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植物研究 ›› 2016, Vol. 36 ›› Issue (2): 291-296.doi: 10.7525/j.issn.1673-5102.2016.02.019

• 论文 • 上一篇    下一篇

羌活(Notopterygium incisum)nrDNAITS和cpDNA rpl20-rps12序列分子进化特点的分析

杨路存1,2;刘何春1,3;周学丽4;徐文华1,2;周国英1,2*   

  1. 1.中国科学院西北高原生物研究所,810008;
    2.中国科学院藏药研究重点实验室,西宁 810008;
    3.中国科学院大学,北京 100049;
    4.青海省铁卜加草原改良试验站,西宁 810008
  • 出版日期:2016-03-15 发布日期:2016-05-20

Characteristic of Molecular Evolution of Notopterygium incisum Based on nrDNAITS and cpDNA rpl20-rps12 Sequence Analysis

YANG Lu-Cun1,2;LIU He-Chun1,3;ZHOU Xue-Li4;XU Wen-Hua1,2;ZHOU Guo-Ying1,2*   

  1. 1.Northwest Institute of Plateau Biology,Chinese Academy of Sciences,Xining 810008;
    2.Key Laboratory of Tibetan Medicine Research,Chinese Academy of Sciences,Xining 810008;
    3.University of Chinese Academy of Sciences,Beijing 100049;
    4.Tiebujia Grassland Improvement Experiment Station,Xining 810008
  • Online:2016-03-15 Published:2016-05-20

摘要: 应用PCR产物直接测序法分析了羌活居群间nrDNA(核糖体DNA)ITS序列和cpDNA(叶绿体DNA)rpl20-rps12的碱基差异,从而初步研究两套植物基因组的变异速率。采用改良的CTAB法从硅胶干燥的羌活叶片中提取总DNA,并对nrDNA ITS和cpDNA rpl20-rps12区域进行扩增、纯化、测序。nrDNAITS序列共有635 bp,有变异位点17处,变异位点百分率为2.68%,(G+C)含量为57.83%。cpDNA rpl20-rps12序列共有767 bp,有变异位点35处,变异位点百分率4.56%,(G+C)含量为33.06%。比较发现,羌活nrDNAITS区域较cpDNA rpl20-rps12序列保守,变异速率较慢。通过对ITS和rpl20-rps12序列单倍型(haplotype)进行分析发现,两者得出的结论一致,即现有分布范围经历了居群近期范围扩张。因此,羌活nrDNA ITS序列适合该种的谱系地理学研究。

关键词: 羌活, ITS序列, rpl20-rps12序列

Abstract: We analyzed the differences between the nrDNAITS and cpDNA rpl20-rps12 sequences in Notopterygium incisum by PCR direct sequencing. Total DNA was extracted from silica-dried leaves of N.incisum using modified CTAB method. With the extracted DNA as template, nrDNA ITS and cpDNA rpl20-rps12 regions were amplified, then purified and sequenced. The length of nrDNA ITS sequence of N.incisum was 635 bp, of which 17 were variable sites with a percentage of 2.68%, the(G+C) content was 57.83%. The length of cpDNA rpl20-rps12 sequence of N.incisum was 767 bp, of which 35 was variable site with a percentage of 4.56%, the(G+C) content was 33.06%. The nrDNAITS region of N.incisum was more conserved and evolved more slowly than the cpDNA rpl20-rps12sequence. The present distribution range of N.incisum experienced range expansion by the haplotype analysis of this species, which consisted with the conclusion resulting from cpDNA genome. Therefore, the nrDNA ITS sequence of rpl20-rps12 was fit to the phylogeographic study of this species.

Key words: Notopterygium incisum, ITSsequence, rpl20-rps12 sequence