鸡血藤及其混伪品matK基因分析和分子鉴定

Comparative Analysis and Molecular Identification of matKGene From Spatholobus suberectus and Its Adulterants

以鸡血藤及其4种常见混伪品为试材,采用植物基因组DNA提取试剂盒提取鸡血藤样品总DNA,并采用matK基因通用引物扩增和测序,从GenBank获取鸡血藤4种混伪品的matK基因序列,采用DNAMAN、MEGA等生物软件比对分析序列差异,计算遗传距离并构建聚类树,研究matK基因对鸡血藤及其混伪品的鉴别效果。结果表明:获得的鸡血藤及其混伪品matK序列长度为690bp,平均GC含量为31.1%,共发现236个差异位点,占总碱基数的34.2%。8个鸡血藤样品matK序列在41bp处存在A-G碱基差异。鸡血藤与混伪品之间的种间变异大于鸡血藤样品内的变异。基于matK基因序列建立的聚类树能够明显地鉴别鸡血藤及其混伪品。matK基因可作为鉴别鸡血藤及其混伪品的标准DNA序列。

In order to investigate the efficiency of matKgene on identifying Spatholobus suberectus and its adulterants,S.suberectus and four common adulterants were used as materials in this study.Genomic DNA was isolated from fresh leaves and dry materials of S.suberectus,and matKgene was amplified and sequenced using apair of universal primers.matKsequences of the adulterants were downloaded from GenBank database.The sequences were aligned and analyzed to find the difference and then genetic distance were calculated and the clustering tree was constructed using biological software.The results revealed that matKsequences acquired fromS.suberectus and its adulterants contained 690 base pairs with an average GC content of 31.1%.The maximal intraspecific distances among S.suberectus were 0.001,whereas the interspecific distance between S.suberectus and its adulterants ranged from 0.114 to 0.288.The clustering tree based on matKsequences can intuitively discriminate S.suberectus and its adulterants.matK gene can be used as a standard DNA sequence to distinguish Spatholobus suberectus and its adulterants.