[Objective] The study aimed to investigate the genetic polymorphism of eighteen Lycium barbarum resources via nrDNA ITS sequencing. [Method] The genomic DNAs from Lycium barbarum leaves were isolated by modified CTAB ...[Objective] The study aimed to investigate the genetic polymorphism of eighteen 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 eighteen Lycium barbarum were firstly obtained in the present study. For all eighteen tested materials, the variation range of whole ITS region was 559-634 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 eighteen tested materials could be grouped into three classes. [Conclusion] Analysis of nrDNA ITS sequence may avail to identify the Lycium barbarum germplasm resources.展开更多
利用核糖体内转录间隔区(ITS)条形码序列,对枸杞杂交育种种内杂交种进行早期筛选研究。采用改进的十六烷基三甲基溴化铵(CTAB)法提取枸杞叶片DNA,利用合成的特异引物对其DNA中的nr DNA ITS区进行扩增、克隆,对目的片段进行测序分析。结...利用核糖体内转录间隔区(ITS)条形码序列,对枸杞杂交育种种内杂交种进行早期筛选研究。采用改进的十六烷基三甲基溴化铵(CTAB)法提取枸杞叶片DNA,利用合成的特异引物对其DNA中的nr DNA ITS区进行扩增、克隆,对目的片段进行测序分析。结果表明:以宁夏枸杞种内的品种宁杞1号、宁杞2号、白花枸杞作为父母本,选配杂交组合,基于ITS条形码序列对其种内杂交育种产生的杂交后代进行聚类分析,分析杂交后代与父母本的亲缘关系与差异,对其杂交后代进行早期筛选。由结果可知,基于ITS条形码序列可以作为早期筛选杂交育种后代的方法之一,对建立分子标记辅助育种技术、缩短育种周期具有重要意义。展开更多
文摘[Objective] The study aimed to investigate the genetic polymorphism of eighteen 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 eighteen Lycium barbarum were firstly obtained in the present study. For all eighteen tested materials, the variation range of whole ITS region was 559-634 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 eighteen tested materials could be grouped into three classes. [Conclusion] Analysis of nrDNA ITS sequence may avail to identify the Lycium barbarum germplasm resources.
文摘利用核糖体内转录间隔区(ITS)条形码序列,对枸杞杂交育种种内杂交种进行早期筛选研究。采用改进的十六烷基三甲基溴化铵(CTAB)法提取枸杞叶片DNA,利用合成的特异引物对其DNA中的nr DNA ITS区进行扩增、克隆,对目的片段进行测序分析。结果表明:以宁夏枸杞种内的品种宁杞1号、宁杞2号、白花枸杞作为父母本,选配杂交组合,基于ITS条形码序列对其种内杂交育种产生的杂交后代进行聚类分析,分析杂交后代与父母本的亲缘关系与差异,对其杂交后代进行早期筛选。由结果可知,基于ITS条形码序列可以作为早期筛选杂交育种后代的方法之一,对建立分子标记辅助育种技术、缩短育种周期具有重要意义。
