Open AccessRNA‐mediated gene silencing in the phytopathogenic fungus Bipolaris oryzae
Author(s) -
Moriwaki Akihiro,
Ueno Makoto,
Arase Sakae,
Kihara Junichi
Publication year - 2007
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2006.00606.x
Subject(s) - gene silencing , biology , gene , rna silencing , genetics , gene knockdown , rna interference , small hairpin rna , polyketide synthase , trans acting sirna , gene expression , rna , polyketide , biosynthesis
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 .