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Integrated Control of Tropical Signalgrass (Urochloa Subquadripara) in Turf

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Title:
Integrated Control of Tropical Signalgrass (Urochloa Subquadripara) in Turf
Creator:
Pearsaul, David G
Place of Publication:
[Gainesville, Fla.]
Florida
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University of Florida
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english
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1 online resource (51 p.)

Thesis/Dissertation Information

Degree:
Master's ( M.S.)
Degree Grantor:
University of Florida
Degree Disciplines:
Agronomy
Committee Chair:
LEON-GONZALEZ,RAMON G
Committee Co-Chair:
SELLERS,BRENT ALAN
Committee Members:
SILVEIRA,MARIA LUCIA
ODERO,DENNIS C

Subjects

Subjects / Keywords:
bermudagrass -- brasu -- iwm -- verticutting
Agronomy -- Dissertations, Academic -- UF
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bibliography ( marcgt )
theses ( marcgt )
government publication (state, provincial, terriorial, dependent) ( marcgt )
born-digital ( sobekcm )
Electronic Thesis or Dissertation
Agronomy thesis, M.S.

Notes

Abstract:
Tropical signalgrass (TSG) has become a troublesome weed in south Florida following the removal of monosodium methane arsenate (MSMA), an effective and economical postemergence (POST) herbicide, from the turfgrass market at the end of 2012. Turfgrass managers have relied heavily upon this herbicide to control TSG, as well as many other weeds, for many years and are now searching for a comparable replacement. In an attempt to avoid a similar situation in the future, this thesis focuses on identifying an effective integrated weed management (IWM) strategy aimed at controlling TSG in bermudagrass turf. In one study, verticutting, a cultural practice that is regularly employed on many golf courses, was combined with preemergence (PRE) and POST herbicide treatments to determine whether TSG control would be increased using this integrated approach. The POST herbicides were then evaluated to determine the most efficacious combinations. Additionally, a PRE herbicide study was conducted to demonstrate the impact of seedling recruitment on TSG reestablishment. In this research, verticutting was found to provide limited weed control by itself. Amicarbazone demonstrated the ability to enhance other POST herbicides through synergistic activity on TSG control with multiple modes of action. PRE herbicides proved to be a crucial component of a TSG management strategy, with indaziflam showing the greatest level of efficacy of those herbicides tested. ( en )
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In the series University of Florida Digital Collections.
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Includes vita.
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Includes bibliographical references.
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Description based on online resource; title from PDF title page.
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This bibliographic record is available under the Creative Commons CC0 public domain dedication. The University of Florida Libraries, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.
Thesis:
Thesis (M.S.)--University of Florida, 2017.
Local:
Adviser: LEON-GONZALEZ,RAMON G.
Local:
Co-adviser: SELLERS,BRENT ALAN.
Electronic Access:
RESTRICTED TO UF STUDENTS, STAFF, FACULTY, AND ON-CAMPUS USE UNTIL 2018-06-30
Statement of Responsibility:
by David G Pearsaul.

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UFRGP
Rights Management:
Applicable rights reserved.
Embargo Date:
6/30/2018
Classification:
LD1780 2017 ( lcc )

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INTEGRATED CONTROL OF TROPICAL SIGNALGRASS ( Urochloa s ubquadripara ) IN TURF By DAVID PEARSAUL 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 2017

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2017 David Pearsaul

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To my m om

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4 ACKNOWLEDGMENTS I would like to thank my major advisor, Dr. Ramon Leon, for giving me this opportunity and making this an enjoyable learning experience. I also thank my committee members, Dr. Maria Silveira, Dr. Brent Sellers, and Dr. Calvin Odero for thei r support and guidance throughout this process. I would also like to thank Ryan Duffell and Chuck Calhoun at Sandridge Golf Club and Christian Millican at Pointe West Country Club for being so accommodating over the past few years. I graciously thank Synge nta for their support, as this would not have been possible otherwise. Above all, I would like to thank my family, my wife Samantha for her unwavering support and our son Henry for motivating me above a ll else.

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5 TABLE OF CONTENTS page ACKNOWLEDGMENTS ................................ ................................ ................................ .. 4 LIST OF TABLES ................................ ................................ ................................ ............ 6 LIST OF FIGURES ................................ ................................ ................................ .......... 7 ABSTRACT ................................ ................................ ................................ ..................... 8 CHAPTER 1 INTRODUCTION ................................ ................................ ................................ .... 10 Impact of Weeds ................................ ................................ ................................ ..... 10 MSMA Ban ................................ ................................ ................................ .............. 12 Weeds and Turf Quality ................................ ................................ .......................... 13 Integrated Weed Management ................................ ................................ ............... 14 Nonchemical Control ................................ ................................ ........................ 14 Chemical Control ................................ ................................ .............................. 15 2 AN INTEGRATED APPROACH TO TROPICAL SIGNALGRASS ( Urochloa subquadripara ) CONTROL IN TURF ................................ ................................ ...... 19 Summary ................................ ................................ ................................ ................ 19 Background ................................ ................................ ................................ ............. 20 Materials and Methods ................................ ................................ ............................ 23 Results and Discussion ................................ ................................ ........................... 25 IWM Study ................................ ................................ ................................ ........ 25 POST Study ................................ ................................ ................................ ..... 28 3 THE IMPACT OF SEEDLING RECRUITMENT ON TROPICAL SIGNALGRASS ( Urochloa subquadripara ) REESTABLISHMENT IN TURF ................................ .... 3 7 Summary ................................ ................................ ................................ ................ 3 7 Background ................................ ................................ ................................ ............. 3 7 Materials and Methods ................................ ................................ ............................ 3 9 Results and Discussion ................................ ................................ ........................... 40 4 SUMMARY AND CONCLUSIONS ................................ ................................ .......... 4 4 LIST OF REFERENCES ................................ ................................ ............................... 4 7 BIOGRAPHICAL SKETCH ................................ ................................ ............................ 5 1

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6 LIST OF TABLES Table page 2 1 Herbicides, formulations, and rates used in the experiments ............................. 30 2 2 Effects of verticutting, preemergence (PRE) herbicide, and postemergence (POST) herbicide on tropical signalgrass (TSG) cover at 2, 4, 8, and 12 wk after initial treatment (WAIT) ................................ ................................ ............... 31 2 3 Interaction (P<0.0001) between preemergence (PRE) and postemergence (POST) herbicides on tropical signalgrass (TSG) cover at 52 wk after initial treatment (WAIT) ................................ ................................ ................................ 32 2 4 Postemergence (POST) tropical signalgrass (TSG) % visual cover and % of squares ot of 36 occupied by tropical signalgrass (% grid) at 2, 4, 8, and 12 wk after initial treatment (WAIT) for site 1 ................................ ........................... 33 2 5 Postemergence (POST) tropical signalgrass (TSG) % visual cover and % of squares ot of 36 occupied by tropical signalgrass (% grid) at 2, 4, 8, and 12 wk after initial treatment (WAIT) for site 2 ................................ ........................... 34 2 6 Synergism between amicarbazone and multiple mix partners at 2, 4, 8, and 12 wk after initial treatment (WAIT) ................................ ................................ .... 35 2 7 Turfgrass injury observed in postemergence (POST) herbicide study at 2, 4, and 8 wk after initial treatment (WAIT) ................................ ............................... 36

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7 LIST OF FIGURES Figure page 3 1 Tropical signalgrass seedling emergence over time in the nontreated control and areas treated with S metolachlor and indaziflam. Weeks after treatment (WAT). Error bars correspond to standard error of mean ................................ ... 42

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8 Abstract of Thesis Presented to the Gradu ate School of the University of Florida in Partial Fulfillment of the Requirements for the Degree of Master of Science INTEGRATED CONTROL OF TROPICAL SIGNALGRASS ( Urochloa subquadripara ) IN TURF By David Pearsaul December 2017 Chair: Ramon Leon Major: Agronomy Tropical signalgrass (TSG) has become a troublesome weed in south Florida following the removal of monosodium methane arsenate (MSMA), an effective and economical postemergence (POST) herbicide, from the turfgrass market at the end of 2012. Turfgrass managers have relied heavily upon this herbicide to control TSG, as well as many other weeds, for many years and are now searching for a comparable replacement. In an attempt to avoid a similar situation in the future this thesis fo cuses on identifying an effective integrated weed management (IWM) strategy aimed at controlling TSG in bermudagrass turf. In one study, verticutting, a cultural practice that is regularly employed on many golf courses, was combined with preemergence (PRE) and POST herbicide treatments to determine whether TSG control would be increased using this integrated approach. The POST herbicides were then evaluated to determine the most efficacious combinations. Additionally, a PRE herbicide study was conducted to demonstrate the impact of seedling recruitment on TSG reestablishment. In this research, verticutting was found to provide limited weed control by itself. Amicarbazone demonstrated the ability to enhance other POST herbicides through

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9 synergistic activity o n TSG control w ith multiple modes of action. PRE herbicide s proved to be a crucial component of a TSG management strategy, with indaziflam showing the greatest level of efficacy of those herbicides tested.

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10 CHAPTER 1 I NTRODUCTION Optimal turfgrass quality is the goal of many golf course superintendents and sod farm managers. Turfgrass managers constantly deal with both biotic and abiotic stressors while attempting to maintain ideal playing conditions or grow high qualit y sod. Diseases, insects, and weeds are the most common biotic agents reducing turf quality and growth while water, nutrient deficiencies and temperature extremes are important abiotic stresses (Taiz et al. 2015). All sources of stress must be considered w hen implementing a turf management strategy. An unhealthy plant is more susceptible to injury from diseases or insects and, in the case of turfgrass, a thinned canopy can favor unwanted weed pressure ultimately eliminating the competitive advantage that th e turfgrass may have had originally. This is similar to the agronomic cropping practice of increasing plant density and decreasing the spacing between rows in order to reduce the area and resources available to weeds (Buhler 2002). Impact of Weeds Weeds pr esent an enormous challenge for agriculture as well as turfgrass production and management as evidenced by the estimated losses in corn and soybean yields in the absence of any contro l measures. Soltani et al. (2016, 2017 ) reported that approximately 50% of corn and soybean yields in North America (valued at $27 billion and $16 billion, respectively) would be lost annually by uncontrolled weeds. In 2012, the United States Environmental Protection Agency (USEPA) estimated the world market value for herbici des and plant growth regulators (PGR) to be approximately $25 billion, accounting for nearly half of all pesticide sales in the world (Atwood and Paisley Jones 2017). Chemical control is the most heavily relied upon

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11 method of pest management primarily beca use it is effective. In the turfgrass industry, it allows for selective removal of undesired plants without harming the desired species a process that can be especially difficult when the weed is a grass species as well. Herbicide metabolism, or breakdown to non toxic compounds, forms the basis for turfgrass selectivity compared to susceptible weeds that are less able to metabolize selective herbicides (Cole 1994). Heavy reliance upon herbicides has led to additional problems most notably herbicide resist ance by weeds Herbicide resistance is defined by the Weed Science of any herb icide may eventually select for resistance within a population of weeds. The herbicide resistant biotypes will then pass this ability on to their progeny and the population will experience a shift towards a higher frequency of resistant plants. Although he rbicide resistance in agricultural cropping systems has been occurring for several decades (Ryan 1970), Brosnan and Breeden (2013) reference s everal studies confirming a more recent reporting of herbicide resistance in turfgrass. These include resistant bi otypes of annual bluegrass ( Poa annua L. ), goosegrass and smooth crabgrass ( Digitaria ischaemum (Schreb.) Schreb. Ex Muhl ). Misuse of an herbicide can also affect other similar herbicides as well. Cutulle et al. (2009) reported prodiamine resistant annual bluegrass biotypes that also displayed reduced sensitivity to pendimethalin, another preemergence (PRE) herbicide with the same mode of action. Weed adaptability can also occur in response to nonchemical selection pressures. Barrett (1983) illustrated the consequence of overusing nonchemical control

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12 methods. For example, through the process of frequent and intensive hand weeding in rice ( Oryza sativa L.) fields, Echinochloa crus galli (L.) Beauv. var. oryzicola (Vasing Ohwi) has evolved morphology traits t hat make it look more similar to rice than to its close weedy relative, Echinochloa crus galli (L.) Beauv. var. crus galli This serves as a reminder that any control method, chemical or not, can lose efficacy due to weed adaptations especially if used imp roperly, repeatedly, and without integration with other forms of control. MSMA Ban Another negative consequence of relying heavily on a single method of control was made evident when monosodium methane arsonate (MSMA) was banned from use on golf course s an d sod farms in Florida. This herbicide was an affordable and highly effective postemergence (POST) herbicide used to control many troublesome weed species. MSMA contains an organic form of arsenic which is relatively nontoxic, less so than even aspirin whe n considering acute toxicity (Brosnan and Breeden 2014). Inorganic arsenic, however, is highly toxic and public concern arose around whether or not the organic arsenic from MSMA could be converted to its inorganic form in the soil and leach into groundwate r or run off into surface water. In a lysimeter study under greenhouse conditions, Mahoney et al. (2015) documented that after MSMA applications to bermudagrass, a small fraction (<10%) of the herbicide was quickly converted to As(III), but after 8 weeks, more than 80% of the arsenic present in pore water was in the inorganic form As(V). However, Matteson et al. (2014) conducted year long experiments on managed turfgrass systems under field conditions to determine the fate of arsenic after applications of M SMA. The authors demonstrated that a majority of arsenic remained in the turfgrass foliage and soils, while the concentrations of dissolved

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13 arsenic found in pore water measured at a depth of 76 cm was indistinguishable from background concentrations. They concluded that, following normal applications of MSMA, minimal leaching of arsenic into groundwater would occur, however, repeated applications could pose additional risks to the environment. Ultimately, the USEPA ruled that MSMA could no longer be sold fo r use on golf courses or sod farms after December 31, 2012. Turfgrass managers were left without a reliable and affordable means of controlling grass weeds such as tropical signalgrass crabgrass, goosegrass, and dallisgrass ( Paspalum dilatatum Poi.) and, as a result, many infestations were allowed to grow unchecked. Weeds and Turf Quality Turfgrass quality is dependent upon density, uniformity, and color, all of which can be disturbed by the presence of weeds. Noticeable differences in leaf blade width and color, as well as undesirable seedhead production, are some of the ways that weeds can negatively affect turfgrass aesthetics. Weeds have evolved to become excellent competitors for space and resources. Some weeds, such as goosegrass [ Eleusine indica (L.) Gaertn.] can produce up to 140,000 seeds plant 1 (Chin 1979) and are well adapted for subtropical conditions such as those found in Florida (Chauhan and Johnson 2008). Tropical signalgrass (TSG), also known as smallflowered alexandergrass, two finge rgrass, and two spiked panic (Speedy 2002), is a perennial grass species that is native to Asia and Australia and is now found in Florida, Africa, and Mexico among other places (Murphy et al. 1992). Morphological characteristics include 0.75 to 1.25 cm wid e leaf blades that can be up to 2 cm long and flowering (Unruh et al. 2009). TSG is capable of reproducing by seed and stolons and can be

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14 found on golf courses, athletic fi elds, sod farms, and lawns. This weed forms dense m ats on golf course fairways and in recently harvested bare areas of sod farms. Due t o a lack of killing freezes in s outh Florida, this weed is able to grow as a perennial and increase its footprint year af ter year. Integrated Weed Management Integrated weed management (IWM) combines nonchemical control methods, such as cultural, mechanical, and biological practices, with traditional chemical control in an attempt to avoid resistance and reduce the heavy rel iance upon chemicals to perations (Danyal et al. 2008). Nonchemical Control Cultural practices on golf courses are done to promote ideal growing conditions in an attempt to increase and/or maintain overall health, aesthetics, and playability of the turfgrass. Cultural practices routinely implemented on the golf course include top dressin g, aeration, verti cutting, water management, and fertilization. These practices could all be easily integrated into an IWM strategy if they provide additional weed control at little to no additional cost. Top dressing is usually a weekly or biweekly process of spreading sand across a putting green to smooth and firm up the surface. Aeration involves removing small cores across the entire surface of the turf in an attempt to improve water and nutrient movement throughout the soil a s well as relieve compaction that takes place from constant walking and/or driving. Verticutting is the process of chopping up stolons, which are horizontal stems that grow aboveground and take root at certain intervals forming new plants, and removing exc ess thatch using

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15 vertical blades that cut down into the turf. This practice promotes new growth and healthy, dense turfgrass. For densely growing weeds such as TSG and crabgrass, verticutting may eliminate apical dominance and encourage bud sprouting (Robe rtson et al. 1989). Apical dominance inhibits lateral bud growth, and by breaking this signaling pathway the plant will respond by sprouting buds at axillary meristems which will exhaust carbohydrate reserves stored in the shortened stolons and leave the p lant more susceptible to biotic or abiotic stressors (Vengris 1962). Verticutting may also initiate weed seedling emergence by opening space in the turf canopy (Johnson 1979), a result that may also be beneficial if an herbicide applicat ion is forthcoming to target susceptible seedlings. Verticutting is done at most golf courses multiple times throughout the year, making this an ideal cultural practice to implement into an IWM strategy. If combined with an herbicide that is safe to the turfgrass, this proce ss could aid in weed control while also encouraging turfgrass regrowth and decreasing the time required for reestablishment into the vacated areas. Chemical Control Chemical control methods include PRE and POST herbicides each of which have multiple modes of action that can be used in combinations to avoid selecting for herbicide resistant weed biotypes. Current chemical control options for some difficult to control grass weeds in turf are limited, especially for POST applications. The banning of MSMA has f orced some turfgrass managers to use non selective POST herbicides such as glyphosate to eradicate weeds and, in doing so, the desired turf species as well. Teuton et al. (2004 a ) reported >90% TSG control in the greenhouse from early POST (2 to 8 leaf sta ge) applications of herbicides such as asulam, trifloxysulfuron, imazaquin, and metsulfuron. However, these results could not be duplicated in the field

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16 against mature stands of TSG, similar to what turfgrass managers are faced with after a year or two of ineffective POST herbicide applications following the banning of MSMA. More recently, Cross et al. (2016) reported excellent (>97%) TSG control 12 wk after initial treatment (WAIT) from POST fall applications of amicarbazone with rates up to 0.49 kg ai ha 1 Amicarbazone proved to be a valuable tank mix partner with numerous other herbicides, however, this may not be an economically feasible option at the rates being tested (i.e. more than $1000 per acre per treatment). Additionally, summer applications may be more advantageous than fall applications for allowing the turfgrass to recover and reestablish into the bare areas left after TSG elimination before turf growth decreases during fall and winter months. Teuton et al. (2004 a ) also examined PRE herbicide options for controlling this weed in sod farms. Certain treatments, such as imazapic + 2,4 D, performed extremely well (>90% TSG control) across multiple sites over two years. POST herbicides have been the predominant tool used for TSG control on golf cour ses. MSMA was an affordable, effective herbicide that controlled many grass weeds in turf. Amicarbazone is a photosystem II (PSII) inhibitor that can be taken up by roots and result in chlorosis, stunting, and eventual necrosis of leaf tissue (Dayan et al. 2009). This herbicide is generally safe on cool and warm season turfgrasses and has activity on both broadleaf and grass weeds (Anonymous 2014). Mesotrione is a 4 hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor whose symptomology includes bleaching of leaf tissue, which is a result of insufficient carotenoid production (Armel et al. 2007). Trifloxysulfuron and thiencarbazone + foramsulfuron + halosulfuron are all acetolactate synthase (ALS) inhibitors capable of being translocated to the growing

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17 region s of plants resulting in chlorosis and leaf tissue stunting (Shaner 2014). Although these POST herbicides are commonly used alone, in combination they could provide a to assimilate photosynthates, and 3) meristematic regions through extensive translocation within the plant. Vegetative propagation is a dominant method of establishme nt for many turfgrass weeds, however, seedling recruitment also plays an important role and must be addressed in management plans. PRE herbicides are effective tools in turfgrass weed management. They are capable of inhibiting seedling shoot and/or root gr owth and can affect the susceptible meristematic regions of emerging seedlings as they penetrate the soil surface (e.g. chloracetimides such as S metolachlor). Some PRE herbicides, such as the dinitroanaline (DNA) compounds, are capable of root pruning and growth inhibition of mature species including turfgrass (Fishel and Coats 1993). Many PRE herbicides are applied at scheduled intervals throughout the year depending on projected germination timings of the targeted weed species such as crabgrass (Anonymou s 2012). Their application may also be beneficial following the eradication of patchy weeds via a POST herbicide application. Subsequent bare patches can invite a flush of germination if weed control is timed incorrectly. Indaziflam is a cellulose biosynt hesis inhibiting herbicide used primarily PRE in turfgrass (Henry et al. 2012), but has also shown the potential to enhance POST activity of 2,4 D, fluroxypyr, and simazine (McCullough et al. 2015). Additionally, at the time of publication, Brosnan and Bre eden (2013) reported that there were no known instances

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18 of cellulose biosynthesis inhibiting resistant biotypes present in any weed populations. The authors presumed there to be a reduced likelihood of resistance developing with this mode of action due to its multiple target sites within the corresponding biosynthetic pathway, which serves to increase the value of this herbicide. S metolachlor is a very long chain fatty acid (VLCFA) biosynthesis inhibitor that primarily targets seedlings as they emerge from the soil (Shaner 2014). This results in stunted, malformed seedlings that eventually succumb to necrosis. The present thesis evaluated verticutting along with PRE and POST herbicides in a series of studies to identify integrated strategies aimed at managing TSG in bermudagrass turf. There are currently few to no options available for controlling this weed in an effective and economical manner. In an effort to avoid developing herbicide resistance, as well as relying too heavily on a chemical control that may be taken away, the path forward should focus on an IWM approach that incorporates cultural, mechanical, biological, and/or chemical control methods.

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19 CHAPTER 2 AN INTEGRATED APPROACH TO TROPICAL SIGNALGRASS ( Urochloa subquadripara ) CONTROL IN TURF Summary Tropical signalgrass (TSG) is one of the most problematic weeds found on golf courses, s ports fields, and sod farms in s outh Florida. The r ecent ban of monosodium methane arsonate (MSMA), an organic arsenical herbicide, from urban areas in Florida has left turfgrass managers searching for effective management options. In an effort to avoid relying solely on postemergence (POST) chemical control, this research examined the effect of combining a cultural practice, verticutting, along with preemergence (PRE) and POST herbicides as an integrated weed management (IWM) approach to controlling tropical signalgrass in hybrid bermudagrass. Field experiments were conducted at multip le locations over two years in s outh Florida to: (1) determine whether verticut ting prior to herbicide applications increases TSG control and (2) identify herbicide programs that effectively control TSG. No interactions between verticutting and herbicide programs were detected, but verticutting consistently provided a slight reductio n (~10%) in TSG cover. Treatments containing a PRE herbicide resulted in a significant reduction (20 50%) in TSG cover at 52 wk after initial treatment (WAIT), while some POST herbicide treatments reduced TSG cover to <20% at 52 WAIT. A study was conducted to determine which POST herbicide combinations were most and 12 WAIT, but synergistic responses were observed between amicarbazone and mesotrione, trifloxysulfuron, and thie ncarbazone + foramsulfuron + halosulfuron. Two and three way combinations of amicarbazone with these POST herbicides resulted in 80% TSG control at 4, 8, and 12 WAIT, with some reaching 100% TSG control at 4

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20 WAIT. Based on these data, verticutting may pr ovide limited complementary control but certain combinations of POST herbicides exhibited excellent (>95%) TSG control. Background Tropical signalgrass is a perennial grass species capable of reproducing by seed and stolons and is commonly found in lawns, cultivated fields and disturbed areas (Murphy et al. 1992). Its dense, aggressive growth habit allows for quick colonization of any bare patches or areas where turfgrass density is low. This species gets its name from the angle of branching of its seedhead al. 1992). Other common names include smallflowered alexandergrass, green summergrass, two spiked panic, and two fingergrass (Speedy 2002). It is native to Asia and Australia (Murphy et al. 1992) and now occurs i n Florida, Maryland, Hawaii, and Puerto Rico (USDA 2002). TSG has become one of the most troubl esome weeds on golf courses in s outh Florida. This weed thrives in tropical conditions, with seed germination (Teuton et al. 2004a) and vegetative growth being optimal around 25 C and in moist soil conditions. Frost and killing freezes are uncommon in s outh Florida, thus TSG has the potential to grow year round. Monosodium methane arsonate (MSMA), the standard for chemical control of this weed, was banned for ur ban uses including turfgrass in 2012, and any remaining product in storage after this time could be applied until December 31, 2013 (Brosnan et al. 2009). This has created a need to find alternative effective control tools with the ultimate goal of develop ing an integrated approach for managing this weed. Prior studies have looked at herbicide use as the sole method for TSG control. TSG plants at 8 wk after initial treatmen t (WAIT) for all summer applied postemergence

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21 (POST) herbicides and herbicide combinations that were tested, including asulam, ethofumesate, and quinclorac. Better results were observed with bare ground preemergence (PRE) herbicide applications following b urndown with glyphosate, al though efficacy decreased 5 WAIT. Integrated herbicide management utilizes PRE herbicides, POST herbicides, tank mixes, and differing modes of action. This alone does not constitute an integrated weed management (IWM) program. Ta nk mixing herbicides with different modes of action can be effective, but to truly implement an IWM approach there must also be an (2008) cite the importance of accounting f or cropping system design to allow for easier integration of IWM strategies into farming operations. Common cultural practices that could easily be implemented by turfgrass managers into IWM strategies include mowing, irrigation, fertilization, and cultiva tion. Bergkvist et al. (2017) showed that rhizome fragmentation of quackgrass ( Elymus repens (L.) Gould) in the early summer reduced rhizome biomass by up to 60%. In turfgrass management, verticutting is a cultural practice that uses vertical mower blades to cut down into the turf removing excess thatch and fragmenting stolons, which promotes new growth and healthy turfgrass. We hypothesized that verticutting several days before applying the herbicide will result in an increase in TSG control by weakening T SG underground reserves while favoring turf growth. There are several POST herbicides available that have shown activity on TSG, however, they require high rates and multiple sequential applications to provide acceptable control while also potentially harm ing the turfgrass (McCarty and Estes

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22 2014). Amicarbazone (AMI) is a photosystem II (PSII) inhibitor that is active in the soil and can be taken up by the roots of TSG plants recovering from foliar applied herbicides. Symptoms include chlorosis, stunting an d eventual necrosis of leaf tissue (Dayan et al. 2009). This herbicide has been shown to be safe on multiple cool and warm season turfgrasses and is labeled for control of various broadleaf weeds as well as some grass es such as annual bluegrass ( Poa annua L.) (Anonymous 2014). Mesotrione (MESO) is a 4 hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor that causes bleaching of the leaf tissue, and subsequent necrosis, resulting from insufficient carotenoid production (Armel et al. 2007). Trifloxysulfuron (T SS) and thiencarbazone + foramsulfuron + halosulfuron (TFH) are all acetolactate synthase (ALS) inhibitors that can be extensively translocated to growing regions and result in chlorosis and stunting of leaf tissue (Shaner 2014). We hypothesized that a POS T herbicide program utilizing a three tier approach: (1) above ground bleaching and burn down of leaf tissue by mesotrione, (2) below ground root uptake of amicarbazone to inhibit any new growth, and (3) systemic activity of an ALS inhibiting herbicide to target meristematic growth, could increase TSG control while allowing the use of lower rates and fewer applications. Although managers predominantly focus their efforts on controlling established TSG plants, we hypothesized that a PRE herbicide with strong residual properties might be necessary to effectively reduce TSG populations from one season to the next. This is true of many weed management approaches, however, little research has been done identifying the role of seedling recruitment in TSG regrowth after POST control actions. The importance of implementing an IWM strategy and avoiding sole reliance on POST chemical control has been highlighted by the situation that now faces many

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23 turfgrass managers as they attempt to combat TSG infestations. There is a need to further explore the role that cultural practices can play as part of an integrated management program. Recent research has shown acceptable levels of TSG control using POST herbicides such as amicarbazone and thiencarbazone + foramsulfuron + hal osulfuron (McCarty and Estes 2014; Cross et al. 2016). However, as was the case with MSMA, if a chemical is banned then turfgrass managers are left without any effective tools at their disposal. Therefore, the objectives of this research were to: (1) deter mine whether verticutting prior to herbicide applications increases TSG control and (2) identify herbicide programs that effectively control TSG. Materials and Methods Field experiments were conducted in 2015 and 2016 in Vero Beach, FL at Sandridge Golf Cl ub and Pointe West Country Club to determine the effects of verticutting, PRE herbicides, and POST herbicides (Table 2 1) on TSG control. The IWM study was conducted at both sites in 2015 and one site (Sandridge) in 2016 for a total of three sites/years. T he POST study was conducted at both sites in 2016. S oil type was a Myakka fine sand at both locations Experiments were conducted in areas with rough cover). The IWM study was conducted to evaluate the benefits of integrating different t ools for TSG control. For this a split plot factorial with four replications was used to evaluate the integration of three factors: verticutting (with and without), PRE (with indaziflam and without), and POST herbicides (AMI, MESO, TSS, TFH, and nontreated control). The main plot was verticutting and PRE and POST herbicide combinations were the subplots. Verticutting was performed 14 days before initial herbicide treatment

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24 (DBIT) using a Toro triplex gree ns mower (Greensmaster Triflex Hybrid 3320, Toro, Bloomington, MN) with verticutting blade attachments. Two passes, at a depth of 2.5 cm, were made in opposite directions to increase the amount of stolon fragmentation. Two POST herbicide applications were made with a 21 d interval, and a PRE herbicide application was made with the second POST herbicide application. The POST study was conducted to identify the most effective POST herbicide combinations. This study was arranged as a randomized complete block design with four replications. Two POST herbicide applications were made with a 21 d interval. For both studies, experimental units were 2.25 m 2 (1.5 m by 1.5 m), herbicide treatments were applied in a water carrier volume of 374 L ha 1 and all applications were made with a CO 2 pressurized boom sprayer equipped with three 8002VS flat fan nozzles (TeeJet Spraying Systems Co., Wheaton, IL) on 45 cm spacing. Visual evaluations were recorded weekly beginning 3 DBIT and ending 52 WAIT. TSG cover was e valuated visually on a 0 to 100% scale, and also using a quadrat (1 m by 1 m) with a grid to record the total number of units out of 36 that contained TSG. Weed control was evaluated visually on a 0 to 100% scale (0% = no reduction in weed population and n o symptoms of injury, 100% = complete elimination of weed populations or only dead weed tissue present). Bermudagrass density was visually estimated on a 0 to 100% scale and turf color and quality were evaluated visually on a 1 to 9 index scale (1 = dead t urf; 6 = acceptable turf quality/color; 9 = excellent turf quality/color) (McCarty et al. 1991). Bermudagrass injury (phytotoxicity) was visually estimated on a 0 to 100% scale (0% = no injury; 100% = complete elimination of turfgrass or only dead tissue p resent).

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25 For the IWM study, nontransformed data were subjected to ANOVA using PROC GLIMMIX in SAS (SAS Institute, Cary, NC) after confirming data normality and homoscedasticity. An ANOVA was performed for each parameter by assessment date because an interaction was present between assessment date and other factors (P<0.01). Site by POST treatment interactions were significant (P<0.0001), but data was pooled due to the absence of crossover interactions. For the POST herbicide study, square root transfo rmed grid data, arcsine transformed visual cover data, and nontransformed control data were all subjected to ANOVA by PROC GLIMMIX in SAS. To identify the presence of synergism, the Colby (1967) equation was used to calculate expected means for the herbici de combinations. Expected means were then compared to the observed means to determine whether the combinations were synergistic, 0.05) test. Results and Discussion IWM Study Verticutting did not increase the efficacy of POST herbicide applications and interactions between these two factors were not significant (P>0.16). V erticutting did provide a small but consistent (P<0.0001) red uction in TSG visual cover through 12 WAIT (Table 2 2 ) however, long term control with verticutting alone will be minimal Busey and Johnston (2006) also demonstrated that cultural practices alone (e.g. mowing, fertilization, irrigation) may not provide a cceptable, long term results. They found that after a 3 yr period without herbicide use, most plots exhibited poor turf quality and high levels of weed pressure. The removal of photosynthetic tissue and fragmentation of stolons resulting from verticutting might reduce the amount of

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26 carbohydrate reserves each individual TSG plant can access for regrowth and recovery. At 2, 4, 8, and 12 WAIT, plots that were verticut had 39, 26, 29, and 36% TSG cover, respectively, while those that did not had 48, 34, 37, an d 43% TSG cover, respectively. Therefore, verticutting reduced TSG cover on average about 8%. IWM utilizes multiple tactics, some of which may provide very little control on their own, in a complementary manner. Most importantly, these tactics must be easi existing program (Swanton et al. 2008). Verticutting results in more active growing points, potentially increasing the number of vegetative buds that are killed by herbicides with translocation and systemic activity. Althou gh verticutting did not provide an increase in POST herbicide efficacy, future research examining this cultural practice with reduced rates of the effective POST herbicides tested in this study may prove beneficial in scenarios where verticutting is normal ly used. At 52 WAIT, there was an interaction between PRE and POST herbicide (P<0.0001; Table 2 3 ). S pecific combinations of PRE and POST herbicides increased long term control considerably. For example, amicarbazone + thiencarbazone + foramsulfuron + halo sulfuron treatment benefited the most from the PRE applications exhibiting 73% TSG cover without PRE and 25% with PRE. TSG seeds exhibit a dormancy mechanism that is still not well understood (Teuton et al. 2004a), but our results indicate that germination can occur during the summer and fall. Therefore, PRE herbicides should complement POST applications to prevent new seedling recruitment. Indaziflam, a cellulose biosynthesis inhibitor, is an effective PRE herbicide used in turfgrass (Henry et al. 2012) an d has shown the ability to enhance POST activity of some herbicides such as 2,4 D, fluroxypyr, and simazine (McCullough et al. 2015) as

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27 well as provide PRE and early POST activity when applied alone to annual bluegrass (Brosnan et al. 2012). Although veget ative propagation of TSG seems to be the primary method of encroachment this research confirms the need for PRE herbicides in a management plan. POST herbicide treatments reduced TSG populations at all assessment timings (P<0.0001; Table 2 2). Amicarbazone + mesotrione + trifloxysulfuron provided the greatest reduction in TSG cover at 4, 8, and 12 WAIT resulting in 4, 4, and 7% TSG cover, respectively (nontreated check had 61, 70, and 74 % TSG cover at those timings). Other POST herbicide treatments provide d varying levels of control. The combination of amicarbazone + mesotrione + trifloxysulfuron includes three different modes of action, two of which (PSII and HPPD inhibitors) have been shown to produce synergistic responses in certain situations, primaril y against broadleaf weeds (Abendroth et al. 2006; Hugie et al. 2008). The other mode of action, ALS inhibition, has been shown to increase efficacy when tank mixed with amicarbazone (Cross et al. 2016). All POST herbicide treatments resulted in acceptable (<20%) levels of phytotoxicity to the bermudagrass turf at all assessment timings with the highest level of injury (18%) occurring in the amicarbazone + mesotrione + trifloxysulfuron treatments at 4 WAIT. Because these studies were performed on rough heigh t turf of lesser quality, further studies should be done to determine safety levels of these POST herbicides on well maintained, fairway height bermudagrass.

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28 POST Study POST herbicides were assessed individually and in all possible combinations to determi ne the most effective treatments. There was a significant (P<0.0001) interaction between site and treatment effects so data was analyzed separately. At Pointe West a ll treatments containing only one herbicide resulted in >70% TSG cover at all assessment t imings, with the exception of amicarbazone at 4 WAIT which contained 36% TSG cover (Table 2 4). Of the two way combinations, only those contain ing amicarbazone resulted in <26 % TSG cover at 4, 8, and 12 WAIT for both locations (Tables 2 4 and 2 5) The two way combinations without amicarbazone resulted in >65% TSG cover at all assessment timings at Pointe West and >50% at 8 and 12 WAIT at Sandridge Similarly, all three way combinations containing amicarbazone greatly outperformed the treatment that did not contain amicarbazone at 4, 8, and 12 WAIT (<22% versus >60% TSG cover, respectively at Pointe West and <10% versus >45% TSG cover, respectively, at Sandridge ). PSII inhibitors, such as amicarbazone, have shown synergistic responses when combined with HPP D inhibitors (Abendroth et al. 2006) and complementary responses when combined with ALS inhibiting herbicides (Cross et al. 2016). In this study, amicarbazone displayed potential synergistic responses for TSG control when combined with HPPD or ALS inhibit ors. Using the Colby (1967) equation, expected combination means were calculated and compared with the observed combination means to determine whether there was a synergistic response (Table 2 6 ). There was no interaction between site and treatment effect so data was combined. At 2 WAIT, amicarbazone and mesotrione provided 12% and 6% TSG control respectively resulting in an expected value of 17% TSG control. The observed value of the amicarbazone +

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29 mesotrione treatment was 71% TSG control, indicating a str ong likelihood of synergy. A similar trend was observed at multiple assessment dates and with various tank mix partners. All treatments containing amicarbazone resulted in unacceptable (>20%) levels of phytotoxicity at 4 WAIT with the four way herbicide co mbination causing >70% injury to the bermudagrass turf (Table 2 7 ). All treatments showed full recovery by 8 WAIT. Further studies are needed to confirm these levels of injury and investigate possible methods of mitigation. Our results demonstrated t hat the contribution of verticutting to an IWM strategy was considerably smaller when compared to the herbicide treatments. However, if the herbicide synergy exhibited by amicarbazone and multiple mix partners on TSG affects other weed species, amicarbazon e could be a useful tool for managing difficult to control weeds and herbicide resistance. As with many weed management strategies, a PRE herbicide was shown to be a necessary component by providing longevity of control of TSG indicating that seedling recr uitment is an important source of TSG population growth after POST applications. Verticutting may still provide a benefit to turfgrass managers if implemented into existing practices so as to not incur additional costs. The early summer application timing could al so be beneficial in s outh Florida by allowing for a longer period of time for bermudagrass to reestablish into the areas that were previously covered with TSG. Future research should look at reduced rates of these combinations in conjunction with v erticutting to see whether the minor effect of this cultural practice could be increased while reducing weed contro l cost and environmental impact.

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30 Table 2 1 Herbicides, formulations, and rates used in the experiments. Herbicide (abbreviation) Formulation Rate (g ai ha 1 ) Trade n ame Manufacturer Location amicarbazone (AMI) 70 WG 245 a Xonerate FMC Corporation Philadelphia, PA mesotrione (MESO) 4 SC 280 Tenacity Syngenta Crop Protection Greensboro, NC trifloxysulfuron (TSS) 75 WG 27.8 Monument Syngenta Crop Protection Greensboro, NC thiencarbazone + foramsulfuron + halosulfuron (TFH) b 60.5 WG 136 Tribute Total TM Bayer CropScience Research Triangle Park, NC Indaziflam (IND) 0.62 SL 32.7 Specticle Flo Bayer CropScience Research Triangle Park, NC a For the IWM study, AMI was applied at 147 g ai ha 1 when combined with other herbicides. b All treatments containing TFH received methylated seed oil at 1% v/v and ammonium sulfate at 1.5 lb A 1 All other treatments received a nonionic surfactant at 0.25% v/v.

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31 Table 2 2. Effects of verticutting, preemergence (PRE) herbicide, and postemergence (POST) herbicide s on tropical signalgrass (TSG) cover at 2, 4, 8 and 12 wk after initial treatment (WAIT). V isual G rid a Factors 2 WAIT 4 WAIT 8 WAIT 12 WAIT 2 WAIT 4 WAIT 8 WAIT 12 WAIT TSG cover (%) Verticutting No 48 a b 34 a 37 a 43 a 86 a 69 a 67 a 72 a Yes 39 b 26 b 29 b 36 b 72 b 53 b 53 b 56 b PRE No 44 31 36 44 a 81 64 64 a 69 a Yes 43 29 31 35 b 78 61 56 b 58 b POST No 58 a 61 a 70 a 74 a 86 89 a 89 a 89 a AMI c 45 b 31 b 37 b 46 b 83 72 b 69 b 75 b AMI + TFH d 43 b 23 c 22 c 31 c 75 61 b 58 b 64 b AMI + MESO e + TSS f 27 c 4 d 4 d 7 d 75 25 c 19 c 28 c ANOVA g Verticutting <0.0001 0.001 0.003 0.005 <0.0001 <0.0001 <0.0001 <0.0001 PRE 0.80 0.25 0.06 0.001 0.68 0.26 0.02 0.004 POST <0.0001 <0.0001 <0.0001 <0.0001 0.06 <0.0001 <0.0001 <0.0001 a Assessment was taken using a 1 m 2 grid dissected into 36 identical sections. Each section was assessed for the presence of TSG and the percent of sections containing TSG was calculated. b Values within columns and main factors with the same letter were not statistically different based on c Amicarbazone d Thiencarbazone + foramsulfuron + halosulfuron e Mesotrione f Trifloxysulfuron g There were no interactions between main factors.

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32 Table 2 3. Interaction (P<0.0001) between preemergence (PRE) and postemergence (POST) herbicides on tropical signalgrass (TSG) cover at 52 wk after initial treatment (WAIT) a V isual G rid b PRE POST No Yes No Yes % No 73 ab 85 a 90 a 100 a AMI c 75 a 51 b 97 a 75 a AMI + TFH d 73 ab 25 c 100 a 41 b AMI + MESO + TSS e 14 c 11 c 22 b 17 b a Data were collected from only one site/year due to renovations at one of the golf courses in 2017 that resulted in loss of the trial site. b Assessment was taken using a 1 m 2 grid dissected into 36 identical sections. Each section was assessed for the presence of T SG and the percent of sections containing TSG was calculated. c Amicarbazone d Thiencarbazone + foramsulfuron + halosulfuron e Trifloxysulfuron

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33 Table 2 4. Postemergence (POST) tropical signalgrass (TSG) % visual cover and % of squares out of 36 occupied by tropical signalgrass (% grid) at 2, 4, 8, and 12 wk afte r initial treatment (WAIT) at Pointe West Country Club Visual Grid a WAIT Treatment b 2 4 8 12 52 2 4 8 12 52 TSG cover (%) Check 88 a c 88 a 93 a 98 a 88 a 100 a 100 a 100 a 100 a 100 a AMI d 84 ab 36 b 76 abc 86 abc 85 ab 100 a 100 a 100 a 100 a 100 a MESO e 76 ab 74 a 85 abc 93 ab 83 ab 97 a 99 a 100 a 100 a 100 a TSS f 78 ab 79 a 86 ab 89 abc 86 a 99 a 100 a 100 a 100 a 100 a TFH g 83 ab 80 a 88 ab 91 abc 81 ab 99 a 100 a 100 a 100 a 100 a AMI+MESO 33 cd <1 c 14 d 23 d 61 a e 96 a 2 c 75 a 80 ab 99 a AMI+TSS 59 abc 10 bc 14 d 14 d 69 a d 98 a 90 a 65 ab 73 abc 99 a AMI+TFH 54 bcd 9 bc 10 d 14 d 51 b e 98 a 90 a 64 abc 79 ab 100 a MESO+TSS 74 ab 68a 75 abc 70 c 84 ab 96 a 100 a 99 a 99 a 100 a MESO+TFH 73 ab 76 a 79 abc 85 abc 86 a 99 a 100 a 100 a 100 a 100 a TSS+TFH 74 ab 71 a 73 bc 75 bc 73 abc 100 a 100 a 100 a 100 a 100 a AMI+MESO+TSS 31 cd 1 c 4 d 6 d 29 e 91 a 15 bc 24 cd 27 c 84 b AMI+MESO+TFH 18 d 0 c 14 d 21 d 36 de 94 a <1 c 73 a 95 a 94 ab AMI+TSS+TFH 29 cd 5 bc 5 d 7 d 64 a d 95 a 31 b 24 bcd 34 bc 99 a MESO+TSS+TFH 79 ab 73 a 61 c 69 c 80 ab 100 a 100 a 100 a 100 a 100 a AMI+MESO+TSS+TFH 12 d <1 c 2 d 7 d 39 cde 83 a <1 c 9 d 35 bc 99 a a Assessment was taken using a 1 m 2 grid dissected into 36 identical sections. Each section was assessed for the presence of TSG and the percent of sections containing TSG was calculated. b All treatments containing TFH received methylated seed oil at 1% v/v and ammonium sulfate at 1.5 lb A 1 All other treatments received a nonionic surfactant at 0.25% v/v. c d Amicarbazone e Mesotrione f Trifloxysulfuron g Thiencarbazone + foramsulfuron + halosulfuron

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34 Table 2 5 Postemergence (POST) tropical signalgrass (TSG) % visual cover and % of squares out of 36 occupied by tropical signalgrass (% grid) at 2, 4, 8, and 12 wk after initial treatment (WAIT) at Sandridge Golf Club Visual Grid a WAIT Treatment b 2 4 8 12 52 2 4 8 12 52 TSG cover (%) Check 58 a c 58 a 76 ab 79 ab 21 a 99 a 98 a 100 a 100 a 74 abc AMI d 54 ab 17 abc 38 cd 43 cd 14 ab 100 a 94 a 96 a 97 a 59 abc MESO e 55 ab 53 a 81 a 84 a 18 ab 97 a 98 a 100 a 100 a 72 ab TSS f 56 ab 50 abc 71 ab 75 ab 9 ab 100 a 100 a 100 a 100 a 50 abc TFH g 53 ab 49 abc 69 ab 71 ab 5 ab 98 a 99 a 100 a 97 a 34 abc AMI+MESO 11 cd <1 c 8 d 8 de 5 ab 63 ab 1 cd 29 b 32 b 18 abc AMI+TSS 10 cd 1 bc 3 d 4 e 2 b 54 a d 3 bcd 8 b 10 b 10 c AMI+TFH 25 bcd 25 abc 7 d 9 de 1 b 61 abc 26 b 26 b 29 b 11 bc MESO+TSS 58 a 56 a 65 a c 70 abc 18 ab 100 a 100 a 99 a 99 a 76 a MESO+TFH 54 ab 43 abc 59 abc 59 abc 14 ab 99 a 88 a 99 a 97 a 60 abc TSS+TFH 51 ab 31 abc 51 bc 56 bc 11 ab 99 a 99 a 97 a 99 a 51 abc AMI+MESO+TSS 4 d <1 c 5 d 3 e 1 b 16 d <1 d 15 b 13 b 7 c AMI+MESO+TFH 7 d < 1 c 8 d 6 e <1 b 38 bcd 4 bcd 15 b 14 b 5 c AMI+TSS+TFH 26 bcd 4 bc 5 d 6 e 2 b 98 a 22 bc 15 b 19 b 10 c MESO+TSS+TFH 44 abc 46 abc 61 abc 60 abc 5 ab 94 a 97 a 94 a 97 a 32 abc AMI+MESO+TSS+TFH 1 d 1 bc 4 d 5 e 2 b 17 cd 14 bcd 11 b 15 b 10 c a Assessment was taken using a 1 m 2 grid dissected into 36 identical sections. Each section was assessed for the presence of TSG and the percent of sections containing TSG was calculated. b All treatments containing TFH received methylated seed oil at 1% v/v and ammonium sulfate at 1.5 lb A 1 All other treatments received a nonionic surfactant at 0.25% v/v. c d Amicarbazone e Mesotrione f Trifloxysulfuron g Thiencarbazone + foramsulfuron + halosulfuron

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35 Table 2 6 Synergism between amicarbazone and multiple mix partners at 2, 4, 8, and 12 wk after initial treatment (WAIT). TSG control a Treatment b 2 WAIT 4 WAIT 8 WAIT 12 WAIT 52 WAIT c % AMI d 12 69 35 29 4 MESO e 6 31 4 2 3 TSS f 17 30 15 14 1 TFH g 22 30 16 13 5 AMI + MESO 71 (17) 100 (79) 85 (38) 80 (30) 30 (7) AMI + TSS 61 (27) 93 (78) 88 (45) 87 (39) 21 (5) AMI + TFH 59 (31) 80 (78) 88 (45) 85 (38) 44 (9) MESO + TSS 26 (22) 34 (52) 23 (18) 20 (16) 9 (4) MESO + TFH 15 (27) 36 (52) 28 (19) 17 (15) 5 (8) TSS + TFH 31 (35) 48 (51) 38 (29) 33 (25) 18 (6) a Numbers in parentheses represent the expected means generated using the Colby (1967) equation. If the expected is less than the actual, than the combinations resulted in a synergistic response. b Treatments containing TFH received methylated seed oil at 1% v/v and ammonium sulfate at 1.5 lb A 1 All other treatments received a nonionic surfactant at 0.25% v/v. c Data collected at only one site. d Amicarbazone e Mesotrione f Trifloxysulfuron g Thiencarbazone + foramsulfuron + halosulfuron

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36 Table 2 7 Turfgrass injury observed in postemergence (POST) herbicide study at 2, 4, and 8 wk after initial treatment (WAIT) P hytotoxicity Treatment a 2 WAIT 4 WAIT 8 WAIT % AMI b 0 27 def 0 MESO c 1 13 ef 0 TSS d 0 4 f 0 TFH e 1 3 f 0 AMI + MESO 6 60 abc 0 AMI + TSS 2 35 cde 0 AMI + TFH 0 42 bcd 0 MESO + TSS 1 2 f 0 MESO + TFH 4 8 f 0 TSS + TFH 0 3 f 0 AMI + MESO + TSS 8 64 ab 0 AMI + MESO + TFH 7 66 ab 0 AMI + TSS + TFH 6 45 bcd 0 MESO + TSS + TFH 3 22 def 0 AMI + MESO + TSS + TFH 6 74 a 0 a Treatments containing TFH received methylated seed oil at 1% v/v and ammonium sulfate at 1.5 lb A 1 All other treatments received a nonionic surfactant at 0.25% v/v. b Amicarbazone c Mesotrione d Trifloxysulfuron e Thiencarbazone + foramsulfuron + halosulfuron

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37 CHAPTER 3 THE IMPACT OF SEEDLING RECRUITMENT ON TROPICAL SIGNALGRASS (Urochloa subquadripara) REESTABLISHMENT IN TURF Summary Tropical signalgrass (TSG) has recently become one of the most difficult weeds to control on South Florida golf courses, athletic fields, and sod farms. This is partially due to th e banning of monosodium methane arsonate (MSMA), an organic arsenical herbicide that provided cost effective postemergence (POST) control of TSG. Since this ban, TSG populations have increased considerably, but it is not known how much of the population growth is due to seedling recruitment or vegetative propagation. A field experiment was conducted in South Florida to (1) determine the impact of seedling recruitment on TSG reestablishment and (2) evaluate S metolachlor and indaziflam for preemergence (PRE) TSG control. At 2, 4, and 7 wk after the PRE application, the nontreated control had 135, 538, and 581 TSG seedlings m 2 respectively. S metolachlor provided limited TSG control compared to the nontreated control, while indaziflam provided >70% reduction in TSG seedling emergence. Based on the high levels of TSG seedling emergence observe d, an effective PRE herbicide is an important component of TSG integrated management programs. Background Tropical signalgrass (TSG) is a perennial grass species that is adapted to environmental conditions in Florida. The germination of this grass weed req uires high soil moisture levels and an optimal temperature of 25 C with no apparent light resemblance to a signal flag through its angle of branching (Murphy et al. 1992), a nd other common names include smallflowered alexandergrass, green summergrass, two

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38 spiked panic, and two fingergrass (Speedy 2002). TSG can be found in intensively managed areas such as lawns, athletic fields, sod farms, and golf courses (Murphy et al. 199 2). Recently, TSG has become one of the most difficult weeds to manage on golf courses and sod farms in Florida, especially in the central and southern regions of the state. Two reasons for this distribution are first, the lack of killing freezes in this a rea, which would slow down the rate of TSG establishment and, second, the banning of monosodium methane arsonate (MSMA) for use in turfgrass in Florida. MSMA is an organic arsenical herbicide that had become the standard for chemical control of TSG, as wel l as many other weeds in turfgrass. After its banning in 2012, turfgrass managers were left without inexpensive, effective postemergence (POST) herbicides that were also safe to use on bermudagrass or St. Augustinegrass resulting in the establishment of la rge, dense mats of TSG in golf course roughs and fairways. TSG populations have become so problematic that some turfgrass managers have been forced to use non selective POST herbicides such as glyphosate to eliminate this troublesome weed causing serious i njury to the turfgrass. Preemergence (PRE) herbicides, such as S metolachlor and indaziflam, are useful tools for weed management programs in turfgrass. S metolachlor is a very long chain fatty acid (VLCFA) biosynthesis inhibitor, which is primarily absorb ed by emerging shoots although some root absorption may also occur (Shaner 2014). Indaziflam is a cellulose biosynthesis inhibitor that can be applied PRE, resulting in weed seedling growth inhibition, or early POST with injury manifesting as root clubbing or stunted

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39 growth (Shaner 2014). These two herbicides are widely used in turfgrass in Florida, but their activity on TSG seedling emergence is unknown. Currently, most programs aimed at TSG management are focused on control through POST herbicide applicat ions. This weed is capable of propagating by seed and stolons, however, little research has been done to determine the extent to which establishment after POST control. Ther efore, the objectives of this research were to: (1) determine the impact of seedling recruitment on TSG reestablishment and (2) evaluate the efficacy of S metolachlor and indaziflam for PRE TSG control. Materials and Methods A field experiment was conducte d in 2016 in Vero Beach, FL on a golf course to determine the impact of seedling recruitment on TSG reestablishment. The experiment was conducted in an area with rough with TSG (>75% ground cover). The study was arranged as a randomized complete block design with four replications and experimental units measuring 2.25 m 2 (1.5 m by 1.5 m). Two glyphosate (0.8 kg ae ha 1 Touchdown HiTech 5 SL, Syngenta Crop Protection, Greensboro, NC) applications were made 21 d apart beginning in April to eliminate established TSG and bermudagrass, so TSG seedling emergence could be properly quantified. PRE herbicide treatments of indaziflam (0.03 kg ai ha 1 Specticle FLO 0.62 SL, Bayer CropScience, Research Triangle Park, NC ) or S metolachlor (1.37 kg ai ha 1 Pennant Magnum 7.62 EC, Syngenta Crop Protection, Greensboro, NC) were included in the second application. Diquat (4.5 kg ai ha 1 Reward 2 SL, Syngenta Crop Protection, Greensboro, NC) was applied following each assess ment timing to eliminate

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40 emerged seedlings. Herbicide treatments were applied in a water carrier volume of 374 L ha 1 and all applications were made with a CO 2 pressurized boom sprayer equipped with three 8002VS flat fan nozzles (TeeJet Spraying Systems C o., Wheaton, IL) on 45 cm spacing. Visual evaluations were conducted at 1, 2, 4, 7, 10, and 14 wk after PRE application (WAT). TSG seedlings and shoots from stolons were counted in a 1 m 2 area of each experimental unit and analyzed as a cumulative count ov er the duration of the study. Nontransformed data were subjected to ANOVA using PROC GLIMMIX in SAS (SAS Institute, Cary, NC) after confirming data normality and homoscedasticity. Means Results and Discussion No differences in TSG shoot regrowth were detected among treatments. This h level of efficacy for TSG control. Cumulative TSG seedling counts were highest in the nontreate d control and S metolachlor, while indaziflam resulted in >70% fewer seedlings m 2 14 WAT (Figure 3 1). At 4 WAT, approximately 550 seedlings m 2 had emerged in both the nontreated control and S metolachlor plots while indaziflam resulted in nearly one thi rd of the seedling emergence. The prediction model generated by Teuton et al. (2005) estimates a late March or early April emergence date for TSG in Vero Beach, FL. However, in our case emergence initiated a few weeks after the prediction, and there was a noticeable peak in seedling emergence between 2 and 4 WAT, which corresponded to the end of May. This delayed emergence could be due to reductions in seed dormancy resulting from the changes in soil temperature after the elimination of TSG and bermudagrass canopies. Also, it is possible that early emerging seedlings were eliminated with the

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41 glyphosate application. By 14 WAT, there were nearly 800 seedlings m 2 in the S metolachlor treatment. This research confirmed the need for an effective PRE herbicide wh en managing TSG infestations. In situations of complete elimination of existing TSG plants, seedling recruitment will be a driving force for TSG reestablishment. The use of glyphosate or other non selective and highly injurious herbicide programs to elimin ate mats of TSG, as some turf managers have been forced to do, could allow for a situation of enhanced seedling recruitment as the desired turf species is recovering and unable to immediately reestablish into treated areas. A PRE herbicide would be importa nt in this scenario, assuming the herbicide does not have any adverse effects on the desired species such as root pruning or growth inhibition. Teuton et al. (2004b) demonstrated high levels of control (>90% in some instances) with metolachlor initially, h owever, efficacy was generally not sustained past 8 WAT. In the present study, S metolachlor performed similarly to the nontreated control at each assessment timing. The contrasting results between both studies suggest that environmental factors might play an important role for S metolachlor activity on TSG. Indaziflam is an effective PRE herbicide, as evidenced by the cessation in seedling emergence after 4 WAT, however, there are also some drawbacks that must be taken into consideration concerning turfgra ss inhibition. Brosnan et al. (2014) demonstrated an increase in days required to reach 50% hybrid bermudagrass cover as well as a reduction in sod tensile strength after applications of indaziflam at 0.03 kg ai ha 1 The authors also noted a decrease in p redicted bermudagrass cover at 10 WAT with applications of indaziflam (30% cover versus 100% for the nontreated check). Jones et al. (2013) reported an 89% reduction

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42 in bermudagrass root lengt h density from indaziflam (35 g ai ha 1 ) in a sandy soil with no organic matter during a greenho use study with mini rhizotrons. Based on the observed emergence pattern of TSG seedlings in this research, as well as the prediction model provided by Teuton et al. (2005), turfgrass managers should make PRE herbicide applications by the middle or end of March in order to optimize their results. An additional approach could be to follow up any TSG and/or turfgrass elimination with a POST herbicide application to target the newly emerged, and likely more susceptible, see dlings. Future research should identify selective POST herbicides that, although not highly efficacious against established TSG plants, may have increased efficacy against TSG seedlings. Amicarbazone has proven to be an effective option in combination with other POST herbicides (Cross et al. 2016). Teuton et al. (2004b) demonstrated >90% TSG control at the two to eight leaf stage in the greenhouse with compounds such as asulam and trifloxysulfuron, but was unable to achieve those levels of control in the f ield on more mature TSG plants. Also, future research should focus on additional PRE herbicides as well as the rate of reestablishment of the desired turf species into these eradicated areas to determine the most effective solution for TSG elimination foll owed by turfgrass reestablishment.

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43 Figure 3 1 Tropical signalgrass seedling emergence over t ime in the nontreated control and areas treated with S metolachlor and indaziflam. Weeks after treatment (WAT). Error bars correspond to standard error of the mean. 0 100 200 300 400 500 600 700 800 900 1 2 4 7 10 14 # seedlings m 2 WAT Control S-metolachlor Indaziflam

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44 CHAPTER 4 CONCLUSIONS AND RECOMMENDATIONS FOR FUTURE RESEARCH The banning of MSMA has greatly reduced the tools available to turfgrass managers for controlling TSG and other troublesome grass weeds in Florida. MSMA provided excellent control thr ough a unique mode of action and at an affordable price. Concerns over groundwater infiltration of highly toxic inorganic byproducts of MSMA ultimately led to its removal from the turfgrass market in Florida. Because herbicides are lost due to resistance d evelopment as well as environmental fate concerns, this research examined an integrated approach for controlling TSG in bermudagrass turf. Verticutting, a cultural practice used extensively in turfgrass management, provided a small amount of additional TSG control by itself through 12 WAA. The process of stolon fragmentation leads to reduced carbohydrate reserves from which axillary buds are able to draw on. Verticutting would be a useful component of an IWM strategy if it is already being done by turfgrass managers as it would provide additional control without any additional cost. Future research should be aimed at identifying additional cultural practices that are already implemented by managers, such as fertilization or irrigation regimes, which may be a dded to an IWM strategy in an attempt to either increase TSG control or accelerate turfgrass reestablishment into the newly vacated areas. PRE herbicides are an important component of TSG control. In a single study, cumulative TSG seedling counts reached u pwards of 800 seedlings m 2 over a period of 14 wk with a noticeable peak in emergence at 4 wk. Failure to eliminate as much of this emergence as possible would result in a turfgrass manager being forced to start over in their attempt to control TSG. Indaz iflam proved to be an effective option for controlling

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45 TSG PRE. In this same study, the indaziflam treatment resulted in >70% reduction in TSG seedling emergence compared to the nontreated check. In the IWM study, the addition of indaziflam to one of the P OST treatments resulted in nearly 50% TSG reduction at 52 WAA in comparison to the same treatment without the PRE herbicide. Future research should look at additional PRE herbicides such as oxadiazon and prodiamine to identify herbicides that are capable o f providing >90% control of TSG. Much of the soils in Florida are sandy, however, there are also areas with muck or clay soils that may be found on golf courses and sod farms. PRE, and potentially POST, herbicides may act very differently depending on the soil type that is present. Therefore, additional research is needed to determine the level of activity of indaziflam and other PRE herbicides on a variety of soils. Future research identifying methods of expediting turfgrass reestablishment should include plant growth regulators, such as trinexapac ethyl, that encourage lateral growth which could potentially speed this process up. Several POST herbicide combinations were identified in this research as having excellent (>90%) TSG control. The link between al l of these treatments was the PSII inhibiting compound amicarbazone, which demonstrated synergistic activity with both HPPD and ALS inhibitors. This ability to enhance other products could make amicarbazone a very useful component of IWM strategies. Futur e research should examine lower rates of amicarbazone with HPPD and ALS inhibiting compounds to identify effective treatments aimed at TSG (and other key weed species) control. These lower use rates would be more economically and environmentally preferabl e as well as provide less injury to the turfgrass.

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46 Of utmost importance, regardless of the herbicide applied, is the POST application timing in relation to projected TSG seedling emergence. If managed incorrectly, POST herbicide applications could ultimate ly become an extra cost with limited to no returns in TSG control. TSG POST eradication without proper PRE herbicide applications could allow for a flush of seedling emergence that would establish in the newly vacated areas before the turfgrass is able to do so. This could be further exacerbated by POST herbicide applications that also injure the bermudagrass turf, subsequently hampering its ability to reestablish into these areas. Therefore, burndown applications of glyphosate should always be accompanied by a PRE herbicide to combat potential TSG seedling emergence which could result in reestablishment.

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47 LIST OF REFERENCES Abendroth JA, Martin AR, Roeth FW (2006) Plant response to combinations of mesotrione and photosystem II inhibitors. Weed Technol 20:2 67 274 Anonymous (2012) Barricade 65WG herbicide product label. Greensboro, NC: Syngenta Crop Protection, LLC. 36 p Anonymous (2014) Xonerate herbicide product label. Cary, NC: Arysta LifeScience North America, LLC. 30 p Armel GR, Rardon PL, McComrick MC, Ferry NM (2007) Differential response to several carotenoid biosynthesis inhibitors in mixtures with atrazine. Weed Technol 21:947 953 Atwood D, Paisley Jones C (2017) Pesticides industry sales and usage 2008 2012 estimates. Washington, DC: U.S. Envi ronmental Protection Agency Barrett S (1983) Crop mimicry in weeds. Economic Botany 37:255 282 Bergkvist G, Ringselle B, Magnuski E, Mangerud K, Brandsaeter L (2017) Control of Elymus repens by rhizome fragmentation and repeated mowing in a newly establi shed white clover sward. Weed Res 57:172 181 Brosnan JT, Breeden GK (2013) Herbicide resistance in turfgrass: an emerging problem? Outlooks on Pest Management DOI: 10.1564/v24_aug_05 Bro snan JT, Breeden GK (2014) Use of MSMA for weed management in turf. The University of Tennessee Agricultural Extension Service W243 Brosnan JT, Breeden GK, McCullough PE, Henry GM (2012) PRE and POST control of annual bluegrass ( Poa annua ) with indaziflam. Weed Technol 26:48 53 Brosnan JT, Breeden GK, Thoms AW, Sorochan JC (2014) Effects of preemergence herbicides on the establishment rate and tensile strength of hybrid bermudagrass sod. Weed Technol 28:206 212 Brosnan J, Steckel L, Armel G (2009) W222 EPA decision on the use of organic arsenical herbicides for weed mana gement. The University of Tennessee Agricultural Extension Service Buhler DD (2002) Challenges and opportunities for integrated weed management. Weed Sci 50:273 280 Busey P, Johnston DL (2006 ) Impact of cultural factors on weed populations in St. Augusti ne turf. Weed Sci 54:961 967

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48 Chauhan BS, Johnson DE (2008) Germination ecology of goosegrass ( Eleusine indica ): an important grass weed in rainfed rice. Weed Sci 56:699 706 Chin HF (1979) Weed seed a potential source of danger. Pages 115 119 in Kwee LT ed. Proceedings of the plant protection seminar. Kuala Lumpur, Malaysia: Malaysian Plant Protection Society Colby SR (1967) Calculating synergistic and antagonistic responses of herbicide combinations. Weeds 15:20 22 Cole DJ (1994) Detoxification and activation of agrochemicals in plants. Pestic. Sci. 42:209 222 Cross RB, McCarty LB, Estes AG (2016) Postemergence tropical signalgrass ( Urochloa subquadripara ) control with nonorganic arsenical herbicides. Weed Technol 30:815 821 Cutulle MA, McElroy JS, Millwood RW, Sorochan JC, Stewart Jr. CN (2009) Selection of bioassay method influences detection of annual bluegrass resistance to mitotic inhibiting herbicides. Crop Sci 49:1088 1095 Danyal S, Bhowmik PC, Anderson RL, Shrestha A (2008) Revisiting the p erspective and progress of integrated weed management. Weed Sci 56:161 167 Dayan FW, Trindale MLB, Velini ED (2009) Amicarbazone, a new photosystem II inhibitor. Weed Technol 57:579 583 Fishel FM, Coats GE (1993) Effect of commonly used turfgrass herbici des on bermudagrass ( Cynodon dactylon ) root growth. Weed Sci 41:641 647 Integrated Weed Management. Weed Technol 27:1 11 Henry GM, Brosnan JT, Breeden GK, Cooper T, Beck LL, and Str aw CM (2012) Indaziflam programs for weed control in overseeded bermudagrass turf. HortTechnology 22:774 777 Hugie JA, Bollero GA, Tranel PJ, Riechers DE (2008) Defining the rate requirements for synergism between mesotrione and atrazine in redroot pigwee d ( Amaranthus retroflexus ). Weed Sci 56:265 270 Johnson BJ (1979) Vertical mowing and herbicide treatments on thatch in bermudagrass ( Cynodon dactylon ) turf. Weed Sci 27:14 17

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49 Jones PA, Brosnan JT, Kopsell DA, Breeden GK (2013) Soil type and rooting dept h affect hybrid bermudagrass injury with preemergence herbicides. Crop Sci 53:660 665 Mahoney DJ, Gannon TW, Jeffries MD, Polizzotto ML (2015) Arsenic distribution and speciation in a managed turfgrass system following monosodium methylarsenate applicatio n. Crop Sci 55:2877 2885 Matteson AR, Gannon TW, Jeffries MD, Haines S, Lewis DF, Polizzotto ML (2014) Arsenic retention in foliage and soil after monosodium methyl arsenate (MSMA) application to turfgrass. Journal of Environmental Quality 43:379 388 McC arty B, Estes A (2014) Tropical signalgrass control. Golf Course Manag 82:80 85 McCarty LB, Miller LC, Colvin DL (1991) Bermudagrass ( Cynodon spp.) cultivar response to diclofop, MSMA, and metribuzin. Weed Technol 5:27 32 McCullough PE, Johnston CR, Reed TV, Yu J (2015) Indaziflam enhances buckhorn plantain (Plantago lanceolate) control from postemergence herbicides. Weed Technol 29:147 153 Murphy TR, Colvin DL, Dickens R, Everest JW, Hall D, McCarty LB (1992) Weeds of southern turfgrasses. Florida Coope rative Extension Service Robertson JM, Taylor JS, Harker KN, Pocock RN, Yeung EC (1989) Apical dominance in rhizomes of quackgrass ( Elytrigia repens ): inhibitory effect of scale leaves. Weed Sci 37:680 687 Ryan GF (1970) Resistance of common groundsel to simazine and atrazine. Weed Sci 18:614 616 Shaner DL (2014) Herbicide Handbook. 10 th ed. Lawrence, KS: Weed Science Society of America Pp. 234 235, 245 246, 405 408, 443 444, 463 464 Soltani N, Dille JA, Burke IC, Everman WJ, VanGessel MJ, Davis VM, Sikkema PH (2016) Potential corn yield losses from weeds in North America. Weed Techol 30:979 984 Soltani N, Dille JA, Burke IC, Everman WJ, VanGessel MJ, Davis VM, Sikkema PH (2017) Perspectives on potential soybean yield losses from weeds in North Ameri ca. Weed Technol 31:148 154 Speedy A (2002) Brachiaria subquadripara (Tan.) Hitchc., Grassland Species. Food and Agricultural Organization. http://www.fao.org/ag/agp/agpc/doc/gbase /data/pf000194.htm Accessed: 03/21/2016

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50 Swanton CJ, Mahoney KJ, Chandler K, Gulden RH (2008) Integrated weed management: knowledge based weed management systems. Weed Sci 56:168 172 Taiz L, Zeiger E, Moller IM, Murphy A, eds (2015) Plant Physiology and Development. 6 th edn. Massachusetts: Sinauer Associates, Inc Teuton TC, Brecke BJ, Unruh JB, Macdonald GE, Miller GL, Ducar JT (2004a) Factors affecting seed germination of tropical signalgrass ( Urochloa subquadripara ). Weed Sci 52:376 381 Teuton TC, Mai n CL, Mueller TC (2005) Prediction modeling for tropical signalgrass ( Urochloa subquadripara ) emergence in Florida. Appl Turf Sci. DOI: 10.1094/ATS 2005 0425 01 BR Teuton TC, Unruh JB, Brecke BJ, MacDonald GE, Miller GL, Ducar JT (2004 b ) Tropical signalgr ass ( Urochloa subquadripara ) control with preemergence and postemergence applied herbicides. Weed Technol 18:419 425 Unruh JB, Partridge Telenko DE, Brecke BJ, Leon R (2009) ENH1132 Tropical signalgrass biology and management in turf. The University of Florida Environmental Horticulture Department [USDA] United States Department of Agriculture (2002) Urochloa subquadripara Natural Resources Conservation Service Plant Profile. http://plants.usda.gov/cgi_bin/topics.cg Vengris J (1962) The effect of rhizome length and depth at planting on the mechanical and chemical control of quackgrass. Weeds 10:71 74

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51 BIOGRAPHICAL SKETCH David Gregory Pearsa ul was born in Vero Beach, FL in 1982. He graduated from the University of Central Florida with a Bachelor of Science degree in criminal justice in 2004. After earning a second Bachelor of Science degree in biology, he began working at Syngenta Crop Protec tion where his passion for plant physiology began and, more importantly, he met his wife Samantha.