Genetic analysis of isolates of Botrytis cinerea sensitive and resistant to benzimidazole and dicarboximide fungicides

A total of 56 isolates of B. cinerea collected from ornamental crops from c ommercial greenhouses were examined by random amplified polymorphic DNA (RA PD) fingerprint analyses. Isolates were examined as two independent sets of 35 and 36 isolates, with 15 isolates common to both sets. The isolates had four phenotypes: 17 were sensitive to two commonly used fungicides, thioph anate-methyl (a benzimidazole) and vinclozolin (a dicarboximide) ((SSV)-S-T ); 18 were resistant to both fungicides ((RRV)-R-T); 16 were resistant to t hiophanate-methyl but sensitive to vinclozolin ((RSV)-S-T); and 5 were sens itive to thiophanate-methyl but resistant to vinclozolin ((SR)V)-R-T. Relat ionships among the isolates were determined by cluster analyses of mean cha racter differences using the unweighted pair group method using arithmetic average and cladograms were constructed. Isolates were clustered primarily by fungicide-sensitivity phenotype. In one set of greenhouse isolates, 6 of 10 (SSV)-S-T isolates clustered together with a bootstrap confidence value of 91%. In the other fingerprint set of greenhouse isolates, 9 of 11 (SSV) -S-T isolates clustered together and had a bootstrap confidence value of 98 %. Isolates resistant to thiophanate-methyl, vinclozolin, or both fungicide s usually were not clustered with other isolates or were clustered with iso lates of the same phenotype. To further elucidate these relationships, vari ant isolates resistant to one or both fungicides were produced on fungicide -amended agar medium from 14 (SSV)-S-T greenhouse isolates. These 14 (SSV)- S-T parent isolates, 57 resistant variant isolates, and 11 resistant greenh ouse isolates were analyzed as three independent RAPD fingerprint sets. Var iants selected from eight (SSV)-S-T parent isolates were resistant to both thiophanate-methyl and vinclozolin even though parent isolates were exposed to only one of the fungicides. Isolates resistant only to vinclozolin ((SR V)-R-T) had fingerprint patterns similar to and clustered with those of par ent isolates, while fingerprint patterns of isolates resistant to thiophana te-methyl (i.e., (RRV)-R-T or (RSV)-S-T), regardless of sensitivity to vinc lozolin, clustered differently from both those of STSV parent isolates and those of (SRV)-R-T isolates derived from the same parent. (RRV)-R-T and (RS V)-S-T variant isolates derived from the same fungicide-sensitive parents o nly clustered with other variants having the same phenotype.