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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: To provide molecular evidence for the classification of one genus Uncaria to be based on the sequence of ribosomal transcribed spacer (rDNA ITS). Methods: The total DNA of Uncaria tomentosa plant was extracted by modified CTAB method. The rDNA ITS region was amplified by PCR. After cloning and sequencing, the rDNA ITS sequence of GenBank was compared with that of GenBank. Application of Clustal X, BioEdit and other software calculation and analysis, the use of PAUP software to build phylogenetic tree. Results: The sequence of rDNA ITS region was 719 bp. The sequence analysis showed that the similarity of the ITS sequence between Uncaria sinensis rDNA and the existing Uncaria sinensisr DNA sequence in Genbank was 99.7% In the phylogenetic tree side by side clustered into one. Conclusion: Based on the sequence analysis of rDNA ITS region and the molecular biological method of phylogenetic tree construction, the accurate molecular identification of this genus Uncaria can be carried out, and the identification of species and the taxonomic status of the genus Uncaria Theoretical basis of molecular biology.