关键词: 灰葡萄孢, 细胞壁缺陷, T-DNA插入突变体, TAIL-PCR

Abstract: 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%．

Key words: Botrytis cinerea, cell wall defect, T-DNA insertion mutants, TAIL-PCR