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


CONCLUSIONS
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.
Saito
Seiya
S
Laboratory of Fruit Genetic Engineering, The Institute of Enology and Viticulture, University of Yamanashi, Kofu, Yamanashi 400-0005, Japan.
Suzuki
Shunji
S
Takayanagi
Tsutomu
T
D001562:Benzimidazoles / Q000494:pharmacology; D020171:Botrytis / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D025141:Drug Resistance, Fungal; D005659:Fungicides, Industrial / Q000494:pharmacology*; D048448:Phenylcarbamates / Q000494:pharmacology; D010935:Plant Diseases / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D027843:Vitis / Q000254:growth & development* / Q000382:microbiology
D001562:Benzimidazoles; D005659:Fungicides, Industrial; D048448:Phenylcarbamates; C031000:benzimidazole