No abstract

Eutypa dieback and Esca complex are fungal diseases of grape that cause large economic losses in vineyards. These diseases require, or are enhanced by, fungal consortia growth which leads to the deterioration of the wood tissue in the grapevine trunk; however, pathogenesis and the underlying mechanisms involved in the woody tissue degradation are not understood. We examined the role that the consortia fungal metabolome may play the role in generating oxygen radicals that could potentially play a role in trunk decay and pathogenesis. Unique metabolites were isolated from the consortia fungi with some metabolites preferentially reducing iron whereas others were involved in redox cycling to generate hydrogen peroxide. Metabolite suites with different functions were produced when fungi were grown separately vs. when grown in consortia. Chelator-mediated Fenton (CMF) chemistry promoted by metabolites from these fungi allowed for the generation of highly reactive hydroxyl radicals. We hypothesize that this mechanism may be involved in pathogenicity in grapevine tissue as a causal mechanism associated with trunk wood deterioration/necrosis in these two diseases of grape.

Eutypa dieback and Esca are serious grapevine trunk diseases (GTDs) caused by fungal consortia causing large economic losses in vineyards. Depending on the disease the species involved include Eutypa lata, Phaeoacremonium minimum, and Phaeomoniella chlamydospora. There is a need to understand the complex pathogenesis mechanisms used by these fungi to develop treatments for the diseases they cause. Low molecular weight metabolites (LMW) are known to be involved in non-enzymatic oxygen radical generation in fungal degradation of wood by some Basidiomycota species, and as part of our work to explore the basis for fungal consortia pathogenesis, LMW metabolite involvement by the causal GTD fungi was explored. The GTD fungal pathogens examined, E. lata, P. minimum and P. chlamydospora, were found to produce low molecular weight iron binding metabolites that preferentially reduced iron or redox cycled to produce hydrogen peroxide. Uniquely, different LMW metabolites isolated from the GTD fungi promoted distinct chemistries that are important in a type of non-enzymatic catalysis known as chelator-mediated Fenton (CMF) reactions. CMF chemistry promoted by LMW metabolites from these fungi allowed for the generation of highly reactive hydroxyl radicals under conditions promoted by the fungi. We hypothesize that this new reported mechanism may help to explain the necrosis of woody grapevine tissue as a causal mechanism important in pathogenesis in these two grapevine trunk diseases.

Abamectin was tested for use with solid agar media in the laboratory to eliminate or kill the common mold mite Tyrophagus spp. in fungal cultures of Phaeomoniella chlamydospora (Pch) and Phaeoacremonium minimum (Pmin), two important grape pathogens involved in grapevine trunk disease. Abamectin concentrations tested were at or below the recommended dose for abamectin in greenhouse spray applications (≦625ug/mL) to control mites and determine if: a) fungal growth would be inhibited, and b) mites would be killed or their activity suppressed. Abamectin was added either to the media before autoclaving, or filter-sterilized and added after autoclaving, to test the effects of autoclaving on abamectin efficacy. Streptomycin (100µg/mL) was also added to a set of treatments to determine if this commonly-used antibiotic would impact abamectin efficacy against mites, or have an effect on fungal growth when in combination with abamectin. Filter-sterilized abamectin in the range of 62.5 - 312ug/mL, delivered to the media after it had been autoclaved, provided the most effective control of mites while also showing limited inhibition of fungal growth on solid agar media in the absence of streptomycin. The addition of filter-sterilized streptomycin had no significant effect on fungal growth for Pch, while for Pmin a small but significant reduction in growth with streptomycin occurred at abamectin concentrations above 62.5 ug/ml.