基于松材线虫全基因组序列的SSR标记开发

许峻荣; 吴小芹; 刘云; 叶建仁

doi:10.3969/j.issn.1000-2006.2014.02.008

许峻荣,吴小芹,刘云,叶建仁

南京林业大学学报（自然科学版） ›› 2014, Vol. 38 ›› Issue (02) : 36-42.

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南京林业大学学报（自然科学版） ›› 2014, Vol. 38 ›› Issue (02) : 36-42. DOI: 10.3969/j.issn.1000-2006.2014.02.008

研究论文

基于松材线虫全基因组序列的SSR标记开发

许峻荣,吴小芹^*,刘云,叶建仁

作者信息 +

Development of simple sequence repeats base on pine wood nematode (Bursaphelenchus xylophilus)genome sequence

XU Junrong, WU Xiaoqin^*, LIU Yun, YE Jianren

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文章历史 +

摘要

为了研究松材线虫在我国的群体遗传关系及传播路径,获得更为稳定的松材线虫分子标记,使用MISA软件对松材线虫全基因组10 432条DNA片段进行搜索,共获得95个gSSR位点。其中,二核苷酸重复出现频率最高,占全部SSR位点的66.3%。依据所有gSSR位点共设计出36对引物,以1份松材线虫DNA pooling(包含我国46个不同地理来源的松材线虫虫株DNA)为模板进行PCR,产物由QIAxcel全自动凝胶电泳分析系统检测,获得17对可能具有多态性的gSSR引物。进一步对这17对引物的PCR产物进行单克隆试验并测序,BioEdit软件拼接比对结果表明,其中9对确实具有多态性。

Abstract

To develop more stable molecular marker, SSR marker base on pine wood nematode(Bursaphelenchus xylophilus)whole genome sequences was discussed for researching the relationship of population genetic and travel route accurately. Ninety-five SSR loci were searched from a total of 10 432 DNA fragments of B.xylophilus by MISA. In the gSSRs, the dinucleotide repeat motifs were the most abundant(66.3%). Thirty-six pairs of primers were designed, and verified with one peace of B.xylophilus DNA pooling sample(46 DNA samples of isolates from different geographic origin contained)for PCR. The products detected by QIAxcel automatic gel electrophoresis analysis system showed that 17 pairs of primers had potential polymorphism. Further, nine pairs of the primers showed polymorphism, which tested by monoclonal sequencing and spliced by BioEdit, and which provided foundation for studying population genetic of B.xylophilus.

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许峻荣,吴小芹,刘云,叶建仁. 基于松材线虫全基因组序列的SSR标记开发[J]. 南京林业大学学报（自然科学版）. 2014, 38(02): 36-42 https://doi.org/10.3969/j.issn.1000-2006.2014.02.008

XU Junrong, WU Xiaoqin, LIU Yun, YE Jianren. Development of simple sequence repeats base on pine wood nematode (Bursaphelenchus xylophilus)genome sequence[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2014, 38(02): 36-42 https://doi.org/10.3969/j.issn.1000-2006.2014.02.008

中图分类号： Q78 S763.16

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基金

收稿日期:2013-04-23 修回日期:2013-07-23
基金项目:国家自然科学基金项目(31270683); 江苏省高校自然科学研究重大项目(11KJA22002); 江苏高校优势学科建设工程资助项目(PAPD); 江苏省普通高校研究生科研创新计划项目
第一作者:许峻荣,博士。*通信作者:吴小芹,教授。E-mail: xqwu@njfu.edu.cn。
引文格式:许峻荣,吴小芹,刘云,等. 基于松材线虫全基因组序列的SSR标记开发[J]. 南京林业大学学报:自然科学