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对农杆菌介导转化引起的致病性减弱突变菌株H800的菌落形态、孢子形态、菌丝体生长速率、孢子萌发率和致病力等生物学特性进行研究,并利用TAIL-PCR方法克隆T-DNA插入位点基因,结合生物信息学分析该基因功能与H800表型变化及致病性减弱的关系。结果表明,突变菌株H800的致病力低于野生型菌株CH008;生长速率为0.83 cm/d,亦显著低于CH008的1.13 cm/d;产孢量和分生孢子大小与CH008无明显差异;孢子萌发率为2.58%,显著低于野生型的95.98%;序列分析显示,T-DNA已插入St Cg800基因的启动子区域,基因结构分析表明,St Cg800基因编码区全长774 bp,无内含子,可编码257个氨基酸多肽的St Cg800蛋白。该蛋白是不稳定的水溶性蛋白,亚细胞定位于细胞质,且无信号肽,推测为生长因子类的酶,但功能尚未知。
The biological characteristics such as colony morphology, spore morphology, mycelium growth rate, spore germination rate and pathogenicity of the mutant H800 strain induced by Agrobacterium-mediated transformation were studied. The TAIL-PCR method was used to clone T DNA insertion site genes, combined with bioinformatics analysis of the gene function and H800 phenotypic changes and pathogenicity of the relationship. The results showed that the virulence of mutant strain H800 was lower than that of wild type strain CH008; the growth rate was 0.83 cm / d, which was also significantly lower than that of CH008 (1.13 cm / d); the sporulation and conidia size had no significant difference with CH008; The spore germination rate was 2.58%, which was significantly lower than 95.98% of the wild type. Sequence analysis showed that T-DNA was inserted into the promoter region of St Cg800 gene. The gene structure analysis showed that the coding region of St Cg800 gene was 774 bp in length, Intron, St Cg800 protein encoding a 257 amino acid polypeptide. The protein is an unstable water-soluble protein, subcellular localization in the cytoplasm, and no signal peptide, presumed to be growth factor enzymes, but the function is unknown.