一株灰葡萄孢细胞壁完整性缺陷突变株的分子鉴定和表型分析  

Molecular identification and phenotypic analysis of a Botrytis cinerea mutant defective in cell wall integrity

作　　者：张为宏[1] 朱廷恒[1] 汪琨[1] 崔志峰[1] 

机构地区：[1]浙江工业大学生物与环境工程学院,浙江杭州310032

出　　处：《浙江农业学报》2012年第4期630-636,共7页Acta Agriculturae Zhejiangensis

基　　金：浙江省自然科学基金资助项目(Y307492)

摘　　要：为探明灰葡萄孢细胞壁相关基因功能,用荧光增白剂Calcofluor White(CFW)从灰葡萄孢T-DNA插入突变体库中筛选细胞壁完整性缺陷突变株,发现一株突变株D-59对CFW的敏感性与野生型相比提高了2.8倍。通过TAIL-PCR扩增突变株的T-DNA插入位点的侧翼序列并对其进行序列分析表明,T-DNA插入于突变株BC1G_00770.1基因的外显子部位。RT-PCR的结果显示,D-59的BC1G_00770.1基因不表达。突变株D-59的表型分析表明,突变株的菌丝稀疏,菌落呈土黄色,产孢量明显下降,孢子萌发异常,致病能力减弱,细胞壁的几丁质含量下降了48%。To study the cell wall related genes in Botrytis cinerea, T-DNA insertion mutants of Botrytis cinerea were screened by using Calcofluor White（CFW）, and a cell wall integrity defective mutant named D-59 was obtained. Compared with the wild-type strain, sensitivity of the D-59 mutant to CFW increased 2. 8 times. The flanking sequence of T-DNA insertion site was amplified by TAIL-PCR and the sequences were analyzed. It revealed that the T- DNA inserted in the extron part of a predicted protein coding gene （ BC1G_00770. 1 ） in B. cinerea genome. As ex- pected, no expression of BC1G_00770. 1 in the mutant was detected by RT-PCR experiment. It was found that D-59 showed sparse hyphae, khaki colonies, decreased spores, abnormal spore germination, decreased pathogenic ability, and the cell wall chitin content decreased by 48%.

关 键 词：灰葡萄孢 细胞壁缺陷 T-DNA插入突变体 TAIL-PCR 

分 类 号：S432.4[农业科学—植物病理学]