The Role of Low Molecular Weight Fungal Metabolites in Grapevine Trunk Disease Pathogenesis: Eutypa Dieback and Esca

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 causative 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, Eutypa lata, Phaeoacremonium minimum and Phaeomoniella 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. IMPORTANCEUnderstanding the pathogenesis of grape trunk diseases (GTDs) is the key to the development of disease control and treatment. While fungal extracellular enzyme systems are typically cited relative to their fungal mechanisms in pathogenesis, non-enzymatic mechanisms have been less studied in this regard and the role of low molecular weight (LMW) fungal metabolites in GTD development is quite limited. In this article, we demonstrate that GTD-causative fungal pathogens Eutypa lata, Phaeoacremonium minimum and Phaeomoniella chlamydospore produce LMW phenolic metabolites under iron-restricted conditions. These metabolites undergo a series of redox reactions, with different fungi producing metabolites that preferentially either reduce iron, or generate hydrogen peroxide, under conditions simulating grapevine woody tissue. These conditions have the potential to promote generation of highly damaging hydroxyl radicals through a mechanism that appears to be similar to non-enzymatic chelator-mediated Fenton (CMF) chemistry which is involved in fungal degradation of wood by non-related fungal orders. This is the first report of CMF chemistry promoted by GTD-causative fungi under laboratory conditions and the research suggests an alternate pathway that may contribute to pathogenesis in GTDs, and a potential target for vine protection.