The determination of resistance levels of the Botrytis cinerea isolates to some fungicides and their molecular characterization on grapes in Aegean region

Botrytis cinerea is the most destructive pathogen on grapes. The usage of very intensive fungicides to control the disease causes the risk of acquiring the pathogen resistance. In this study, Botrytis cinerea isolates were collected from the vineyards in the Aegean Region. Isolates were tested for their sensitivity to fungicides (pyrimethanil, iprodione, fenhexamid, cyprodinil+fludioxonil, boscalid) in vitro and in vivo conditions. Genetic differences between these isolates by molecular methods were also determined with specific primers. According to ED50 value of fungicides pyrimethanil and iprodione were found to low effective while fenhexamid and cyprodinil+fludioxonil were highly effective against mycelial growth of isolates. The efficacy on spore germination of B.cinerea isolates was determined at different doses of the fungicides. Cyprodinil+fludioxonil at the dose of 3 μg/ml inhibited spore germination of all B.cinerea isolates. It was observed that pyrimethanil did not inhibit mycelia growth of 50% of B.cinerea isolates and iprodione 70% in the dose range of 5-10 μg/ml. Both in vitro and in vivo conditions it was determined that cyprodinil+fludioxonil mixture was the most effective active ingredients to inhibit mycelial growth of isolates. All isolates had a point mutation (G143A). Therefore it was detected with allel-specific primers (BcAR-F; 5'-GGC AAA TGT CAC TGT GAG C-3', BcAR-R; 5'-ACC ATC TCC ATC CAC CAT ACC T-3'). Furthermore, to determine the genetic differences among B.cinerea isolates molecular analysis was carried out. In seven isolates of B.cinerea were obtained band 604 bp in gel electrophoresis of PCR products.