作 者：
许子欣;冉志芳;杨小彤;郝庆秀;余意;周洁;郭兰萍
单 位：
济南大学生物科学与技术学院;中国中医科学院中药资源中心无限极中草药品质研究联合实验室;无限极(中国)有限公司;山东中医药大学 药学院;济南大学 生物科学与技术学院
关键词：
巴戟天;居群;psbA-trnH;rDNA ITS;系统发育进化树;聚类分析
摘 要：
【目的】基于psbA-trnH和rDNA ITS序列进行不同地理居群巴戟天聚类分析,探讨不同地理居群巴戟天遗传变异与地理分布之间的相关性,为其道地性研究提供理论参考。【方法】以13个来自广东、广西和福建的不同地理居群巴戟天为研究对象,采用改良CTAB法提取其新鲜叶片DNA,以其为模板分别扩增psbA-trnH和rDNA ITS序列。采用ClustalX 1.81进行多重对位排列,应用MEGA 6.06中K2P(Kimura 2-parameter)模型计算遗传距离,并运用邻近法构建系统发育进化树。【结果】不同地理居群巴戟天的psbA-trnH序列长度为306 bp,GC含量为27.2%,保守率83.33%,变异率16.67%,简约信息率26.14%;不同地理居群巴戟天的rDNA ITS序列长度为571 bp,GC含量为64.1%,保守率91.42%,变异率8.58%,简约信息率14.19%。不同地理居群巴戟天的psbA-trnH序列遗传距离为0～0.128,其中广东省的5个样品间遗传距离整体较小;福建省的4个样品与广西和广东的样品间遗传距离整体较大。不同地理居群巴戟天的rDNA ITS序列遗传距离为0～0.102,其中福建省的永定2号与其他样品遗传距离均较大。基于psbA-trnH和rDNA ITS序列构建的系统发育进化树均将广东省的5个巴戟天样品聚为一支,但rDNA ITS序列构建的系统发育进化树还将广西区的4个巴戟天样品聚为一支,说明其可较好地区分广东省和广西区不同地理居群的巴戟天。【结论】巴戟天遗传结构与地理分布存在一定的相关性,深入研究巴戟天遗传变异与地理分布间的相关性时应以rDNA ITS序列为主、psbA-trnH序列为辅。
译 名：
Cluster analysis of Morinda officinalis How in different geographical populations based on psbA-trnH and rDNA ITS sequences
作 者：
XU Zi-xin;RAN Zhi-fang;YANG Xiao-tong;HAO Qing-xiu;YU Yi;ZHOU Jie;GUO Lan-ping;School of Biological Science and Technology,University of Jinan;School of Pharmaceutical Sciences,Shandong University of Traditional Chinese Medicine;The State Key Laboratory Breeding Base of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences;Infinitus(China) Company Ltd.;
关键词：
Morinda officinalis How;;populations;;psbA-trnH;;rDNA ITS;;phylogenetic tree;;cluster analysis
摘 要：
【Objective】Cluster analysis of Morinda officinalis How located in different geographical populations were analyzed based on PsbA-trnH and rDNA ITS sequences,and the relationship between genetic variation and geographical distribution was explored,which provided basic data for the researches of geoherb.【Method】A modified CTAB method was used to extract fresh leaf DNA from 13 different geographical populations of M.officinalis from Guangdong,Guangxi and Fujian.Specific primers were designed to amplify psbA-trnH and rDNA ITS sequences.The software of Clustal X 1.81 was used for multiple sequence alignment,and the pairwise distance was calculated based on K2P(Kimura2-parameter)model by MEGA 6.06 software,and the phylogenic tree was constructed by Neighbor-joining method.【Result】The psbA-trnH sequences of M.officinalis from different geographical populations were 306 bp,the GC content was27.2%,the conserved rate was 83.33%,the mutation rate was 16.67%,and the simple information rate was 26.14%.The rDNA ITS sequences of M.officinalis from different geographical populations were 571 bp,the GC content was 64.1%,the conserved rate was 91.42%,the mutation rate was 8.58%,and the simple information rate was 14.19%.The genetic distances of psbA-trnH sequences of M.officinalis from different geographical populations ranged from 0 to 0.128,among which the genetic distances among the five samples in Guangdong were small as a whole.And the genetic distances between four samples from Fujian and those from Guangxi and Guangdong were large.The genetic distances of rDNA ITS sequences of M.officinalis from different geographical populations ranged from 0 to 0.102.The genetic distances between Yongding 2 in Fujian and other samples were large.The phylogenetic tree based on psbA-trnH and rDNA ITS sequences could cluster five M.officinalis samples from Guangdong into one branch,but the phylogenetic tree based on rDNA ITS sequence could cluster four M.officinalis samples from Guangxi into one branch,which showed that the phylogenetic tree could better distinguish M.officinalis from different geographical populations in Guangdong and Guangxi.【Conclusion】There is certain correlation between genetic structure and geographical distribution of M.officinalis.The intensive study of the correlation between genetic variation and geographical distribution of M.officinalis should be based on ITS sequence and supplemented by psbA-trnH sequence.
