[关键词]
[摘要]
目的以核糖体转录间隔区(rDNA ITS)序列为分子标记,对钩藤属的2种常用中药材毛钩藤Uncariahirsute和无柄果钩藤U.sessilifructus进行遗传分析,阐明钩藤属内不同种间的分子系统发育关系。方法通过改良CTAB法对毛钩藤和无柄果钩藤进行总DNA提取,利用通用引物对rDNA ITS序列进行PCR扩增,RFLP分析和序列测定,用PAUP软件进行系统发育分析,构建最大简约树。结果获得毛钩藤和无柄果钩藤的限制性内切酶(MspⅠ&HaeⅢ)酶切图谱以及rDNA ITS区的完整序列,其序列长度均为718 bp。结论通过基于rDNAITS序列进行RFLP分析、序列测定和系统发育树构建的分子生物学方法,能够对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依据。
[Key word]
[Abstract]
Objective In order to clarify the phylogenetic relationships between the different species of Uncaria Schred,genetic analysis was carried out for Uncaria hirsute and U.sessilifructus by using the rDNA ITS sequence as molecular marker.Methods Total DNA was extracted with modified CTAB method and thereby rDNA ITS regions were amplified with common primers.The rDNA ITS amplicon was characterized by RFLP,sequencing and phylogenetic analyses using PAUP by maximum parsimony.ResultsThe Msp Ⅰ-and Hae Ⅲ-based RFLP patterns of rDNA ITS from U.hirsute and U.sessilifrctus were obtained,where the resultant entire sequences of rDNA ITS region were both 718 bp.Conclusion Based on molecular biology methods of rDNA ITS sequence analysis,accurate molecular identification could be performed on traditional Chinese medicinal material plants in Uncaria Schred and provide molecular evidence for taxonomy and identification of different species in Uncaria Schred.
[中图分类号]
[基金项目]
国家自然科学基金资助项目(30800227)