甘肃本地产当归、黄芪和大黄的rDNA ITS序列分析

Sequences Analysis of Ribosomal DNA ITS in Angelicae sinensis,Astragalus membranaceus and Rheum palmatum from Gansu Province

本研究采用改良CTAB法分别提取18份甘肃本地产当归、黄芪和大黄基因组DNA,并用PCR分别扩增其ITS1-5.8S-ITS2序列、直接测序并作序列同源性比对分析。双向测序分析结果表明,甘肃6个不同产地当归rDNA的ITS1、5.8S和ITS2序列一致,片段长度分别为215bp、162bp和223bp;供试的黄芪ITS1、5.8S和ITS2序列分别为228bp、164bp和210bp;大黄ITS1、5.8S和ITS2序列分别为160bp、159bp和164bp。供试材料的ITS1-5.8S-ITS2核苷酸序列已提交GenBank。本研究为提供甘肃当归、黄芪和大黄指纹图谱鉴别的分子标记、其道地性药材的分子鉴定和品质评价提供参考。

In this study,we have extracted 18 genomic DNAs of Angelicae sinensis,Astragalus membranaceus and Rheum palmatum from Gansu province by using modified CTAB method,and we also amplified the sequences of rDNA ITS1-5.8S-ITS2 of tested materials by PCR,respectively.Then we detected the rDNA ITS sequences of different species by direct sequencing and homological analyzing.The result of bidirection sequences alignment analysis showed that the sequences of Angelicae sinensis rDNA ITS1-5.8S-ITS2 were as the same with six different growing areas in Gansu,the length of Angelicae sinensis rDNA ITS1-5.8S-ITS2 was 215 bp,162 bp and 223 bp,respectively,and the length of Astragalus membranaceus rDNA ITS1-5.8S-ITS2 was 228 bp,164 bp and 210 bp respectively,the length of Rheum palmatum rDNA ITS1-5.8S-ITS2 was 215 bp,162 bp and 223 bp respectively.We have submitted the rDNA ITS sequences of all tested materials to GenBank,and all above research results would be a recommendable reference for the research on the molecular marker of fingerprint identification,the molecular identification of Chinese traditional medicines and evaluate the quality of Angelicae sinensis,Astragalus membranaceus and Rheum palmatum in Gansu province.