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中国农学通报 ›› 2014, Vol. 30 ›› Issue (25): 128-135.doi: 10.11924/j.issn.1000-6850.2014-0740

所属专题: 生物技术

• 林学 园艺 园林 • 上一篇    下一篇

基于nrDNA-ITS序列的10种枸杞属植物亲缘关系研究

李小刚 李小刚 李捷 张晓娜 葛晶 何丽娟   

  • 收稿日期:2014-03-18 修回日期:2014-05-24 出版日期:2014-09-05 发布日期:2014-09-05
  • 基金资助:
    引进林业科学技术项目 “美国野生枸杞种质资源及培育利用技术引进” (948项目 2011-4-39)。

Study on 10 Species of Lycium L. Genetic Relationship Based on nrDNA-ITS Sequence

  • Received:2014-03-18 Revised:2014-05-24 Online:2014-09-05 Published:2014-09-05

摘要: 以10种枸杞属植物为材料,利用nrDNA-ITS序列,探讨其遗传多样性及亲缘关系。采用改进CTAB法提取枸杞叶片基因组DNA,利用合成的ITS4和ITS5为引物对其DNA中的nrDNA-ITS区进行扩增、对目的片段测序,并对测序结果进行聚类分析。测序结果表明,10种枸杞属植物ITS的长度为582~656 bp,保守位点(C)560个,变异位点(V)95个,其中简约信息位点(Pi)70个,单核苷酸变异位点(S) 25个;5.8S序列的长度均为154 bp,保守位点(C)136个,变异位点(V)18个,其中简约信息位点(Pi)12个,单核苷酸变异位点(S)6个。10条序列聚类图明显地分为2大类5个小组,‘0901’与‘蒙杞1号’处于同一分支,亲缘关系最近,‘宁杞3号’和‘宁杞7号’为同一分支,亲缘关系最近,‘宁杞5号’和青海诺木洪黑果枸杞单独为一分支,与其他几个枸杞品种的亲缘关系最远。10条枸杞属植物的ITS序列已上传至NCBI,登录号为KJ189761~KJ189770。本研究测序和分析了10种枸杞属植物的ITS序列,聚类结果表明了它们之间的亲缘关系与差异,为枸杞遗传多样性和系统发育研究奠定了基础。

关键词: 土地复垦, 土地复垦

Abstract: The nrDNA-ITS sequence was used to investigate the genetic diversity and genetic relationship of 10 Lycium L. species. After using improved method of CTAB to extracted genomic DNA from the leaves of Lycium L., the authors set synthetic ITS4 and ITS5 as primers to amplify the nrDNA-ITS region and sequence target fragment in those DNA at the same time, then analyzed the sequencing result through cluster analysis. The results were as follows: the ITS length in those 10 Lycium L. species was from 582 bp to 656 bp, and there were 560 conservative sites (C), 95 variable sites (V), which included 70 parsimony-informative sites (Pi) and 25 single nucleotide mutation sites (S); the length of 5.8S sequence were all 154 bp, and there were 136 conservative sites (C), 18 variable sites (V), which included 18 parsimony-informative sites (Pi) and 6 single nucleotide mutation sites (S). Those 10 sequences cluster figures of 10 Lycium L. species were classified into 2 categories and 5 groups apparently, the authors found that the‘0901’and‘Mengqi 1’were in the same branch, and‘Ningqi 3’and‘Ningqi 7’were also in the same branch, both two groups had the closest genetic relationship; meanwhile,‘Ningqi 5’and Lycium ruthenicum Murray from Nuomuhong Qinghai were in different branch, which had the furthest genetic relationship. The authors had already uploaded all 10 ITS sequences of Lycium L. species to the NCBI, and the NCBI accession numbers were from KJ189761 to KJ189770 accordingly. This study had sequenced and analyzed the ITS sequences of 10 Lycium L. species, and the cluster results indicated the genetic relationship and difference among them, which would lay theoretical foundation for future study in genetic diversity and system development of Lycium L.