一种钩藤属植物的rDNA ITS序列分析

The phylogenetic study of one Genus Uncaria plant based on rDNA ITS

目的以核糖体转录间隔区(r DNA ITS)序列作为分子条形码,对一种常用中药材钩藤进行分子鉴定和遗传分析,并阐明其在钩藤属内种间的分子系统发育关系。方法通过改良十六烷基三甲基溴化铵(CTAB)法对2015年7月采集的钩藤进行总DNA提取,利用通用引物对rDNA ITS序列进行PCR扩增,经克隆、测序后,运用MEGA软件进行系统发育分析和构建系统发育树。结果测序得到该钩藤的rDNA ITS区序列长度为718 bp,序列分析结果显示其与Gen Bank数据库中已有的钩藤(Uncaria rhynchophylla)rDNA ITS区序列之间相似度达99.8%,并且在系统发育树中并排聚类成一支。结论基于ITS区序列测定分析的分子生物学方法和构建系统发育树的分子遗传学方法,可以对钩藤属药用植物进行分子鉴定和遗传分析,为钩藤属药用植物的种间分类地位以及种类鉴定提供准确的分子证据。

Objective To clarify the phylogenetic relationship of one species of Uncaria,molecular identification and genetic analysis were carried out by using the rDNA ITS sequence as molecular barcoding. Methods Molecular identi- fication and genetic analysis were carried out with the rDNA ITS sequence as molecular marker.Total DNA was ex- tracted with modified CTAB method and thereby rDNA ITS regions were ：amplified with universal primer, sequenced and phylogenetic analysed with MEGA 6.0 at July 2015, Results The entire rDNA ITS sequence of Uncaria was 718 bp.Se- quence analysis showed that the rDNA ITS sequence was closely related tO Uncaria rhynchophylla available in Gen- Bank and the similarity reached 99.8%. Conclusion Based on molecular biology methods of rDNA ITS region analysis, molecular identification is available in accurate classification on traditional Chinese medicinal material plants in Genus Uncaria and provide molecular evidence for taxonomy and identification of different species in Genus Uncaria.