不同地理居群新塔花的ITS2和psbA-trnH序列及聚类分析  

Cluster Analysis of Ziziphora bungeana from Different Geographical Populations Based on ITS2 and psbA-trnH Sequences

作　　者：苏艳 程波 何江 杨伟俊 SU Yan;CHENG Bo;HE Jiang;YANG Wei-jun(School of Food Science and Pharmacy,Xinjiang Agricultural University,Urumqi 830091,China;Key Laboratory of Xinjiang Uighur Medicine,Xinjiang Institute of Materia Medica,Urumqi 830004,China)

机构地区：[1]新疆农业大学食品科学与药学学院,乌鲁木齐830091 [2]新疆维吾尔自治区药物研究所新疆维吾尔药重点实验室,乌鲁木齐830004

出　　处：《中国实验方剂学杂志》2020年第16期175-182,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：新疆维吾尔自治区自然科学基金项目(2017D01B45)。

摘　　要：目的:分析18个不同地理居群新塔花的内转录间隔区(ITS)2和psbA-trnH序列,为其种质资源评价和基原药用植物遗传多样性分析提供参考。方法:试剂盒法提取新塔花基因组DNA,聚合酶链式反应(PCR)扩增ITS2和psbA-trnH间隔区序列,双向测序,拼接,基于Kimura两参数模型(K2P)构建邻接法(NJ)系统发育树。结果:不同地理居群新塔花ITS2和psbA-trnH序列均有种内差异。ITS2序列长度平均为236 bp,检测有9个单倍型,遗传距离为0~0. 022,不同地理居群新塔花聚为两支,XTH3,XTH6,XTH9等10个地理居群聚为一支,XTH4,XTH5,XTH10等8个地理居群聚为另一支。除XTH6的psbAtrnH序列存在6 bp缺失外,其他地理居群的psbA-trnH序列长度均为355 bp,检测有4个单倍型,遗传距离为0~0. 023,不同地理居群新塔花聚为两支,XTH1,XTH3,XTH4等12个地理居群聚为一支,XTH14,XTH17,XTH18聚为另一支。基于ITS2+psbA-trnH组合序列的系统发育(NJ)树显示,不同地理居群的新塔花可分为两支,XTH11,XTH12,XTH16等12个地理居群聚为一支,XTH14,XTH17,XTH18聚为另一支。结论:不同地理居群的新塔花地理位置接近或相似,相对遗传距离较小,亲缘关系较为接近,说明不同地理居群新塔花的亲缘关系及其遗传多样性与地理位置相关。Objective: To analyze the internal transcribed spacer(ITS)2 and psbA-trnH sequences of Ziziphora bungeana in 18 different geographic populations,in order to provide reference about evaluation of germplasm resources and analysis of genetic diversity of medicinal plants. Method: Genomic DNAs of the Z. bungeana were extracted by kit method. Polymerase chain reaction(PCR)was used to amplify ITS2 and psbA-trnH interstitial region sequences,bidirectional sequencing,splicing,and constructing Neighbor-joining(NJ)based on Kimura 2-parameter(K2 P)model. Result: All of sequences of ITS2 and psbA-trnH of Z.bungeana in different geographic populations showed intraspecific variations. The average ITS2 sequence length of Z. bungeana was 236 bp,9 haplotypes were detected,and the genetic distance was 0-0. 022. Z. bungeana of different geographical groups gathered into two branches,10 geographic populations,including XTH3,XTH6 and XTH9,were considered as one branch,while 8 geographic populations,including XTH4,XTH5 and XTH10,were the other branch. In addition to XTH6 that lacked 6 in bp psbA-trnH sequence,all of the other geographic populations had a 355 bp sequence of psbA-trnH,4 haplotypes were detected,and the genetic distance was 0-0. 023. 12 geographic populations,such as XTH1,XTH3,XTH4,gathered into one branch,while XTH14,XTH17 and XTH18 gathered into the other branch. NJ tree based on ITS2+psbA-trnH combination sequence showed that Z. bungeana of different geographical populations could be divided into two branches,with 12 geographical populations,like XTH11,XTH12,XTH16 as one branch,and XTH14,XTH17 and XTH18 as the other branch. Conclusion: Near or similar geographical locations of different geographical populations implies relatively short genetic distance and relatively close genetic relationship,which indicates that genetic relationship and genetic diversity of Z. bungeana in different geographical populations are related to geographical locations.

关 键 词：地理居群 新塔花 内转录间隔区2 PSBA-TRNH 聚类分析 遗传距离 

分 类 号：R284.2[医药卫生—中药学] R289[医药卫生—中医学]