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水稻条斑病菌内源质粒分析 预览

Analysis of indigenous plasmids in Xanthomonas oryzae pv. oryzicola
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摘要 【目的】明确水稻条斑病菌(Xanthomonas oryzae pv. oryzicola,简称Xoc)内源质粒分布状况、类型及其含有的主要功能基因,为下一步开展该菌与质粒相关的毒力进化研究、抗逆机制分析及穿梭载体构建打下基础。【方法】采用数据库检索法和质粒分离鉴定法,调查水稻条斑病菌内源质粒的分布状况;以GenBank数据库中Xoc菌株基因组序列为研究对象,开展质粒信息检索,并对已发表的Xoc内源质粒序列进行系统进化分析;以Xoc中国菌株菌种库为研究对象,采用碱裂解法提取质粒,进行质粒酶切带型分析和主要功能基因鉴定。【结果】GenBank全基因组数据库中Xoc菌株内源质粒检出率为11.76%,系统进化分析结果表明这些质粒属于不同的进化分支。在257株Xoc中国菌株中,有29株菌株含有质粒,质粒检出率为11.28%,带有内源质粒的菌株均来自于华南稻区,且大部分来自广西。通过质粒酶切图谱比较,确认新发现的质粒属于9种新型Xoc内源质粒。按照已知Xoc内源质粒pXOCgx01上主要功能基因序列设计引物,PCR扩增结果表明新鉴定的Xoc质粒含有重金属抗性和DNA转移相关的基因,但在菌株间存在明显的多样性。【结论】部分Xoc菌株中含有一定数量的内源质粒,质粒类型和所含功能基因等具有一定的多样性,携带内源质粒的Xoc菌株有明显地域性。 【Objective】The aim was to fully understand the distribution,types and biological function of the indigenous plasmids of Xanthomonas oryzae pv. oryzicola(Xoc),and lay the foundation for the future study of virulence evolution analysis,mechanism of resistance and construction of shuttle vector.【Method】The distribution of endogenous plasmids in Xoc strains was investigated by database mining and plasmid isolation. With Xoc strain genome sequence in Gen- Bank as the research target,plasmids information searching was conducted,and phylogenetic analysis on published Xoc indigenous plasmids sequence was also carried out. With Xoc Chinese strains database as the research target,the plasmids were extracted by alkaline lysis,plasmidrestriction endonuclease pattern was analyzed and main functional genes were identified.【Result】The detection rate of Xoc strainindigenous plasmids in GenBank whole genome database was 11.76%. The phylogenetic analysis indicated that these plasmids were clustered into different phylogenetic branches. Among the 257 Xoc Chinese strains,29 contained plasmids with a detection rate of 11.28%. The strains containing indigenous plasmid all came from southern China,and most of them were from Guangxi. Nine novel indigenous plasmids of Xoc were confirmed by restriction endonuclease pattern. The primers were designed based on main functional gene sequence of the known indigenous plasmid pXOCgx01. The PCR amplification analysis indicated that the novel plasmids contained heavy metal tolerance and DNA transposition related genes,but there was diversity among the strains.【Conclusion】Partial Xoc strains contain indigenous plasmids,with diversities in plasmid types,functional genes(DNA transposition and heavy metal resistance),and the Xoc strains with indigenous plasmids have obvious geographical origins.
作者 曾晨 李康佳 尹朝群 牛祥娜 赵嘉城 朱平川 姜伟 何勇强 ZENG Chen;LI Kang-jia;YIN Chao-qun;NIU Xiang-na;ZHAO Jia-cheng;ZHU Ping-chuan;JIANGWei;HE Yong-qiang(State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources/College of Life Science and Technology,Guangxi University,Nanning 530004,China)
出处 《南方农业学报》 CAS CSCD 北大核心 2019年第8期1718-1727,共10页 Journal of Southern Agriculture
基金 国家重点研发计划项目(2018YFD02003021) 国家自然科学基金项目(31660505) 公益性行业(农业)科研专项(201303015).
关键词 水稻条斑病菌 内源质粒 调查 酶切图谱 抗逆基因 Xanthomonas oryzae pv. oryzicola indigenous plasmid general survey restriction pattern resistance gene
作者简介 曾晨(1993-),研究方向为植物病原细菌功能基因组学,E-mail:3137314963@qq.com;通讯作者:何勇强(1963-),博士,教授,主要从事植物病原细菌功能基因组学研究工作,E-mail:yqhe@gxu.edu.cn。
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