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本研究通过筛选灰葡萄孢的ATMT突变体库,获得了1株不形成菌核,孢子产量明显减少,对番茄叶片的致病力明显增强的菌株BCt41。通过对该菌株进行PCR、Southern杂交鉴定,证明该菌株为遗传稳定的单拷贝T-DNA插入突变体。通过TAIL-PCR技术获得了T-DNA插入位点的侧翼序列,与灰葡萄孢数据库比对表明:T-DNA插入位点位于基因BC1G_02176.1的外显子2中。该基因编码区长1 206bp,含1个54bp内含子,编码383个氨基酸,基因功能未知。本研究结果为进一步研究该基因在灰葡萄孢分生孢子发育、菌核形成及致病过程中的作用奠定了基础。
In this study, we screened a library of ATMT mutant strains of Botrytis cinerea, and obtained one strain BCt41 which showed no significant growth of mycorrhizal spores and significantly reduced pathogenicity of tomato leaves. The strain was identified by PCR and Southern blot, which proved that the strain was a genetically stable single copy T-DNA insertion mutant. The flanking sequence of the T-DNA insertion site was obtained by TAIL-PCR. The comparison with the database of Botrytis cinerea showed that the T-DNA insertion site is located in exon 2 of BC1G_02176.1. The gene is 1 206 bp in length and contains a 54 bp intron, encoding 383 amino acids. The gene function is unknown. This study laid the foundation for further study on the role of this gene in conidial development, sclerotia formation and pathogenicity in Botrytis cinerea.