Nested PCR-RFLP is a high-speed method to detect fungicide-resistant Botrytis cinerea at an early growth stage of grapes

摘要

BACKGROUND:Grey mould caused by the fungus Botrytis cinerea Pers.ex Fr.is one of the major diseases in grapes.The use of fungicides is a simple strategy to protect grapes against B.cinerea disease.However,phenotypes exhibiting resistance to fungicides have been detected in B.cinerea populations.The variation of fungicide-resistant B.cinerea isolates renders B.cinerea disease control difficult in grapevine fields.RESULTS:The authors have developed a nested polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)method to detect fungicide-resistant B.cinerea isolates at an early growth stage of grapes in grapevine fields.The nested PCR-RFLP method was carried out to detect benzimidazole-,phenylcarbamate-and/or dicarboximide-resistant B.cinerea isolates from grape berries and leaves at Eichorn-Lorenz growth stage 25 to 29.This method successfully detected fungicide-resistant 6.cinerea isolates at an early growth stage of grapes.In addition,only 8 h was required from tissue sampling to phenotyping of fungicide resistance of the isolates.CONCLUSION:It is proposed that the early diagnosis of fungicide-resistant B.cinerea isolates would contribute to further improvement of integrated pest management against B.cinerea in grapevine fields,and that the nested PCR-RFLP method is a high-speed,sensitive and reliable tool for this purpose.