湖北后河国家级自然保护区短萼黄连的分子鉴定与分析  

Molecular identification and analysis of Coptischinensis var.brevisepala in Houhe National Nature Reserve,Hubei Province

作　　者：敖智广 韦乐华 杨蕾 李栋 李建斌 宋良科[1] 廖海[1] 茆灿泉[1] AO Zhiguang;WEI Lehua;YANG Lei;LI Dong;LI Jianbin;SONG Liangke;LIAO Hai;MAO Canquan(School of Life Science and Engineering,Southwest Jiaotong University,Chengdu 610031,China)

机构地区：[1]西南交通大学生命科学与工程学院,成都610031

出　　处：《生物学杂志》2021年第1期51-55,共5页Journal of Biology

基　　金：湖北五峰后河国家级自然保护区本底资源调查项目(2017G20001)。

摘　　要：使用DNA条形码分子鉴定技术,对湖北后河国家级自然保护区野生黄连物种进行分子鉴定,为保护区黄连的种质资源保护及潜在开发利用提供前期工作基础。共收集湖北后河、四川峨眉等不同产地的黄连植株及其近缘物种12份,对其进行DNA提取,PCR扩增和测序;对实验样本和GenBank下载的序列进行多序列比对、BLAST分析和NJ系统发育树构建。共获得黄连及其近缘物种ITS2序列82条、psbA-TrnH序列24条。不同种属黄连分别聚类。其中,黄连、短萼黄连种内平均K2P遗传距离分别为0.017和0.008,黄连属间为0.051。结合形态学鉴定和中药材DNA条形码技术,不同种属的黄连可以很好地区分。成功鉴定湖北后河国家级自然保护区野生黄连物种为短萼黄连(Coptischinensis var.brevisepala),其ITS2、psbA-TrnH序列长度分别为200 bp和258 bp。The DNA barcode to molecularly identify the wild Coptis species in Houhe National Nature Reserve,Hubei Province and provide a preliminary basis for the protection and potential development of Coptis in the protected area.The total DNA of 12 Coptischinensis and their close related species were extracted,ITS2 and psbA-TrnH PCR were amplified and sequenced.Homologous sequences were also downloaded and selected according to sequence alignment from GenBank database.All the sequences were aligned,and the neighbor-joining(NJ)phylogenetic trees were constructed.A total of 82 ITS2 sequences and 24 psbA-TrnH sequences were obtained.Different species of Coptis were clustered separately.The average K2P genetic distances for Coptischinensis,Coptischinensis var.brevisepala and Coptis spp.were 0.017,0.008 and 0.051,respectively.Combined morphological identification and the DNA barcode molecular identification technology,different Coptis species could be well distinguished and identified.The wild Coptis spp.in Houhe National Nature Reserve of Hubei was successfully identified as Coptischinensis var.brevisepala and their ITS2 and psbA-TrnH sequence lengths were 200 and 258 bp,respectively.

关 键 词：分子鉴定 黄连 ITS2 PSBA-TRNH 短萼黄连 

分 类 号：Q943.2[生物学—植物学] Q78