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在农杆菌介导转化(AtMT)的稻瘟病菌突变体库中,筛选到一个菌丝生长增快,对大麦致病性增强的突变体B11。Southern杂交分析表明B11中T-DNA为单拷贝插入。通过TAIL-PCR克隆插入位点侧翼序列,序列测定与比对分析显示T-DNA位于假想基因MG01679的编码区内。利用PCR的方法,克隆基因的DNA和cDNA序列。该基因开放阅读框包括1个内含子和2个外显子,编码序列的长度为696 bp,编码231个氨基酸的多肽,编码产物属于ThiJ/PfpⅠ蛋白家族。因此,将该基因命名为MgThiJ1。MgThiJ1蛋白序列与尖胞镰刀菌假想蛋白FOXG_09029有57%的同源性,与禾谷镰刀菌假想蛋白FGSG_08979有54%的同源性。MgThiJ1基因可能为致病过程的负调控因子,其具体作用机制有待进一步研究。
In the mutant library of Magnaporthe grisea with Agrobacterium-mediated transformation (AtMT), a mutant B11 with rapid mycelium growth and increased pathogenicity to barley was screened. Southern hybridization analysis showed that T-DNA in B11 was inserted as a single copy. By TAIL-PCR cloning insertion site flanking sequences, sequencing and alignment analysis showed that the T-DNA is located in the coding region of the hypothetical gene MG01679. Using PCR methods, the DNA and cDNA sequences of the genes are cloned. The open reading frame of the gene contains an intron and two exons. The coding sequence is 696 bp in length and encodes a polypeptide of 231 amino acids. The encoded product belongs to the ThiJ / Pfp I protein family. Therefore, the gene was named MgThiJ1. The MgThiJ1 protein sequence has 57% homology with Fusarium solani hypothetical protein FOXG_09029 and 54% homology with Fusarium gramineus hypothetical protein FGSG_08979. MgThiJ1 gene may be a negative regulator of pathogenic process, its specific mechanism needs further study.