A New Flow Cytometry Technique to Identify Phaeomoniella chlamydospora Exopolysaccharides and Study Mechanisms of Esca Grapevine Foliar Symptoms

A flow cytometry technique that unequivocally identifies some of the toxic metabolites produced by Phaeomoniella chlamydospora, one of the main fungal pathogens causing esca disease of grapevine (Vitis vinifera), was developed. Antibodies raised against exopolysaccharides (EPS)— metabolites produced by Pa. chlamydospora that have been reported to be phytotoxic—were used as antigen to immunize rats. The specificity of these antibodies was assayed by flow cy-tometry against Pa. chlamydospora polysaccharides and against EPS with a different structure isolated from other phytopathogenic fungi, including Phaeoacremonium aleophilum and the Botryosphaeriaceae species Neofusicoccum luteum and N. parvum. Using this method, Pa. chla-mydospora polysaccharides were detected in the symptomatic leaves of esca-affected grape-vines, while healthy and asymptomatic leaves from both healthy and diseased vines did not pro-duce a binding reaction. This method potentially could be used to develop a simple kit to study the mechanisms underlying the development of esca foliar symptoms and to indirectly assess the presence of Pa. chlamydospora in grapevine material.