黄芪属植物DNA条形码与聚类分析的研究

DNA barcoding sequences and cluster analysis of 10 species of Astragalus

目的通过分析吉林产10种黄芪属植物的ITS2、matk、rbcL、psbA条形码序列,为黄芪属植物的物种分类鉴定及其亲缘关系提供理论依据。方法共收集30份样品,以ITS2、matk、rbcL和psbA作为条形码序列,对黄芪属10种植物提取基因组DNA、PCR扩增并进行双向测序。所得序列结果经校正处理后,并对其进行聚类分析。结果ITS2、matk、rbcL和psbA4种片段,单独的每一种片段并不能将10种植物全部鉴别出来,只能鉴别出部分植物,即蒙古黄芪A.membranceus var.mongholicus、扁茎黄芪A.complanatus、糙叶黄芪A.scaberrimus、华黄芪A.chinensis、草木樨黄芪A.melilotoides、细叶黄芪A.angustissimus、兴安黄芪A.dahuricus、新巴黄芪A.hsinbaticus;由聚类分析结果可知,以锦鸡儿Caraganasinica为外类群,10种植物中,华黄芪为单独1支,细叶黄芪与扁茎黄芪聚为1支,糙叶黄芪、斜茎黄芪与新巴黄芪聚为1支,膜荚黄芪A.membranceu、蒙古黄芪与草木樨黄芪聚为1支,并且这3者与兴安黄芪关系较近。结论ITS2+rbcL+psbA组合可以将8种黄芪属植物鉴别出来。聚类分析结果显示10种黄芪属植物被划分为5支,该聚类分析结果与《中国植物志》中10种黄芪属植物的传统分类相一致。

Objective To provide theoretical basis for species classification and phylogenetic relationship of Astragalus by analyzing ITS2,matk,rbcL,psbA barcodes of 10 species of Astragalus grown in Jilin Province.Methods Based on 30 samples of 10 species of Astragalus,genomic DNA was extracted and amplified by PCR and bi-directionally sequenced with ITS2,matk,rbcL and psbA as barcode sequences.The sequence results were corrected and cluster analysis was carried out.Results Each of four fragments of ITS2,matk,rbcL and psbA could not identify all 10 plants,but only some plants,eg.A.membranaceus var.mongholicus,A.complanatus,A.scaberrimus,A.chinensis,A.melilotoides,A.angustissimus,A.dahuricus and A.hsinbaticus.From the results of cluster analysis,with Caragana sinica as the outer group,A.chinensis was a single branch in 10 species,A.angustissimus and A.complanatus were one branch,A.scaberrimus,A.laxmannii and A.hsinbaticus were one branch,A.membranceus,A.membranaceus var.mongholicus and A.melilotoides were one branch,and they were closely related to A.dahuricus.The result of cluster analysis was consistent with the traditional classification of 10 species of Astragalus L.in Flora of China.Conclusion The combination of ITS2+rbcL+psbA could identify eight species of Astragalus.Ten species of Astragalus grown in Jilin Province are divided into five branches by cluster analysis,which is consistent with the traditional classification of 10species of Astragalus in Flora of China.