title: Heterologous expression of the CYP51 gene of the obligate fungus Blumeria graminis in the necrotrophic fungus Botrytis cinerea.
abstract: As it is extremely difficult to make DNA transformation for the obligate fungus, Blumeria graminis f. sp. tritici (Bgt), we developed a heterologous expression system for characterization of a Bgt gene, CYP51, which encodes 14α-demethylase. The CYP51 gene from Bgt was transformed into the necrotrophic fungus, Botrytis cinerea. Reverse transcription polymerase chain reaction showed that the Bgt CYP51 was transcribed in B. cinerea. Green fluorescence was observed in the transformants of B. cinerea carrying the Bgt CYP51-GFP fusion cassette, suggesting that its translation was successful. Fungicide sensitivity tests revealed that B. cinerea transformed with Bgt CYP51 showed reduced sensitivity to a sterol demethylation inhibitor triadimefon, but not to a benzimidazole fungicide carbendazim. These results indicated that this heterologous expression system can be used for functional analysis of other Bgt genes.
authors: Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China.
authors: Lei-Yan
authors: LY
authors: Yan
authors: Yan-Feng
authors: YF
authors: Chen
authors: Qian-Qian
authors: QQ
authors: Yang
authors: Zhong-Hua
authors: ZH
authors: Ma
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology; D001203:Ascomycota / Q000201:enzymology* / Q000235:genetics*; D001562:Benzimidazoles / Q000494:pharmacology; D002219:Carbamates / Q000494:pharmacology; D003001:Cloning, Molecular; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D058886:Sterol 14-Demethylase / Q000235:genetics*; D014170:Transformation, Genetic; D014230:Triazoles / Q000494:pharmacology
chemical_list: D000935:Antifungal Agents; D001562:Benzimidazoles; D002219:Carbamates; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014230:Triazoles; D049452:Green Fluorescent Proteins; C032910:triadimefon; D058886:Sterol 14-Demethylase; C006698:carbendazim