Efficacy and Feasibility of Application of SolviNix LC (Tobacco Mild Green Mosaic Tobamovirus) with A Wet-Blade Mower for Integration

with Mowing Brent A. Sellers, Jason A. Ferrell, and Raghavan Charudattan Abstract. A conventional practice for tropical soda apple (TSA; Solanum viarum) control in pastures is mowing, although it may be necessary to mow up to three times in order to kill TSA roots and prevent regrowth. Application of an herbicide to cut plant surfaces while mowing with a wet-blade mower might eliminate the need for repeated mowing. Tobacco mild green mosaic tobamovirus (TMGMV), a common naturally occurring plant virus has been formulated into a bioherbicide (SolviNix LC). SolviNix concentrations at 10 and 50 ug/ml were applied 9.4 and 18.8 L/ha; application rates on a per hectare basis were 0.24 g and 1.2 g for the 10 and 50 ug/ml concentrations applied at 9.4 L/ha and 0.47 and 2.3 g for the 10 and 50 ug/ml concentrations applied at 18.8 L/ha. Water was applied as the untreated control for both application volumes. The experiment was arranged in a randomized complete block design and was repeated at two locations in central and south Florida. Treatments were evaluated by counting plants in each 3 m by 60 m plot at 50 days after the wet-blade application. Control of TSA ranged from 57 to 59% with water alone at 9.4 and 18.8 L/ha, respectively. SolviNix did not result in greater control than water alone at any application rate, suggesting that either the application volume or an increased concentration may be required. Since excellent control with SolviNix has previously been demonstrated with a simulated wet-blade application, future testing should examine the use of increased application volumes or increased solution concentrations of the bioherbicide using wet-blade equipment. Nomenclature: TMGMV, Tobacco mild green mosaic tobamovirus; tropical soda apple, Solanum viarum Dunal, SOLVI.

Key words: wet-blade mower, invasive weed control, pasture weed control, tropical soda apple.

Tropical soda apple (Solanum viarum, TSA), a noxious weed (USDA-APHIS 2012), is a troublesome and costly invasive weed in Florida and the Southeast. In Florida alone, TSA infests about a million acres of open pastures, woodlands in cattle ranches, and natural areas (Mullahey et al. 1993; 1998; Langeland and Burks 2008). Dense TSA infestations hinder movement of cattle, workers, and management practices and force the stocking rates to be lowered. TSA seed is dispersed by cattle, water, and wildlife, and transported in contaminated manure, soil, hay, grass seeds, and sod; consequently, TSA has unrelenting potential to spread beyond its present distribution (Bryson and Byrd 2007; Cuda et al. 2002). Chemical herbicides, mowing, and biological control are used to manage TSA; a combination of methods and materials are needed, as no single approach is satisfactory under all conditions (Sellers et al. 2013). Research conducted at the University of Florida-IFAS has enabled the development of Tobacco mild green mosaic tobamovirus (TMGMV), a common, naturally occurring plant virus as a bioherbicide to control TSA (Charudattan and Hiebert 2007). A bioherbicide formulation (SolviNix LC) has been developed and successfully field-tested in Florida under an Experimental Use Permit. The EPA has reviewed a Section 3 registration package submitted by BioProdex, Inc., the SolviNix registrant and has accepted all, but have requested additional data on nontarget plant studies. A revised non-target plant studies data package will be submitted shortly for EPA review, and the registration outlook for SolviNix LC is considered good. The active ingredient in SolviNix, TMGMV, is a mechanically transmitted plant virus that normally infects plants through abrasion or wounding of the leaves. Because the virus has to be forced into the leaf tissues to initiate infection, application of formulated virus should be applied

using a high-pressure (> 80 psi) backpack sprayer or with an herbicide wiper that can simultaneously abrade and wipe the foliage with the bioherbicide solution. The high-pressure spot-treatment results in 80-100% TSA control, whereas application with the herbicide wiper yields 40-73% control (Charudattan et al. 2004). While the use of SolviNix for spot-treatment in natural areas such as woodlands is highly effective, practical, and provides a market niche for this bioherbicide, the ability to use it in open pastures will increase its overall acceptability as an alternative to chemical herbicides. A conventional practice for TSA control in pastures is mowing, although it may be necessary to mow up to three times in order to kill TSA roots and prevent regrowth. Application of an herbicide to cut plant surface while mowing with a wet-blade mower might eliminate the need for repeated mowing (Henson et al. 2003; Hixson et al. 2007; Wahlers et al. 1997). Previous work has demonstrated the feasibility and efficacy of application of a bacterial pathogen with a wet-blade mower to control TSA (DeValerio et al. 2011). Additionally, in a simulation of the wet-blade application, cutting TSA stems with hand-held pruning shears dipped in SolviNix prevents regrowth and kills the cut TSA stems (unpublished data). Given that mowing is an accepted TSA management practice, it is justified to attempt to integrate SolviNix LC with mowing. Our specific objectives are to test the feasibility of the concept with a commercial wet-blade mower, determine efficacy in terms of the level of TSA control achieved, active ingredient rate, and application volume. In practice, it would be easy to adopt this integrated method by producers. It would also improve user acceptance of SolviNix as a practical bioherbicide alternative to chemical herbicides. Materials and Methods

Experiments were established in TSA-infested pastures near Clewiston (26o2904.22 N, 80o5642.65 W) and Zolfo Springs (27o3129.87 N, 81o3458.25 W), Florida on 9 June, 2011 and 1 July, 2011, respectively. The experimental design was a 2 x 2 factorial treatment

arrangement in randomized complete block with four replications. Plot size was 4 by 66 m to ensure an adequate number of TSA plants. Size of TSA plants ranged from 15 to 51 cm at the time of treatment and the number of TSA plants in each plot was recorded prior to treatment. The bioherbicide treatment (TMGMV; SolviNix LC) was applied at 10 and 50 g/ml at 9.4 and 18.8 L/ha. Application rates on a per hectare basis were 0.24 g and 1.2 g for the 10 and 50 ug/ml concentrations, respectively, applied at 9.4 L/ha, and 0.47 and 2.3 g for the 10 and 50 ug/ml concentrations, respectively, applied at 18.8 L/ha. Water was applied in the untreated check for both application volumes. All treatments, including water, were applied using a 2 mwide wet-blade mower (Diamond Mowers, Sioux Falls, SD) set to clip at a height of 7.5 cm at a speed of 6.4 KPH. The efficacy of each treatment was evaluated by counting plants in each plot at 30 and 90 days after treatment (DAT). Count data were square root transformed prior to data analysis to satisfy the assumptions of ANOVA. Data were subjected to ANOVA using PROC GLM in SAS and were combined over locations as there was no location by treatment interaction and location was not significant in the model. Means were separated by Fishers Protected LSD (P = 0.05) when appropriate. Results and Discussion There were approximately 12 TSA plants in each plot prior to treatment at both locations (data not shown). Application volume did not affect the results. The number of TSA plants at 30 DAT ranged from 3.8 to 6.3 plants per plot and there were no differences between treatment

and the untreated control (Table 1). Similarly, there were no differences among treatments 90 DAT, and the number of TSA plants ranged from 0.4 to 2.1 plants per plot. Heavy rain and flooding in south Florida is the likely reason for decreased plant survival from 30 to 90 DAT as TSA plants do not typically survive under these conditions. Mowing TSA plants without herbicide application has been shown to provide some level of control. Mislevy et al. (1999) determined that a single mowing resulted in 47 and 60% TSA control 90 and 120 DAT, respectively. The height of the plants at the time of this mowing operation is unknown, but were reported to be mature with numerous fruits. The plants in this study had not begun fruit set, and this may be the reason for the higher level of control with mowing alone. The results of this research show that there was no difference in TSA control between the TMGMV treatments and the water-only checks. Previous research has shown that synthetic herbicide applications using the wet-blade system have been effective (Henson et al. 2003; Sellers and Mullahey 2008). However, application volumes utilized in the synthetic herbicide studies were applied at 25 L/ha. There is little information concerning the effect of lower application volumes using the wet-blade system; however, one could speculate that the application volumes used in this study were too low to obtain a biological effect using this system. This is especially true as cutting TSA stems with hand-held pruning shears dipped in TMGMV solution was shown to prevent regrowth under greenhouse conditions (unpublished data). Therefore, future research is needed to examine the effect of TMGMV on TSA control using the wet-blade system at applications volumes no less than 25 L/ha.

Acknowledgements. This research was funded by the IR-4 Biopesticide Research Program. The authors would also like to thank Diamond Mowers for use of the eco-blade mower for this research project.

References Bryson, C. T., J. Byrd, and R. G. Westbrooks. 1995. Tropical soda apple (Solanum viarum Dunal) in the United States. Fact Sheet. Bureau of Plant Industry, Mississippi Department of Agriculture and Commerce, Jackson, Mississippi, United States. Charudattan, R., M.S. Pettersen, and E. Hiebert. 2004. Use of tobacco mild green mosaic virus (TMGMV) mediated lethdal hypersensitive response (HR) as a novel method of weed control. U.S. Patent 997054. 18 p. Charudattan, R. and E. Hiebert. 2007. A plant virus as a bioherbicide for tropical soda apple, Solanum viarum. Outlooks Pest Manage. 18:167-171. Cuda, J.P., P.E. Parker, B.R. Coon, F.E. Vasquez, and J.M. Harrison. 2002. Evaluation of exotic Solanum spp. (Solanales: Solanaceae) in Florida as host plants for the leaf beetles Leptinotarsa defecta and L. Texana (Coleoptera: Chrysomelidae). Florida Entomol. 85:599-610. DeValerio, J.T., R. Charudattan, J.J. Mullahey, and P.D. Roberts. 2011. Application of a bacterial pathogen, Ralstonia solanacearum, with a wet-blade mower for biological control of tropical soda apple, Solanum viarum. Pest Technol. 5:102-107. Henson, S.E., W.A. Skroch, J.D. Burton, and A.D. Worsham. 2003. Herbicide efficacy using a wet-blade application system. Weed Technol. 17:320-324. Hixson, A.C., T.W. Gannon, and F.H. Yelverton. 2007. Efficacy of application placement equipment for tall fescue (Lolium arundinaceum) growth and seedhead suppression. Weed Technol. 21:801-806.

Langeland, K.A. and K.C. Burks. 1998. Identification and Biology of Non-Native Plants in Floridas Natural Areas. Institute of Food and Agricultural Sciences, University of Florida, Gainesville, Florida, United States. Mislevy, P., J. J. Mullahey, and F. G. Martin. 1997. Response of tropical soda apple (Solanum viarum) to triclopyr. Soil Crop Sci. Soc. Fla. Proc. 56:11-13. Mislevy, P., J.J. Mullahey, and F.G. Martin. 1999. Preherbicide mowing and herbicide rate on tropical soda apple (Solanum viarum) control. Weed Technol. 13:172-175. Mullahey, J.J., M. Nee, R.P. Wunderlin, and K.R. Delaney. 1993. Tropical soda apple (Solanum viarum): a new weed threat in subtropical regions. Weed Technol. 7:783-786. Sellers, B.A., J.A. Ferrell, J.J. Mullahey, and P. Hogue. 2013. Tropical soda apple: Biology, ecology, and management of a noxious weed in Florida (SS-AGR-77). Gainesville: University of Florida Institute of Food and Agricultural Sciences. Retrieved February 8, 2013, from http://edis.ifas.ufl.edu/uf097. Sellers, B.A. and J.J. Mullahey. 2008. Broadcast vs wet-blade herbicide applications for southern wax myrtle (Myrica cerifera) control. Weed Technol. 22:286-289. [USDA]-US Department of Agriculture Animal Plant Health Inspection Service. 2012. Federal noxious weed list. (http://www.aphis.usda.gov/plant_health/plant_pest_info/weeds/downloads/weedlist.pdf, 1 February 2012). Wahlers, R.L., J.D. Burton, E.P. Maness, and W.A. Skroch. 1997. A stem cut and blade delivery method of herbicide application for weed control. Weed Sci. 45:829-832.

Table 1. Response of tropical soda apple to Tobacco Mild Green Mosaic Virus (TMGMV) at 30 and 60 days after treatment (DAT). Application volume L/ha 9.4 ug/L 10 50 0 18.8 10 50 0 LSD (0.05) ----------------------------No./plot-----------------------12.6 16.8 12.4 11.6 12.8 11.8 NS 3.8 6.3 5.6 5.1 4.1 5.1 NS 0.4 1.4 1.0 0.5 2.1 0.4 NS TMGMV rate 0 DAT 30 DAT 60 DAT