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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Phylogeny, histological observation, and in vitro fungicide screening and field trials of multiple Colletotrichum species, the causal agents of grape ripe rot

Oliver, Charlotte 31 January 2019 (has links)
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.

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