一种钩藤属植物的分子鉴定

Molecular identification of one Uncaria plant

目的:以核糖体转录间隔区(rDNA ITS)序列为分子标记,对1种钩藤属植物归类到种提供分子证据。方法:改良CTAB法提取钩藤植物材料总DNA,通过引物对rDNA ITS区序列进行PCR扩增,经过克隆、测序后,与GenBank中已有的钩藤属植物rDNA ITS区序列进行比对,并应用Clustal X,BioEdit等软件计算分析,用PAUP软件构建系统发育树。结果:测序得到该钩藤植物的rDNA ITS区序列长度为719 bp,序列分析结果显示该钩藤植物与Genbank中已有的华钩藤Uncaria sinensisr DNA ITS区序列之间相似性达99.7%,并且在系统发育树中并排聚类成一支。结论:基于rDNA ITS区序列的测序分析和系统发育树构建的分子生物学方法,能够对该钩藤属植物进行准确的分子鉴定,为钩藤属中药材的种类鉴定和种间的分类地位提供分子生物学理论依据。

Objective：In order to identify a species of Uncaria,molecular phylogenetic analysis was carried out by using the rDNA ITS sequence as molecular marker.Method：Total DNA was extracted from the plant with modified CTAB method and thereby rDNA ITS regions were amplified with universally conserved primer.The rDNA ITS amplicon was characterized by cloning,sequencing,blasting in GenBank and phylogenetic analyses using PAUP by maximum parsimony（MP） criteria.Result：The rDNA ITS entire sequence of this species of Uncaria was 719 bp.The sequence is related to the U.sinensis available in GenBank and the similarity reaches 99.7%.Conclusion：Based on molecular biology methods of rDNA ITS region analysis,molecular identification is available in accurate classification on this species of Uncaria.