Two endemic late-season grape rot diseases, ripe rot and sour rot continue to pose a significant challenge to grape growers in Virginia and similar climates. Managing these diseases in the vineyard is challenging due to concerns over the increased risk of resistance against traditional single-site mode of action pesticides. This study evaluated the efficacy of different biopesticides and novel chemical options with a series of field trials across four vineyards in northern Virginia in 2021 and 2022. Among the tested products, a potassium-based nutrient formulation resulted in consistent ripe rot suppression in two of the three trials, showing around 46% and 83% less mean disease severity at the AHS AREC trial in 2022 and 2021, respectively, and 63% and 19% less mean disease incidence at the South Loudoun trial in 2021 and AHS AREC trial in 2022. For sour rot, zeta-cypermethrin with cyprodinil + fludioxonil or hydrogen peroxide + peroxyacetic acid resulted in effective control in two of the five trials, with a high of 71% and 73% less mean disease incidence compared to the untreated check in the Frederick trial in 2021 and 2022, respectively. In general, treatments with spinosad were less effective and resulted in lower disease control compared to those with zeta-cypermethrin and its fungicides/antimicrobials counterpart. The results suggest that integrating these biopesticides and novel chemicals into a spray program could offer a more sustainable solution for sour rot and ripe rot disease management without compromising the level of disease control. / Master of Science in Life Sciences / Ripe rot and sour rot are two diseases of concern for growers in Virginia and many other grape-growing regions due to significant losses in fresh cluster yield and the risk of contamination during the winemaking process. Traditional pesticides are often used; however, the breakdown of the effectiveness of these pesticides after continuous application, i.e., pesticide resistance, has become a critical issue. The other issues include, but are not limited to, adverse effects on human health, nature, and winemaking. On the other hand, biopesticides derived from microorganisms, their products, plant extracts, etc., are considered relatively safer and more sustainable options. This study evaluated the efficacy of biopesticides and novel chemicals against ripe rot and sour rot through field trials over two years at four locations in northern Virginia. Although no clear-cut winners were among the tested treatments, one of the tested chemicals (a potassium-based plant nutrient formulation) provided consistent disease suppression (up to 83%) against ripe rot. In the case of sour rot, several treatments, including a biologically derived insecticide, produced comparable disease suppression (up to 73%) compared to the current standard application. These safer options can be included in the spray schedule without compromising the level of disease control, meanwhile aiding the sustainability of the spray program in the long run.

Ripe rot of grape is caused by two fungal species complexes: <i>Colletotrichum gloeosporioides</i> and <i>C. acutatum</i>, both of which cause disease on a variety of crops, such as strawberry and apple. To investigate effect of cultivar and cluster developmental stage on the development of ripe rot, controlled environment and field studies were conducted during 2013-2014. We have identified that a certain level of infection could can take place on most cultivars tested from bloom to the near harvest. In most of the cases, significant cultivar and cluster developmental stage interaction effects were observed (<i>P</i> < 0.05) for the development of disease symptoms in both studies. In general, susceptible cultivar (Cabernet Franc, Cabernet sauvignon, and Chardonnay) demonstrated fluctuations of disease susceptibility among cluster development stages, while resistant cultivars (Merlot) showed consistently low level of the disease throughout the season. To investigate the effect of eleven ten modes of action for control of <i>C. gloeosporioides</i> and <i>C. acutatum</i>, two methods, alamarBlue® assay and inoculation on fungicide-treated detached fruits, were used. Protective fungicides (mancozeb, captan, and copper) as well as some of newer formulations such as azoxystrobin and tetraconazole were identified as excellent products against ripe rot of grape. Four additional materials were identified as good potential candidates to investigate further. The information gained from these studies will help growers to determine the critical period for ripe rot management and chemicals to be applied for management. With better control of cluster rot pathogens, Virginia growers can experience an increase in yield and wine quality. / Master of Science in Life Sciences

Colletotrichum acutatum and C. gloeosporioides are fungal plant pathogens that have a global distribution, extensive host range, and convoluted taxonomy. Both species can cause grape ripe rot and are considered endemic to Virginia US. In 2012, C. acutatum and C. gloeosporioides were reclassified into species complexes that consist of 31 and 22 accepted species, respectively. The objectives of this study were to: 1) survey Virginia vineyards for grape ripe rot, and morphologically and phylogenetically identify isolates to the species within the complexes, 2) conduct an in vitro fungicide assay to screen fifteen commercial fungicides and combinations of two fungicides for efficacy to control isolates from seven Colletotrichum species from Virginia vineyards, 3) sequence gene fragments from three subunits of the SDH enzyme in the fungicide-screened isolates to observe potential resistance mutations, 4) investigate the susceptibility of three grapevine tissues to Colletotrichum species, 5) observe potential infection structures before and after the application of fungicides, 6) evaluate the efficacy of commercial fungicide controls of grape ripe rot in the field, and determine the most advantageous timing of applications. In my studies, I identified six Colletotrichum species: C. aenigma, C. conoides, C. fioriniae, C. gloeosporioides, C. kahawae, and C. nymphaeae. I also found two additional groups; an isolate similar to C. limetticola and C. melonis and a group of isolates that are similar to C. alienum, C. fructicola, and C. nupharicola. I also identified captan, and mancozeb as two potential active ingredients for control of grape ripe rot isolates from Virginia via the in vitro fungicide assay. Additionally, I found that combinations of two active ingredients could increase the efficacy of benzovindiflupyr, copper, and polyoxin-D. C. fioriniae germination and production of melanized appressoria was documented on leaves. I observed appressorium formation with isolates of two C. fructicola-like genotypes and C. nymphaeae, as well as secondary conidiation with isolates of C. aenigma, C. fructicola-like genotype 3, and C. nymphaeae on blooms. And finally, benzovindiflupyr, cyprodinil + fludioxonil pre-mix, and potassium phosphite + tebuconazole were identified as candidates for chemical control for grape ripe rot in the field. / PHD / Colletotrichum acutatum and C. gloeosporioides are two fungal plant pathogens that are found on a wide range of crops around the globe. Both fungal species cause the disease grape ripe rot and have been found in Virginia (VA) USA since the late 1800s. Originally, grape ripe rot was considered a minor disease in VA; however, based on communications with local VA vineyard managers, grape ripe rot was found to cause up to 30% direct crop loss. Further indirect economic loss occurs during wine production due to the production of unpalatable, tobacco-like, off flavors from the infected grapes. Sensory studies found this wine flavor change occurred with as little as 3% of the total crushed grapes being infected. Grape ripe rot appears as a sunburn-like, tan injury on the surface of white-fruited grape berries. As the disease progresses, the dark injury expands across the surface of the berry and rings of salmon-colored spore masses form. On red-fruited grapes, the formation of spore masses is usually the first observable sign. Over time, the infected berries will shrivel down to a soft, pustule-covered raisin. Both C. acutatum and C. gloeosporioides cause the same grape ripe rot symptoms on fruit and overlap in fungal appearance. In addition, investigations of these pathogens using molecular techniques have revealed that each consists of a number of genetically distinct groups that are difficult to distinguish by appearance. Therefore, in 2012, C. acutatum and C. gloeosporioides were reclassified into 31 and 22 newly accepted species, respectively, using molecular techniques. The objectives of this study were to: 1) survey VA vineyards for grape ripe rot, and visually and molecularly identify isolates to the species within the new complexes, 2) conduct a laboratory fungicide assay to screen fifteen commercial fungicides and combinations of two fungicides for control of isolates from VA vineyards, 3) sequence gene fragments from three subunits of the SDH enzyme in the fungicide-screened isolates to observe potential resistance mutations, 4) investigate the susceptibility of three grapevine tissues to Colletotrichum species, 5) observe potential infection structures before and after the application of fungicides, 6) evaluate the efficacy of commercial fungicide controls of grape ripe rot in the field, and determine the most advantageous timing of applications. In my studies, I identified six Colletotrichum species: C. aenigma, C. conoides, C. fioriniae, C. gloeosporioides¸ C. kahawae, and C. nymphaeae. I also found two additional groups; an isolate similar to C. limetticola and C. melonis and a group of isolates that are similar to C. alienum, C. fructicola, and C. nupharicola. Our lab also identified four active ingredients as potential controls of grape ripe rot in the laboratory fungicide assay; captan, mancozeb, tetraconazole and thiophanate-methyl. Additionally, combinations of two compounds can increase the effectiveness of benzovindiflupyr, copper, and polyoxin-D. of C. fructicola-like isolates, and C. nymphaeae formed infection structures on blooms. of C. aenigma, C. fructicola-like genotype 3, and C. nymphaeae formed spores on blooms without producing symptoms. C. fioriniae spores germinated and produced infection structures on leaves without producing symptoms.