摘要
目的构建碎米桠Isodonrubescens及其与近缘种之间系统发育关系，准确鉴别碎米桠及其近缘种。方法提取碎米桠及其近缘种DNA，对核基因ITS片段和叶绿体基因psbA-trnH序列扩增和测序，计算物种种内、种间Kimura2-parameter（K2P）遗传距离。采用最近距离、相似性搜索、构建Neighbor-joining（NJ）系统聚类树3种方法进行鉴定分析。结果ITS和psbA-trnH分子系统树支持碎米桠5个居群样本聚为一单系分支，支持率为94％和68％；其中，碎米桠与毛叶香茶菜、香茶菜和内折香茶菜聚在一支，支持率为70％和51％；溪黄草和显脉香茶菜构成姐妹群分支，支持率为81％和63％。碎米桠及其近缘种种间ITS和psbA—trnH序列的K2P遗传距离为0．00708～0．07261和0．00806～0．02476，均远大于碎米桠种内的遗传距离0～0．00176和0～0．00268。结论碎米桠与毛叶香茶菜、香茶菜和内折香茶菜亲缘关系较近，溪黄草和显脉香茶菜亲缘关系最近。ITS和psbA-trnH序列可以准确鉴别碎米桠及其近缘种。
Objective To construct the phylogenetic relationships oflsodon rubescens and its relative species, and accurately identify L rubescens and its relative species. Methods Total genomic DNA was isolated from L rubescens and its relative species. Nuclear DNA internally transcibed spacer （ITS） and chloroplast gene psbA-trnIt sequences were amplified and sequenced. The Kimura 2-parameter （K2P） distances were calculated. Identification analyses were performed using Nearest Distance, BLAST1, and Neighbor-joining （NJ） methods. Results The NJ trees （ITS and psbA-trnH data） indicated that the five populations of I. rubescens formed a monophyletic clade [Bootstrap （BS） = 94% and 68%]. 1. rubescens and Ljaponicus, 1. amethystoides, I. inflexus formed one monophyletic clade （BS = 70% and 51%）, and I. serra was sister to 1. nervosus （BS = 81% and 63%）. The K2P genetic distances of ITS andpsbA-trnH data between L rubescens and its relative species were （0.007 08--0.072 61） and （0.008 06-0.024 76） which were far higher than intra-specific genetic distances of L rubescens （0-0.001 76） and （0-0.002 68）. Conclusion L rubescens is closely relative to I. japonicus, 1. amethystoides, and I. inflexus. The closest relative of L serra is I. nervosus. ITS and psbA-trnH are two efficient barcodes for the identification of I. rubescens and its relative species.
出处
《中草药》
CAS
CSCD
北大核心
2013年第20期2904-2909,共6页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金面上项目(31070171
31270236)
科技部国家星火计划项目(2012GA750004)
关键词
ITS
psbA—trnH
碎米桠
鉴定
亲缘关系
internally transcibed spacer
psbA-trnH
Isodon rubescens （Hemsl.） Hara
authentication
phylogenetic relationship
