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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。
Taking Millettia and its four common adulterants as test materials, the genomic DNA extraction kit was used to extract the total DNA of Millettia samples and amplified and sequenced with matK gene universal primers. Four species of Millettia were obtained from GenBank The matK gene sequence of the adulterated product was analyzed by DNAMAN, MEGA and other biological softwares. The genetic distance was calculated and the clustering tree was constructed. The discriminating effect of matK gene on Millettia and its adulterants was studied. The results showed that the length of matK sequence was 690 bp and the average GC content was 31.1%. A total of 236 differential sites were found, accounting for 34.2% of the total base number. There were A-G base differences at 41bp for matK sequences of 8 samples of Millettia. The interspecific variation between Millettia and Dumbo was greater than that within Millettia samples. The clustering tree based on the matK gene sequence can clearly identify Millettia and its adulterants. matK gene can be used as a standard DNA sequence to identify Millettia and its adulterants.