Identification of a novel point mutation in the β-tubulin gene of Botrytis cinerea and detection of benzimidazole resistance by a diagnostic PCR-RFLP assay

dc.contributor.author	Ziogas, BN	en
dc.contributor.author	Nikou, D	en
dc.contributor.author	Markoglou, AN	en
dc.contributor.author	Malandrakis, AA	en
dc.contributor.author	Vontas, J	en
dc.date.accessioned	2014-06-06T06:49:25Z
dc.date.available	2014-06-06T06:49:25Z
dc.date.issued	2009	en
dc.identifier.issn	09291873	en
dc.identifier.uri	http://dx.doi.org/10.1007/s10658-009-9462-y	en
dc.identifier.uri	http://62.217.125.90/xmlui/handle/123456789/4590
dc.subject	β-tubulin	en
dc.subject	Diethofencarb sensitivity	en
dc.subject	Molecular diagnostic assay	en
dc.subject	Monitoring resistance	en
dc.subject.other	amino acid	en
dc.subject.other	bioassay	en
dc.subject.other	detection method	en
dc.subject.other	fitness	en
dc.subject.other	fungicide	en
dc.subject.other	fungus	en
dc.subject.other	gene expression	en
dc.subject.other	genotype	en
dc.subject.other	identification method	en
dc.subject.other	laboratory method	en
dc.subject.other	molecular analysis	en
dc.subject.other	mutation	en
dc.subject.other	organic acid	en
dc.subject.other	organic compound	en
dc.subject.other	pesticide resistance	en
dc.subject.other	polymerase chain reaction	en
dc.subject.other	protein	en
dc.subject.other	Botryotinia fuckeliana	en
dc.title	Identification of a novel point mutation in the β-tubulin gene of Botrytis cinerea and detection of benzimidazole resistance by a diagnostic PCR-RFLP assay	en
heal.type	journalArticle	en
heal.identifier.primary	10.1007/s10658-009-9462-y	en
heal.publicationDate	2009	en
heal.abstract	The molecular basis of resistance to benzimidazole fungicides with laboratory and field mutant isolates of Botrytis cinerea was investigated. After chemical mutagenesis with N-methyl-N-nitrosogouanidine (NMNG) two different benzimidazole-resistant phenotypes were isolated on media containing carbendazim or a mixture of carbendazim and diethofencarb. The mutant isolates from the fungicide-mixture-containing medium were moderately resistant to carbendazim with wild-type tolerance to diethofencarb while mutant isolates from carbendazim-containing medium were highly resistant to carbendazim but sensitive to diethofencarb. The studied field isolates were highly resistant to benzimidazoles and sensitive to diethofencarb. Study of fitness characteristics of benzimidazole highly-resistant isolates showed that the resistance mutation(s) had no apparent effect on fitness-determining parameters. Contrary to this, the moderately benzimidazole-resistant strains, with no increased diethofencarb sensitivity, had a significant reduction in certain ecological fitness-determining characteristics. Analysis of the sequence of the β-tubulin gene revealed two amino acid replacements in the highly benzimidazole-resistant mutants compared to that of the wild-type parent strain. One was the glutamic acid (GAG) to alanine (GCG) change at position 198 (E198A), identified in both laboratory and field highly benzimidazole-resistant isolates, a mutation previously implicated in benzimidazole resistance. The second was a novel benzimidazole resistance mutation of glutamic acid (GAG) to glycine (GGG) substitution at the same position 198 (E198G), identified in a highly benzimidazole-resistant laboratory mutant strain. Molecular analysis of the moderately benzimidazole-resistant strains revealed no mutations at the β-tubulin gene. A novel diagnostic PCR-RFLP assay utilising a BsaI restriction site present in the benzimidazole-sensitive (E198) but absent in both resistant genotypes (E198G and E198A) was developed for the detection of both amino acid replacements at the β-tubulin gene. © KNPV 2009.	en
heal.journalName	European Journal of Plant Pathology	en
dc.identifier.issue	1	en
dc.identifier.volume	125	en
dc.identifier.doi	10.1007/s10658-009-9462-y	en
dc.identifier.spage	97	en
dc.identifier.epage	107	en