title: Nested PCR-RFLP is a high-speed method to detect fungicide-resistant Botrytis cinerea at an early growth stage of grapes.
abstract: 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.
authors.affiliation: Laboratory of Fruit Genetic Engineering, The Institute of Enology and Viticulture, University of Yamanashi, Kofu, Yamanashi 400-0005, Japan.
authors.forename: Seiya
authors.initials: S
authors.lastname: Saito
authors.forename: Shunji
authors.initials: S
authors.lastname: Suzuki
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Takayanagi
mesh_terms: 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
chemical_list: D001562:Benzimidazoles; D005659:Fungicides, Industrial; D048448:Phenylcarbamates; C031000:benzimidazole