Department of Botany, University of Delhi, Delhi, 110 007, India.
School of Life Sciences, Jawaharlal Nehru University, Delhi, 110 067, India.
Sci Rep. 2017 Sep 6;7(1):10664. doi: 10.1038/s41598-017-09409-5.
Botrytis cinerea is a necrotrophic pathogen that infects many important crops. In an attempt to unravel some novel factors that govern pathogenicity in B. cinerea, Agrobacterium tumefaciens mediated transformation (ATMT) was deployed, and a number of tagged transformants were generated. Among these, a mutant, BCM-29 exhibited slower growth rate, reduced conidia size, conidiation and penetration. The mutant was also defective in secretion of oxalic acid (OA) and exhibited reduced activities of polygalacturonase (PG) and pectin methyl esterases (PME). TAIL-PCR followed by BLAST search identified the tagged gene as KLP-7 that encodes for kinesin. Targeted deletion of KLP-7 resulted in several folds decrease in virulence of mutants as compared to WT, while complementation of the gene helped in rescue of virulence traits. This is the first time when a unique kinesin KLP-7 that is mainly found in the phylum Pezizomycotina has been linked to virulence in B. cinerea.
灰葡萄孢是一种专性坏死型病原菌,可侵染许多重要作物。为了阐明一些控制灰葡萄孢致病性的新因子,我们采用根癌农杆菌介导的转化(ATMT)方法,并生成了许多标记转化体。其中,一个突变体 BCM-29 表现出较慢的生长速度、较小的分生孢子大小、产孢和穿透能力。该突变体还在分泌草酸(OA)方面存在缺陷,并且多聚半乳糖醛酸酶(PG)和果胶甲酯酶(PME)的活性降低。通过 TAIL-PCR 进行 BLAST 搜索后,鉴定出标记基因是编码驱动蛋白的 KLP-7。KLP-7 的靶向缺失导致突变体的毒力比 WT 降低了几个数量级,而该基因的互补有助于挽救毒力特征。这是首次将主要存在于 Pezizomycotina 门的独特驱动蛋白 KLP-7 与灰葡萄孢的致病性联系起来。