title: RNA-mediated gene silencing in the phytopathogenic fungus Bipolaris oryzae.
abstract: The Ascomycetous fungus Bipolaris oryzae is the causal agent of brown leaf spot disease in rice and is a model for studying photomorphogenetic responses by near-UV radiation. Targeted gene disruption (knockout) for functional analysis of photomorphogenesis-related genes in B. oryzae can be achieved by homologous recombination with low efficiency. Here, the applicability of RNA silencing (knockdown) as a tool for targeting endogenous genes in B. oryzae is reported. A polyketide synthase gene (PKS1), involved in fungal DHN melanin biosynthesis pathways, was targeted by gene silencing as a marker. The silencing vector encoding hairpin RNA of the PKS1 fragment was constructed in a two-step PCR-based cloning, and introduced into the B. oryzae genomic DNA. Silencing of the PKS1 gene resulted in albino phenotypes and reduction of PKS1 mRNA expression. These results demonstrate the applicability of targeted gene silencing as a useful reverse-genetics approach in B. oryzae.
authors.affiliation: Faculty of Life and Environmental Science, Shimane University, Matsue, Shimane, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Moriwaki
authors.forename: Makoto
authors.initials: M
authors.lastname: Ueno
authors.forename: Sakae
authors.initials: S
authors.lastname: Arase
authors.forename: Junichi
authors.initials: J
authors.lastname: Kihara
mesh_terms: D001203:Ascomycota / Q000201:enzymology / Q000235:genetics*; D005656:Fungal Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D016681:Genome, Fungal; D010641:Phenotype; D048630:Polyketide Synthases / Q000037:antagonists & inhibitors* / Q000235:genetics; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D012333:RNA, Messenger; D048630:Polyketide Synthases