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目的建立一种准确、快速的凉茶药材凉粉草与其混伪品的DNA条形码鉴定方法。方法本实验收集凉粉草及其混伪品共5个物种和4个变种的55份样本,提取基因组总DNA,通过PCR扩增和Codon Code Aligner软件拼接注释获得完整的ITS2序列,然后应用种内种间K2P遗传距离和系统进化NJ树分析进行物种鉴定。结果凉粉草ITS2序列长度为232 bp,种内遗传距离为0,远小于其与混伪物种间的最小遗传距离0.210 2~0.310 9;NJ树结果表明,凉粉草独成一支,与其混伪品均可准确区分。结论基于ITS2序列的DNA条形码技术能够准确有效地实现凉粉草与其混伪品的快速鉴别,保障凉茶药材凉粉草的使用安全。
Objective To establish an accurate and rapid DNA barcode identification method for Herbal Medicine jelly grass and its adulterants. Methods In this study, we collected 55 samples of 5 species of Grifola frondosa and its adulterants and 4 variants. The total genomic DNA was extracted. The complete ITS2 sequence was obtained by PCR amplification and splicing with Codon Code Aligner software. Interspecific K2P genetic distance and phylogenetic tree NJ analysis for species identification. Results The sequence length of ITS2 was 232 bp and the intraspecific genetic distance was 0, which was much smaller than the minimum genetic distance between the two species and the pseudo-pseudo-species ranged from 0.210 2 to 0.310 9. The results of NJ tree showed that the genus Ginkgo grew independently, Can be accurately distinguished. Conclusion DNA barcoding based on ITS2 sequence can accurately and effectively identify jelly grass and its adulterants and ensure the safe use of herbal medicine.