虾脊兰属植物DNA条形码的确立

Establishment of DNA barcode for the calanthe

目的:结合ITS2、psb A-trn H和matk序列对兰科虾脊兰属植物进行物种区分鉴定研究,并确立能准确鉴别虾脊兰属植物的序列或者序列组合。方法:采用试剂盒法对已收集的6种虾脊兰属植物进行总DNA的提取,通过扩增及测序,运用codencode V4.2软件对所得序列进行拼接与剪切,使用MEGA5.0对ITS2、psb A-trn H和matk序列3种序列分别进行比对分析,计算种内及种间Kimura 2-parameter(K2P)遗传距离,构建Neighbor-joining(NJ)系统聚类树。结果:实验共得到序列56条(包含Gen Bank序列17条),ITS2序列和mat K序列其种内最大遗传距离小于种间最小遗传距离,而该属psb Atrn H序列种内遗传距离范围和种间遗传距离范围有重合。结论:ITS2序列和mat K序列可以作为虾脊兰属部分植物的鉴定序列,而psb A-trn H序列不能用于虾脊兰属植物的鉴定。

Objective：Combined with the ITS2 and psbA-trnH and matK sequence to identify the medicinal plant-calanthe,and es-tablish genus sequences which can accurately identify the calanthe .Methods：Extracting total DNA from 6 species of calanthe , which had been collected by the kit method ,Then the sequences were amplified and sequenced ,Using codencodeV4.2 software to of splic and shear the sequences ,Comparing of all sequences of 6 species of the genus using MEGA5.0 software,The variable sites, the Kimura 2-Paramter（K2P）genetic distances and the Neighbor-joining（NJ）phylogenetic tree were computed by MEGA5.0.Re-sults：56 sequences（including 17 GenBank sequences）were obtained f.By analysis,ITS2 sequence and matK sequence of the in-traspecific genetic distance is less than the inter species genetic distance ,the intraspecific genetic distance and inter species genetic distance of psbA-trnH sequence was coincident .Conclusion：ITS2 sequence and matK sequence can be used as the identification sequence of the partial plants of the calanthe ,however psbA-trnH sequence could not be used for the identification of the calanthe .