猕猴桃溃疡病菌质粒的分离与功能分析
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国家自然科学基金项目(31401831);国家大学生创新训练项目(201610364018)


Functional analysis of natural plasmid from the pathogen of bacterial canker of kiwifruit, Pseudomonas syringae pv. actinidiae
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    摘要:

    分离猕猴桃溃疡病菌中的质粒,从质粒角度探索细菌的致病机制,分析其编码的主要基因、毒力因子。用试剂盒提取分离自安徽省岳西县猕猴桃溃疡病菌株JF8的质粒,对质粒DNA进行高通量测序及生物信息学分析。质粒基因组拼接显示菌株JF8质粒全长65 149 bp,GC含量56.29%,包含45个假定蛋白(hypothetical protein)和38个有预测功能的蛋白。生物信息学分析发现,质粒携带多达8个直接参与基因重组和转移的转运蛋白(mobile element protein),5个基因直接参与细菌Ⅳ型分泌系统、1个基因参与细菌Ⅲ型分泌系统。基因ParA编码的蛋白是一个ATP酶,对质粒的分配以及维持质粒稳定具有重要作用,编码转录激活蛋白的基因LuxR与细菌的群体感应机制相关,影响毒力因子的表达和分泌,基因UmuC是细菌中SOS应答系统的组分之一,控制细胞中聚合酶Ⅴ的量。

    Abstract:

    To explore the pathogenic mechanism of Pseudomonas syringae pv. actinidiae (Psa) from the perspective of plasmid,and to analyze the main genes and virulence factors they encode,we isolated high purity plasmid from the Psa strain JF8 (Yuexi,Anhui) by plasmid extraction kit,sequenced and analyzed it bioinformatically.The results showed that the plasmid was 50 724 bp with an average GC content of 56.29%,containing 83 protein-coding genes.Among the 83 genes,45 were predicted to encode hypothetical proteins and 38 were known functional genes.Bioinformatics analysis showed that the plasmid carried several genes associated with bacterial pathogenesis and disease resistance.The plasmid carries up to 8 mobile element proteins directly involved in gene recombination and metastasis.Five genes directly participate in the bacterial Ⅳ secretion system and one gene participates in the Ⅲ secretion system.The protein encoded by the gene ParA is an ATPase that plays an important role in the distribution of plasmids and maintaining plasmid stability.The gene LuxR,which encodes a transcriptional activator protein,is involved in the quorum sensing mechanism of bacteria and affects the expression and secretion of virulence factors.The UmuC gene is one of the components of the medium SOS response system,controlling the amount of polymerase Ⅴ in the cell.

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王月,董俊,吴群,汤小美,刘普.猕猴桃溃疡病菌质粒的分离与功能分析[J].华中农业大学学报,2018,37(3):39-45

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  • 收稿日期:2017-09-18
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  • 在线发布日期: 2018-05-03
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