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应用ITS2条形码序列片段对我国市场药用植物栝楼与其混淆品共8份材料进行分子鉴定.对所有材料根部进行DNA提取纯化、PCR扩增并双向测序得到ITS2序列,将所得8条序列与Genbank中7条序列用Clustal X软件进行比对,BioEdit软件人工校正;利用MEGA5.0软件计算种内、种间遗传距离,并采用K2P距离法构建NJ和ML树,评价序列的鉴定效果.中药材植物各物种内遗传距离变异区间为0~0.077 9,平均为0.019,物种间遗传距离变异区间为0.004~0.594,平均0.436,种间距离明显大于种内距离;构建药用植物栝楼各物种NJ和ML系统进化树,二者结果一致,各物种均聚为一支,形成单系类群,支持率均在50%以上.ITS2条形码序列能够快速准确地从种的水平鉴定药用植物栝楼真伪.
ITS2 barcode sequence fragments were used to identify eight medicinal plants in China and eight samples of their adulterants.All DNAs were extracted and purified from all the roots of the material and PCR amplified and sequenced to obtain ITS2 sequences. Seven sequences were aligned by Clustal X software and BioEdit software was manually calibrated. Genetic distances between species and species were calculated using MEGA 5.0 software, and NJ and ML trees were constructed by K2P distance method to evaluate the sequence identification. The genetic variation range of each species was 0 ~ 0.077 9, with an average of 0.019. The variation range of genetic distance between species was 0.004 ~ 0.594 with an average of 0.436, and the interspecies distance was significantly greater than that within species. And ML phylogenetic tree, the two results are consistent, the species are clustered together to form a single line group, the support rate of more than 50% .ITS2 barcode sequence can quickly and accurately identify medicinal plants from the level of real Pseudo.