weight to

the control (Table 3-9). Cuttings planted from untreated stock plants and from those

treated with concentrations of FAL-457 at 250 or 500 mg-L-1 were at least 123% greater

in total cutting dry weight than plants treated with FAL-457 at 125 mg-L-1. FAL-457 at

250 or 500 mg-L-1 were also 248% greater than any concentration of Fresco or Florel.

Florel had reduced total cutting dry weight at 1000 mg-L-1 as compared to either 250 or

500 mg-L-1. Cuttings rooted from Fresco treated stock plants had greater dry weight at









125 mg-L-1 than at 250 mg-L-1, while both of these concentrations were similar to 500

mg-L-1.

All treatments had a similar root : shoot ratio as compared to the control, except

FAL-457 at 250 mg-L-1, which was smaller, and Fresco and Florel at 500 and 1000 mg-L-

, respectively, which were both larger. The cutting root: shoot ratio was at least 149%

greater for cuttings propagated from stock plants treated with Fresco at 500 mg-L-1 or

Florel at 1000 mg-L-1 than from the all other treatments (Table 3-9). Stock plants treated

with Florel at 250 mg-L-1 had rooted cuttings with at least 162% greater root : shoot ratio

than from plants treated with Fresco at 125 to 250 mg-L-1, or FAL-457 at 250 to 500

mg-L-1. When comparing individual branching agents, Fresco at 125 to 250 mg-L-1 had a

lower root : shoot ratio than 500 mg-L-1. Results were similar with Florel concentrations;

however, FAL-457 at 125 mg-L-1 was greater than 500 mg-L-1, while both produced

similar results to 250 mg-L-1.

Discussion

During the first experiment, when data was pooled between species to investigate

the harvest by treatment interaction, both tecoma and oleander had larger yield responses

at the second harvest as compared to the first, which suggests increased cutting

production in response to pinching, rather than chemically influenced. Regardless of

treatment, oleander stock plants generated more cuttings than tecoma. Lateral branching

agents tended to not improve cutting yield of tecoma, rather decreases in yield occurred

with Florel and Fresco applications. Oleander also negatively responded to Fresco at all

concentrations, however yield was enhanced when stock plants were treated with Florel.

All concentrations of FAL-457 produced similar numbers of cuttings to untreated plants

for both species. The application of a lateral branching agent produced similar sized









oleander cuttings, when compared to the control, however slightly shorter cuttings than

the control were produced when Florel was provided at increasing concentrations to

tecoma. Stem diameter was generally not influenced by application of a branching agent,

although Florel treatments had smaller stem diameters than untreated cuttings during the

second harvest of both species.

Application of branching agents did not influence percent survival of tecoma from

the first harvest, while the second harvest produced cuttings with a lower survival rate

when Fresco was provided to stock plants as compared to the control. Subsequent

rooting quality was also reduced for cuttings planted from Fresco treated stock plants,

while all other treatments were similar to the control when planted from both harvests.

Phytotoxicity may have increased disease incidence, however similar symptoms

were observed with the control, which may be result of disease pressure. From the

second harvest phytotoxicity of Fresco sprays were evident as percent survival and

subsequent rooting quality decreased as concentrations of Fresco increased, otherwise

rooting was similar for all treatments, when compared to the control. Dry weight was

generally similar for all treatments and for both species, although tecoma cuttings from

the second harvest had a larger root : shoot ratio than oleander.

During the second study, phytotoxicity in response to treatment applications was

similar to the first experiment, although overall performance of cuttings during the

second experiment was generally improved by comparison.

During the first harvest, tecoma showed a reduction in cutting quantity with

increasing concentrations of branching agents applied, although Fresco and FAL-457 at

lower rates tended to be similar to the control. Length of cuttings was excessive when









stock plants were treated with Fresco and stunted when treated with Florel, as compared

to cuttings harvested from untreated stock plants. With few exceptions, branching agents

applied at higher concentrations to stock plants of tecoma caused a reduction in

subsequent rooting quality and total cutting dry weight when compared to the control.

Root : shoot ratios however, tended to be similar to the control for all treatments,

improved ratios occurred when Fresco or Florel was applied at 125 or 500 mg-L-1,

respectively.

A reduction in cutting number occurred for all treatments during the second

harvest, when compared to the first. The number of tecoma cuttings was similar for both

treated and untreated stock plants, except when treated with Florel at 1000 mg-L-1.

Cutting length was also shorter during the second harvest, when compared to the first.

Supplemental applications of lateral branching agents produced similar length cuttings

except with Fresco at 500 mg-L-1. Stock plants treated with Fresco at 500 mg-L-1

produced cuttings with the same length (4.9 cm) as the untreated stock plants during the

first harvest; therefore no cuttings would be considered excessive in length from the

second harvest.

Benefits to supplemental application of branching agents to stock plants were not

shown with percent survival and rooting quality because all treatments were similar to the

untreated cuttings. Differences in total cutting dry weight follow a similar trend with

differences in cutting length. Cuttings from stock plants treated with Florel at 1000

mg-L-1 were shorter than the control, but also lower total cutting dry weight. Phytotoxic

responses such as shoot tip desiccation, defoliation, or excessive stunting as a result of

Florel applied at 1000 mg-L-1 caused an impractically high root : shoot ratio, otherwise all









branching treatments were similar to the control. In studies conducted on other

floriculture species, undesirable stunting has been reported in response to Florel (Faust

and Lewis, 2005).

During the first harvest of oleander, cutting quantity was improved when Fresco

and Florel were applied at 250 to 500 mg-L-1; all other treatments were similar to the

control. Compared to tecoma, oleander stock plants did not show the same reduction in

yield with application of branching agents. Increasing concentrations of Florel applied to

stock plants decreased yield, although quantities harvested were still greater than or equal

to the control. Generally lengths of cuttings were similar to the control, yet were

excessive when harvested from stock plants treated with Fresco at higher concentrations.

All treatments rooted with similar percent survival when compared to the control,

except for reduced survivability with Florel at 1000 mg-L-1. Despite similar percent

survival, all treatments had a reduced visual rooting quality when compared to the

control. Among the least affected by treatment with respect to root quality rankings were

those harvested from stock plants treated with Fresco and lower concentrations of FAL-

457 or Florel. Both total cutting dry weight and the root: shoot ratio seem to coincide

with differences observed with rooting quality.

During the second harvest, all treatments produced the same number of cuttings,

yet comparable to other harvests, cutting length was similar for most treatments with

stunting and stretching of propagules when stock plants were treated with Florel or

Fresco, respectively. Percent survival was 100% for all treatments except poor survival

rates associated with increasing concentrations of Fresco. Rooting quality was also

reduced with all Fresco concentrations and quality declined as concentration increased.









Total cutting dry weight was lower than the control for all treatments except FAL-457 at

250 to 500 mg-L-1, which were similar. Lower total cutting dry weights, coupled with

increased root: shoot ratios for both Fresco at 500 mg-L-1 and Florel at 1000 mg-L-1

reflected desiccation of the shoot tip or stunting, respectively. Similar root : shoot ratios

for nearly all other treatments were similar, which may be a better indication of rooting

performance relative to cutting size than visual evaluations, therefore oleander cuttings

performed similarly to the control when treated with branching agents prior to the second

harvest.

Fresco applications to the stock plants exhibited many phytotoxic symptoms that

would be considered highly undesirable for cutting propagation. Similar phytotoxic

responses were observed when Fresco was applied to stock plants of Clematis spp. and

Rhododendron spp. (Bell et al., 1997; Puglisi, 2002). Symptoms were more severe on

oleander than tecoma. The undesirable effects of BA+GA4+7 (active ingredient in Fresco)

have been attributed to spray burn as a result of BA in the treatment formulation (Bell et

al., 1997). When Clematis spp., were treated with different ratios of BA and GA to

reduce phytotoxicity, all combinations had similar effects. The 1:1 ratio (similar

formulation to Fresco) produced the lowest degree of phytotoxicity (Puglisi, 2002).

Cuttings harvested from stock plants treated with FAL-457 did not express the same

symptoms; therefore it is hypothesized that the phytotoxicity is a species-dependant

outcome, and may be the result of other components such as the chemical carrier.

Concentrations of FAL-457 at 125 mg-L-1 were shown to improve yield in

oleander, but reduced success in propagation limits its use by growers. In tecoma, FAL-

457 produced similar or lower quality cuttings than the untreated plants. Although









application of FAL-457 was not shown to be consistently beneficial to either species;

further investigation should be conducted to determine the efficacy of FAL-457 as a

possible tank mix with other formulations. Perhaps FAL-457 and Florel could be mixed

to capitalize on the benefits to yield and rooting of both treatments.

Applications of Florel at 250 to 500 mg-L-1 to stock plants of oleander were shown

to increase yield with similar rooting when compared to the control, yet results were not

consistent. Florel applications to stock plants of tecoma were not shown to be

advantageous. Variability in response to Florel for increasing cutting quantity has been

reported with many floriculture species (Faust and Lewis, 2005), therefore benefits to its

application may been seen at lower concentrations or with other tropical species. Further

study should be conducted on the efficacy of these and other lateral branching agents

during other seasons and on other tropical species.

Conclusion

At the concentrations investigated, none of the lateral branching agents was proven

to be consistently advantageous when compared to untreated plants. Phytotoxic

responses were shown that could potentially damage crops in production, therefore

growers should consider testing sprays on a smaller portion of plants before applications

are made to the entire greenhouse. The cost of applying these products should be

compared to the benefits achieved in production. While tecoma and oleander appeared to

not benefit from branching treatments at the concentrations investigated, trends were

observed that may assist future research.









Table 3-10. Significance levels for effects of harvest date, cultivar, and lateral branching
treatment on percent survival, visual rooting quality (l=poor; 5=best), total
cutting dry weight (root weight + shoot weight) and root: shoot ratio (root
weight / shoot weight) of cuttings planted from stock plants of oleander and
tecoma during the cool season (Expt. 2).
Percent Rooting Total dry Root:
Source df survival quality weight shoot ratio
Rep 3 NS NS 0.0106 NS
Harvest date (H) 1 0.0001 0.0001 0.0001 0.0004
Cultivar (C) 1 NS NS NS 0.0001
HxC 1 0.0012 0.0001 0.0001 0.0198
Branching treatment (T) 9 0.0001 0.0001 0.0001 0.0001
HxT 9 0.0001 0.0004 0.0001 0.0257
CxT 9 0.0001 0.0001 0.0001 0.0454
HxCxT 9 0.0009 0.0001 0.0013 0.0278
NS Nonsignificant at P < 0.05.














CHAPTER 4
ROOTING HORMONES

Introduction

Vegetative propagation has become an established system for reproducing tropical

plants. Treating stem cuttings with auxin increases rooting percentage, accelerates root

formation, and enhances uniformity of rooting (Davis et al., 1988). This is important in

the production of tropical plants because many species often require extensive rooting

time in propagation where there is an inherent risk of air and waterborne pathogens

(Howard, 1994). In order to limit time in propagation, studies have been conducted on

the stimulatory influence of auxin on numerous difficult-to-root species (Bhattacharjee

and Balakrishna, 1983; Bhat et al., 1988; Czekalski, 1989). Preference for the sythentic

compounds indolebutyric acid (IBA) and napthaleneacetic acid (NAA) compared to

naturally occurring, indoleacetic acid (IAA), is illustrated by the large number of

commercially-available rooting products containing one or both in solution (Blazich,

1988a). There are also potassium salt (K+) formulations available that enable IBA

(KIBA) and NAA (KNAA) to be dissolved in water, which may be beneficial for some

tropical species expressing sensitivity to alcohol-based formulations. Certain tropical

species, such as oleander (Nerium oleander L.) and bougainvillea (Bougainvillea glabra

Choisy.), have been shown to benefit from applications of talc-based formulations at

concentrations of 3,000 mgL-1 IBA and 4,000 to 16,000 mgL-1 IBA, respectively

(Hartmann et al., 2002). For other difficult-to-root tropical species an evaluation of









commercially-available rooting hormones should be conducted to determine the optimum

concentration to achieve the highest rooting percentage.

The objectives of our experiment were to determine the optimum concentration of

water-soluble KIBA necessary to effectively root stem cuttings of select tropical species

and to compare the viability of KIBA as an alternative rooting hormone to an industry

formulation containing IBA 1%: NAA 0.5% at 1500 mg-L-1.

Materials and Methods

Experiment 1

Rooted liners of Allamanda schottii (Pohl.), Bougainvillea glabra (Choisy.)

'California Gold' and 'Helen Johnson', Mandevilla splendens (Hook. f.) 'White', and

Nerium oleander (L.) 'Dwarf Salmon' were planted, 3-per pot, into 16 d x 18.5 h cm

round plastic containers on 18 April 2005. The root substrate used was Fafard 4-P

(Fafard, Anderson, S.C.), which contained: 4 sphagnum peat: 2 pine bark: 2 vermiculite:

1 perlite (v/v). Controlled release fertilizer (19N-2.6P-9.9K) was incorporated into the

substrate prior to planting, at a rate of 3.21 g-m-3 (1.33 oz/ft3). The plants were then

provided a once-weekly liquid fertilization of 14N-1.7P-11.6K with N at 200 mg-L-1.

Plants were established for 4 weeks, until roots reached the bottom of the pot, and then

provided a soft pinch to encourage canopy growth on 16 May. Cuttings were harvested

every three weeks, on 6 June (HI), 27 June (H2), 18 July (H3), and 8 Aug. (H4).

Mandevilla was harvested three times on different dates with HI, H2, and H3 occurring 6

June, 8 Aug. and 29 Aug., respectively. Photosynthetic photon flux (PPF) was recorded

each day at or near solar noon using a Quantum Meter (Spectrum Technologies, East-

Plainfield, Ill.). Average PPF was 453, 586, and 426 tmol.m-2S-1, for June, July, and

Aug., respectively.









Harvesting procedure

At each harvest, 6.0 cm terminal stem cuttings were removed from stock plants of

allamanda and oleander. Sub-terminal bougainvillea 'California Gold' and 'Helen

Johnson' stem cuttings were harvested in lengths of 6.5 cm, measured 0.5 cm above first

mature leaf. Mandevilla was harvested by removing all 2-node, sub-terminal, stem

cuttings 1 cm above and below the top and bottom nodes. Cuttings, with bottom leaves

removed, were inserted into perforated plastic bags with stem bases wrapped in moist

paper towels. The bags were placed into boxes, along with a sensor (Hobo Pro, model

number H08-032-08) (Onset Computer, Bourne, Mass.) that recorded temperature and relative

humidity every 15 s. The boxes were stored overnight in a cooler at 10 C. Boxes were

sealed and then cuttings were shipped 24 h, to simulate industry handling. Upon arrival,

boxed cuttings were left intact until the following morning, unpacked and treated with

rooting hormones before planting, totaling approximately 72 h from excision to planting

(Table 4-1). Cuttings were dipped 0.5 cm, for 3s, in a solution containing 1%

indolebutyric acid (IBA) : 0.5% napthaleneacetic acid (NAA) (Dip n' Grow, Dip n' Grow

Inc., Clackamas, Ore.) at 1,500 mgL-1 or Indolebutyric acid, potassium salt (KIBA) at

1,500, 3,000, or 6,000 mg-L-1 and then inserted in a substrate containing 4 perlite: 1

vermiculite (v/v). Cuttings were assigned to a completely randomized design (CRD) and

propagated under mist (5s every 20 min). Each species was randomized separately and

was assigned, when numbers were available, with 6 sub-samples and 4 replications of

each treatment. Sub-sample means were pooled to reduce variability of the replicate.

Cutting evaluations

Cuttings were evaluated on two occasions from each harvest. Allamanda was

evaluated 2 and 4 weeks after planting (WAP), oleander and mandevilla 4 and 6 WAP,









and both bougainvillea cultivars 5 and 7 WAP. At the first evaluation of each harvest

percent survival, rooting quality, number of primary roots (>1.0 mm), number of lateral

roots, total cutting dry weight (root weight + shoot weight), and cutting root : shoot ratio

(root weight/ shoot weight) were recorded. At the second evaluation all other parameters

except root number were recorded. Visual rooting quality ratings were assigned on a 1 to

5 scale, relative to each species, with a general ranking system as follows: l=callus

formation, with no roots; 2=minimal rooting; 3=some root growth and development;

4=increased root growth and development; 5= best rooting performance. Cuttings were

prepared for dry weight measurement by excision of the rooted portion 1 cm above the

cutting base, and then placed in a drying oven for 3 d at 70 C. All data were subjected to

analysis of variance using generalized linear model procedure in SAS (SAS institute,

Cary, N.C.) and means separated using LSD with P < 0.05.

Experiment 2

The second experiment was replicated in the same manner as the first experiment,

with the following exceptions: Stock plants were transplanted 1 Aug., and given a soft

pinch on 17 Aug. 2005. Cuttings were again harvested every three weeks on 6 Sept.

(HI), 27 Sept. (H2), 18 Oct. (H3), and 8 Nov. (H4). Average PPF was 596, 647, and 355

tmol.m-2S-1, for Sept., Oct., and Nov., respectively. Cuttings were handled in the same

manner as the first experiment, except were shipped to Gainesville, Fla. using Federal

Express 24 h service (Table 4-1).









Results

Allamanda schottii

Experiment 1

There was a harvest date x evaluation week x treatment interaction for percent

survival (Table 4-2). Differences in percent survival were not significant amongst

treatments for all cutting evaluations, except those from H3, 2 WAP and H4, 4 WAP.

For cuttings planted from H3 and evaluated 2 WAP, all rooting hormone concentrations

provided similar percent survival and were greater than the control, except those treated

with KIBA at 1500 mg-L-1, which were similar to all treatments. Cuttings evaluated 4

WAP from H4 were all similar to the control when treated with any rooting hormone

concentration except Dip n' Grow at 1500 mg-L-1, which had better survival.

Concentrations of KIBA at 3000 mg-L-1 had similar percent survival to Dip n' Grow at

1500 mg-L-1, and expressed greater survivability than KIBA at 1500 or 6000 mg-L-1.

Generally, cuttings evaluated 4 WAP had a similar percent survival to 2 WAP, except

those treated with KIBA at 6000 mg-L-1 and planted from H4, which were greater 2 WAP

than 4 WAP. When evaluating cuttings 2 WAP, all treatments had a similar response

throughout the experiment; some differences occurred with respect to harvest date 4

WAP. For untreated cuttings or cuttings treated with KIBA at 3000 mg-L-1, all harvest

dates had similar percent survival 4 WAP. Cuttings treated with Dip n' Grow at 1500

mg-L-1 had similar percent survival at all harvest dates except H3, which was lower.

When cuttings treated with either KIBA at 1500 or 6000 mg-L-1 were evaluated 4 WAP,

all harvest dates were similar, except H4, which produced lower survival rates.

There was a harvest x treatment interaction for rooting quality (Table 4-3); main

effects for evaluation week will be presented separately. Cuttings evaluated 4 WAP had









higher root quality rankings than at 2 WAP (data not shown). Differences in rooting

quality were not significant amongst treatments for HI, H2, or H4. For cuttings planted

from H3, rooting hormone applications at any concentration had better rooting quality,

when compared to the control, except those treated with KIBA at 1500 mg-L-1. Rooting

quality of cuttings treated with KIBA at 1500 mg-L-1 was similar to all treatments except

was lower than KIBA at 3000 mg-L-1. Cuttings treated with KIBA at 3000 mg-L-1 had

greater rooting quality than cuttings treated with KIBA at 1500 mg-L-1 or the control.

When considering cuttings that were untreated or treated with KIBA at 1500 mg-L-1,

those planted from H4 were similar to all other harvests, while cuttings planted from HI

or H2 had greater rooting quality than H3. When cuttings were treated with Dip n' Grow

at 1500 mg-L-1, all harvest dates were similar, except H3. Rooting quality was similar for

all harvest dates when treated with KIBA at 3000 or 6000 mg-L-1, respectively.

Table 4-3. Harvest date (H) by treatment interaction for visual rooting quality (1-5;
l=poor, 5=best) of allamanda cuttings planted during the warm season (Expt.
1).
Concn (mg-L-
Treatment 1) HI H2 H3 H4 LSD
Control 0 3.9 3.5 2.5 3.2 0.9
Dip n' Grow 1500 4.1 4.3 3.2 4.2 ** 0.5
KIBA 1500 4.2 3.9 3.1 3.7 0.7
KIBA 3000 4.1 4.1 3.9 3.9 NS
KIBA 6000 4.3 4.3 3.8 3.8 NS

Significance NS NS ** NS
LSD 0.7
NS, *, ** Nonsignificant or significant at P < 0.05, 0.01, 0.001, respectively.
Dip n' Grow = 1% indolebutyric acid : 0.5% napthaleneacetic acid
KIBA = indolebutyric acid, potassium salt


There was a harvest date x treatment interaction for number of primary roots. From

H1, cuttings treated with KIBA at 1500 to 3000 mg-L-1 had a similar number of primary