基于nrDNA ITS序列的18份宁夏枸杞资源的遗传多样性

目的]利用nrDNA ITS序列,探讨18份宁夏枸杞资源的遗传多样性。方法]采用改进CTAB法提取枸杞叶片基因组DNA,利用合成的特异引物对其DNA中nrDNA ITS区进行扩增、克隆,对目的片段测序分析并对测序结果进行聚类分析。结果]通过测序首次获得了18种宁夏枸杞nrDNA ITS区碱基序列,整个ITS序列长度变异范围为559~634 bp,平均为612 bp,整个转录间隔区(ITS1+ITS2)对位排列后总长度为480 bp,包括194个变异位点,占40.4%;286个保守位点,占59.6%。聚类结果表明了18份宁夏枸杞的亲缘关系与差异,并将其分为3个大类。结论]基于nrDNA ITS区序列分析在研究枸杞种质遗传多样性方面具有一定的优越性。

Genetic Polymorphism of 18 Lycium barbarum Resources Based on nrDNA ITS Sequences

Objective] The study aimed to investigate the genetic polymorphism of 18 Lycium barbarum resources via nrDNA ITS sequencing.Method] The genomic DNAs from Lycium barbarum leaves were isolated by modified CTAB method for PCR amplification on the nrDNA ITS region using specifically synthesized primers;the amplified fragments were cloned and sequenced,then the sequencing results were clustered.Result] nrDNA ITS sequences of the tested 18 Lycium barbarum were firstly obtained in the present study.For whole 18 tested materials,the variation range of whole ITS region is 559-643 bp,with an average of 612 bp;alignment analyses showed that the whole length of internal transcribed spacer(ITS1 ITS2) was 480 bp,within which there are 194 variation sites,accounting for 40.4%,and 286 conserved sites,accounting for 59.6%.The cluster results showed that the 18 tested materials could be grouped into three classes.Conclusion] Analysis of nrDNA ITS sequence may avail to identify the Lycium barbarum germplasm resources.