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 expected, 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%.%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％.
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