摘要
【目的】为准确、高效鉴别兔耳草属植物,保障药材质量,提高临床用药安全,对兔耳草属多种植物的遗传多样性进行研究。【方法】广泛收集青海省、云南省、四川省、西藏自治区等涉藏地区高海拔地区的9份短筒兔耳草和短穗兔耳草资源,采用PCR技术扩增其ITS2区段,并测序;从GenBank数据库中获取另外21份兔耳草属植物ITS2 DNA序列,使用MEGA5.5软件,寻找SNP位点,计算K-2p遗传距离,采用最大相似度法构建系统进化树。【结果】(1)短筒兔耳草和短穗兔耳草的ITS2序列中存在3个特异性变异位点。(2)短筒兔耳草、短穗兔耳草、狭苞兔耳草、紫叶兔耳草等兔耳草属12种植物的K-2p遗传距离为0.005~0.078,其中克什米尔兔耳草与亚中兔耳草的遗传距离最小,为0.005;裂叶兔耳草和倾卧兔耳草与匍匐兔耳草的遗传距离最大,均为0.078。(3)系统发育树分为两个部分,其中第一部分分为两个分支,全部的短筒兔耳草和短穗兔耳草亲缘关系很近,自展值为86,聚类于第一分支;来源于亚美尼亚的裂唇革叶兔耳草和来源于哈萨克斯坦的匍匐兔耳草自展值为93,聚类于第二分支。第二部分也分为两个分支,全部的紫叶兔耳草自展值为95,聚于一个分支;狭苞兔耳草、裂叶兔耳草、亚中兔耳草、倾卧兔耳草、小兔耳草、大筒兔耳草和克什米尔兔耳草自展值为90,聚于另一个分支。【结论】我国特有的短筒兔耳草和短穗兔耳草的亲缘关系较为接近,采用ITS2 DNA条形码可以有效地区分兔耳草属植物各物种。高海拔寒冷环境使兔耳草属物种形态变异较碱基变异明显,导致遗传分化较为强烈。
【Objective】In order to accurately and efficiently identify plants of the genus Lagotis,improve clinical drug safety,and ensure the quality of medicinal materials,the genetic diversity of various plants of the genus Lagotis was studied.【Method】Nine resources of L.brevituba and L.brachystachya were widely collected from high-altitude areas,such as Qinghai Province,Yunnan Province,Sichuan Province and Xizang Autonomous Region.The ITS2 segment was amplified by PCR and sequenced;ITS2 DNA sequences of the other 21 species of Lagotis were obtained from GenBank.MEGA5.5 software was used to find SNP loci,calculate K-2p genetic distance,and construct phylogenetic tree by maximum likehood method.【Result】(1)Three specific variation sites were found in ITS2 sequences of L.brevituba and L.brachystachya.(2)The K-2p genetic distance of 12 species of plants in the genus Lagotis,including L.brevituba,L.brachystachya,L.angustibracteata and L.praecox,ranged from 0.005 to 0.078.Among them,the genetic distance between L.cashmeriana and L.integrifolia was the smallest by 0.005;while the genetic distance between L.pharica,L.decumbens and L.stolonifera were the largest both by 0.078.(3)The phylogenetic tree was divided into two parts.The first part was divided into two branches.All of L.brevituba and L.brachystachya were clustered in the first branch,and the self-development value was 86,which proved that the two Lagotis species were closely related;L.korolkowii from Armenia and L.stolonifera from Kazakhstan were clustered in the second branch,with a self-development value of 93.The second part was also divided into two branches.All of L.praecox were clustered in one branch,and the selfdevelopment value was 95.L.angustibracteata,L.pharica,L.integrifolia,L.decumbens,L.minor,L.macrosiphon and L.cashmeriana were clustered another branch together,and the self-development value was 90.【Conclusion】The DNA barcoding of ITS2 could be used to effectively distinguish species of Rabdosia;The genetic relationship between L.brevituba and L.brachystachya endemic of China was close;high altitude and cold environment made the morphological variation of Lagotis species more obvious than base variation,resulting in strong genetic differentiation.
作者
李风琴
胡水清
杨梦婷
王宏璇
吴乐琦
吴波
LI Fengqin;HU Shuiqing;YANG Mengting;WANG Hongxuan;WU Leqi;WU Bo(College of Pharmacy,Jiangxi University of Chinese Medicine,Nanchang Jiangxi 330004,China;Research Center of Natural Resources of Chinese Medicinal Material and Ethnic Medicine,Jiangxi University of Chinese Medicine,Nanchang Jiangxi 330004,China;Attached Middle School of Jiangxi Normal University,Nanchang Jiangxi 330038,China)
出处
《南方林业科学》
2025年第3期6-10,17,共6页
South China Forestry Science
基金
“江西民族传统药现代科技与产业发展协同创新中心”资助项目(51316935)。
