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灰葡萄孢产孢缺陷菌株的遗传分析

DOI: 10.7668/hbnxb.2011.03.018, PP. 86-89

Keywords: 灰葡萄孢,产孢量,TAIL-PCR,RT-PCR

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Abstract:

本研究从00余株灰葡萄孢ATMT转化子中筛选到产孢缺陷菌株BCG-183.利用CTAB法提取野生型和BCG-183的基因组DNA,Southern杂交证实T-DNA标记在突变菌株中为单拷贝插入;对该菌株表型进行分析,发现BCG-183菌株菌落呈白色,菌丝产量较野生型大;致病性较野生型强;利用血球计数板计算产孢量,表明BCG-183不产生分生孢子;胞壁降解酶活性比野生型强.利用TAIL-PCR技术确定T-DNA插入位点侧翼序列,生物信息学分析表明,T-DNA插入到灰葡萄孢基因组超级重叠群2(Supercontig2)中假定蛋白编码基因(BClG_0755)的3'末端.RT-PCR结果表明,BCJG_0755基因在突变菌株中没有表达.推测BC1G_0755可能与灰葡萄孢产孢相关.

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