堇菜属11种药用植物rDNAITS序列的克隆与分析  

Cloning and analysis of r DNA ITS sequences from 11 medicinal plants in Viola L.

作　　者：蒋明[1,2] 吴丹[3] 李嵘嵘[4] 管铭[1,2] 章燕如 

机构地区：[1]台州学院浙江省植物进化生态学与保护重点实验室,浙江椒江318000 [2]台州学院生命科学学院,浙江椒江318000 [3]杭州市中医院骨科,浙江杭州310007 [4]台州学院医药化工学院,浙江椒江318000

出　　处：《中草药》2015年第16期2454-2459,共6页Chinese Traditional and Herbal Drugs

基　　金：浙江省植物进化生态学与保护重点实验室植物进化生态学人才培育项目(2014)

摘　　要：目的通过测定和分析11种堇菜属药用植物的核糖体DNA内转录间隔区（r DNA ITS）序列,为该属药用植物的分子鉴定提供依据。方法利用PCR法克隆ITS序列,借助生物信息学工具进行序列分析、遗传距离估算和系统发育树构建。结果 11种堇菜属植物的ITS全长为612-638 bp,ITS1与ITS2的长度为251-265 bp和198-211 bp,5.8 S的长度十分保守,均为163 bp;ITS1、ITS2与5.8 S的信息位点数分别为50、23和3。11种堇菜属植物的遗传距离为0.025-0.137,紫花堇菜与七星莲的遗传距离最大,亲缘关系最远;紫花堇菜和鸡腿堇菜的遗传距离最小,亲缘关系最近。结论克隆到堇菜属11种药用植物的ITS序列,它们的信息位点丰富,可用于分子鉴定。Objective To provide molecular evidences for identification of medicinal plants in Viola L. by determining and analyzing the r DNA internal transcribed spacers（ITS） sequences of 11 species in Viola L. Methods ITS sequences were isolated using PCR amplification, and sequence analysis, evaluation of genetic distance as well as construction of phylogenetic tree were conducted by bioinformatics software. Results Full ITS sequences of 11 plants in Viola L. were varied from 612 to 638 bp in length. The lengths of ITS1 and ITS2 were 251—265 bp and 198—211 bp, respectively, while 5.8 S sequences were more conserved with identical length of 163 bp. Parsimony information sites of ITS1, ITS2, and 5.8 S were 50, 23, and 3, respectively. Genetic distance among 11 species in Viola L. varied from 0.025 to 0.137, and the greatest genetic distances were observed between V. grypoceras and V. diffusa var. brevibarbata, indicating their relative distinct relationship, and the smallest existed between V. diffusa and V. acuminata, revealing their closest relationship. Conclusion ITS sequences of 11 medicinal plants in Viola L. were obtained with a result of rich information sites, which provideds a foundation for molecular identification.

关 键 词：堇菜属 r DNA ITS 分子克隆 序列分析 

分 类 号：R282.12[医药卫生—中药学]