棉花学报2011，Vol.23Issue(1)：64-68,5.
棉花黄萎病菌插入突变体库的构建及致病相关基因 DVK1 的克隆与鉴定
Identification of the Pathogenicity-related Gene DVK1 in Verticillium dahliae
摘要
Abstract
In this study, we carried out T-DNA insertional mutagenesis to identify mutants, which are effective in pathogenicity.The highly virulent, defoliating strain was mutated through Agrobacterium tumefaciens-mediated transformation, and 5000 transformants were obtained.Pathogenicity test of randomly selected 1000 transformants found that five mutants lost their virulence on a susceptible cotton cultivar.A new mutant, d1, was unable to cause full disease on cotton.Analysis of the mutation using Thermal Asymmetric Interlaced PCR(TAIIL-PCR) confirmed an insertion into a gene DVK1.The full length of DVK1 genomic DNA and cDNA sequences were obtained using TAIL-PCR and RT-PCR methods.DVK1 has an open reading frame of 3325 bp interrupted by two introns with 52 bp and 36 bp, respectively, and putatively encodes a 1079 aa protein.The reduced pathogenic phenotype of d1 was fully complemented by reintroduction of the gene, indicating DVK1 is essential for pathogenicity in V.dahliae.关键词
T-DNA插入/突变体文库/基因克隆/棉花黄萎病/功能鉴定Key words
T-DNA insertional/ mutants library/ gene clone/ Verticillium dahliae/ functional identification分类
生物科学引用本文复制引用
高峰,彭姗,彭晓玲,李晖,李国英..棉花黄萎病菌插入突变体库的构建及致病相关基因 DVK1 的克隆与鉴定[J].棉花学报,2011,23(1):64-68,5.基金项目
农业部基础性工作专项(SB2007FY027) （SB2007FY027）
