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天然橡胶作为重要的工业原料和战略资源,主要来源于巴西橡胶树.橡胶树炭疽病一直是导致其产量难以提高的重要叶部病害之一.效应蛋白作为病原菌侵染植物的重要致病因子,已成为植物与病原菌互作领域的研究热点.为了进一步阐明橡胶树炭疽菌的致病机理,本研究通过RT-PCR技术从橡胶树胶孢炭疽菌中克隆了一个候选效应蛋白基因,命名为CgE27,该基因编码一条121个氨基酸的多肽,分子质量为13.2kD.生物信息学分析表明,CgE27蛋白第1~第22位氨基酸是典型的信号肽序列,不含任何跨膜结构域,暗示该蛋白为分泌蛋白.依据同源重组原理,构建了 CgE27基因的敲除载体pCB1532-CgE27,并通过PEG介导转化法将其转入到胶孢炭疽菌原生质体细胞中进行同源交换,经过氯嘧磺隆抗性筛选和分子鉴定,成功获得敲除CgE27基因的胶孢炭疽菌敲除突变体株ΔCgE27,为进一步研究胶孢炭疽菌CgE27基因的功能提供了帮助.“,”Natural rubber, as an important industrial raw material and strategic resource, mainly comes from Brazil rubber tree. Anthracnose is one of the most important rubber tree leaf diseases that limits the yield of natural rubber increased. As an important pathogenic factor of pathogen infecting plants, the effectors become a research hotspot in the field of plant-pathogen interaction. In order to further elucidate the pathogenic mechanism of Colletotrichum gloeosporioides, a candidate gene from Colletotrichum gloeosporioides was cloned by RT-PCR and named as CgE27,which encoded a 121 amino acid polypeptide with a molecular mass of 13.2 kD. Bioinformatics analysis showed that the 1 ~22 amino acids of CgE27 protein was a typical signal peptide sequence with no transmembrane domain, suggesting that it was a secreted protein. According to the principle of homologous recombination, the knockout vector pCB1532-CgE27 of CgE27 gene was constructed, transformed into the hypha protoplast of Colletotrichum gloeosporioides by PEG-mediated transformation. After homologous exchange, screening and molecular identification, the C. gloeosporium knock-out mutant of CgE27 gene, ΔCgE27,was generated. ΔCgE27 would laid the foundation for further study on the function of CgE27.