[关键词]
[摘要]
目的 通过叶绿体基因psbA-trnH序列分析,探讨我国忽地笑种质资源的系统进化关系及分子鉴定方法。方法 分别提取15省(市)52个忽地笑居群的DNA,经PCR扩增叶绿体基因psbA-trnH序列及测序,并用Mega 5.0等软件对测序结果进行分析。结果 52条psbA-trnH序列长度为544~656 bp,GC含量为35.8%~37.0%,遗传距离为0.000 00~0.009 47;核苷酸变异(多态性)位点数共33个,其中简约信息位点9个,单一突变位点18个,插入/缺失片段6个;单倍型数量(H)10个,单倍型多态性水平(Hd)0.749,核苷酸多态性(π)0.002 63,收集的忽地笑资源具有较高的遗传多样性。最大简约法(maximum parsimony,MP)系统树中52个居群聚为4类,并且该聚类结果与其地理分布基本一致。结论 不同产地忽地笑居群的遗传变异较大,psbA-trnH序列可作为忽地笑种源分子鉴定的依据;我国忽地笑种质资源在进化上具有明显的地域性特征。
[Key word]
[Abstract]
Objective System evolution relationship and molecular identification method of the germplasm resources of Lycoris aurea from different regions was analyzed based on the sequence of psbA-trnH chloroplast gene. Methods DNA samples of 52 L. aurea populations were extracted from 15 provinces or cities in China. The psbA-trnH sequences of the populations were amplified by PCR, and the purified PCR products were sequenced and analyzed by Mega 5.0 software etc. Results The length of psbA-trnH sequences were 544-656 bp, and GC content of them was 35.8%-37.0%, and the genetic distances among the populations were 0-0.009 47. There were 33 variable (polymorphic) sites, including nine parsimony informative sites and 18 singleton variable sites and six insertion/deletion gaps. Ten haplotypes (H) were identified. Values of haplotype diversity (Hd) and nucleotide diversity (π) were 0.749 and 0.002 63, respectively. The genetic diversity of the populations of L. aurea were very high. In the maximum parsimony phylogenetic tree, 52 populations of L. aurea were clustered into four branches, which was almost consistent with their geographical distributions. Conclusion The genetic variation of L. aurea populations from different regions is significant and the psbA-trnH sequence could be used as a molecular evidence for identifying the germplasm resources of L. aurea from different regions. There is very obvious regional characteristics in evolution for germplasm resources of L. aurea in China.
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[基金项目]
国家自然科学基金资助项目(31470403);湖南省双一流学科建设经费资助