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中国沙漠 ›› 2016, Vol. 36 ›› Issue (2): 374-382.DOI: 10.7522/j.issn.1000-694X.2016.00015

• 生物与土壤 • 上一篇    下一篇

荒漠植物红砂(Reaumuria soongarica)基因组微卫星富集文库的构建及特性

殷恒霞1,2, 石勇1,2, 张雯1, 燕霞1, 马小飞1   

  1. 1. 中国科学院寒区旱区环境与工程研究所 甘肃省寒区旱区逆境生理生态实验室, 甘肃 兰州 730000;<2r>2. 中国科学院大学, 北京 100049
  • 收稿日期:2016-01-09 修回日期:2016-02-24 出版日期:2016-03-20 发布日期:2016-03-20
  • 通讯作者: 马小飞,E-mail:maxiaofei@lzb.ac.cn
  • 作者简介:殷恒霞(1984-),女,青海西宁人,博士研究生,主要从事荒漠植物分子生态学研究。E-mail:hengxiayin@lzb.ac.cn
  • 基金资助:
    中国科学院"百人计划"项目(29Y127E71);国家自然科学基金项目(31370395,31500266)

Construction and Characterization of Enriched Microsatellite Library from Desert Plant Reaumuria soongarica

Yin Hengxia1,2, Shi Yong1,2, Zhang Wen1, Yan Xia1, Ma Xiaofei1   

  1. 1. Key Laboratory of Stress Physiology and Ecology in Cold and Arid Regions of Gansu Province, Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China;
    2. University of Chinese Academy of Sciences, Beijing 100049, China
  • Received:2016-01-09 Revised:2016-02-24 Online:2016-03-20 Published:2016-03-20

摘要: 采用磁珠富集法构建了红砂(Reaumuria soongarica)基因组微卫星富集文库,通过测序结果对文库的特性进行了分析。结果表明:在获得的13 922条微卫星序列中完美型占88%,非完美型10%,混合型2%。微卫星重复基元显示5种类型,以三碱基基元(GGT)n、(TGG)n、(ACC)n最多,占总数的60.7%;其次为二碱基基元,主要是 (AC)n、(GT)n、(CA)n,占38.6%;这5种微卫星重复基元的重复次数多集中在5~10。通过对随机挑选的100条微卫星序列设计引物,并以遗传距离和地理距离较远个体的基因组为模板进行扩增筛选,可有效扩增的引物为62对,并进一步测序筛选获得了12对多态性微卫星引物。磁珠富集法是开发红砂基因组微卫星序列的一种高效、可行的方法,构建的微卫星文库效率较高,富集量大,重复基元类型丰富,为红砂遗传进化和种质资源保护提供了可靠的数据支持和高效的分子遗传标记。

关键词: 红砂(Reaumuria soongarica), 微卫星, 磁珠, 富集文库

Abstract: The present study used the method of magnetic beads enrichment to build enriched microsatellite sequence library and analyzed the characteristics of library through sequencing results.The results showed that 13 922 microsatellite sequences were obtained with 88% perfect, 8% imperfect and 2% compound repeat motifs.Of these microsatellite loci, five type repeat primitives were presented, including prominent 60.7% three base primitive with (GGT)n, (TGG) n and (ACC)n, and 38.6% two base primitive with (AC)n, (GT)n and (CA)n. And the repeat time was mainly between 5 and 10, and declined as exceeding 30 times. Total 62 pairs of efficient PCR primers and 12 pairs of microsatellite primers have been screened by designing the primers with the 100 random microsatellite sequence and PCR screening with templates in populations with distant genetic and physical distance. Here, we revealed that the method of magnetic beads enrichment was high efficient and feasible for the development of Reaumuria soongarica genome microsatellite sequence. The present investigation would provide informative support and efficient genetic molecular marker for the genetic evolution and germplasm resources protection of Reaumuria soongarica.

Key words: Reaumuria soongarica, microsatellite, magnetic beads, enriched library

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