Secondary metabolite production by the fungal pathogen Eutypa lata:: Analysis of extracts from grapevine cultures and detection of those metabolites in planta

Eutypa dieback of grapevines is caused by the fungal pathogen Eutypa lata and reduces vineyard longevity worldwide. Early detection could reduce losses due to this disease, so our aim was to identify acetylenic phenol metabolites of E. lata that could prove suitable as chemical markers in an early diagnostic test for the pathogen. Accordingly, secondary metabolite production by 30 isolates of E. lata grown on media derived from canes of three grapevine cultivars was analysed using HPLC. Six metabolites, namely eutypinol, methyl eutypinol, eulatachromene, eutypine, 2-iso-propenyl-5-formylbenzofuran and eulatinol, were detected in culture filtrates. Most abundant were eutypinol and methyl eutypinol, produced by 97 and 83% of isolates, respectively. There was no apparent correlation between secondary metabolite production on media containing milled canes from the three cultivars of grapevine, and the field tolerance of these same cultivars to Eutypa dieback. When various other fungi commonly isolated from grapevine trunks in Australia were grown on milled cane, no secondary metabolites characteristic of E. lata were detected, suggesting such compounds are specific to E. lata. To examine the detection of secondary metabolites in planta, micropropagated grapevine plantlets were treated with purified or crude culture filtrates from nine isolates of E. lata grown on malt yeast broth. Various secondary metabolites were identified in,treated plantlets, however, no single compound was detected consistently. Eutypinol was detected in micropropagated grapevine plantlets inoculated with mycelium of E. lata, however, no metabolites were detected in the sap of vines which had been artificially inoculated with the pathogen.