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目的:对土茯苓基原植物及其近缘种进行psbA-trnH条形码鉴定。方法:提取样品DNA,利用PCR技术对样品进行叶绿体基因psbA-trnH片段扩增并双向测序。所得序列经CodonCode Aligner拼接后,利用MEGA 6.0软件进行数据分析;计算种内种间K2P遗传距离并采用邻接法构建NJ系统树。结果:土茯苓基原植物种内最大K2P遗传距离远小于种间最小K2P遗传距离;由构建的系统聚类树图可以看出,不同来源的土茯苓基原植物聚成一支,表现为单性系,并与其近缘种明显分开。结论:psbA-trnH序列作为DNA条形码可以有效地鉴别土茯苓基原植物及其近缘种,为土茯苓药材基原鉴定及临床安全用药提供了分子依据。
OBJECTIVE: To identify psbA-trnH barbodes from Tuckahoe plants and their related species. Methods: DNA samples were extracted and the chloroplast psbA-trnH fragment was amplified by PCR and sequenced bidirectionally. The obtained sequence was spliced by CodonCode Aligner, and the data were analyzed by using MEGA 6.0 software. The genetic distance of intraspecific K2P was calculated and the NJ phylogenetic tree was constructed by using adjacency method. Results: The maximum K2P genetic distance of Fusarium-based primordial species was far less than the minimum K2P genetic distance between species. According to the phylogenetic tree diagram, the Fusarium-based plants of different origins were clustered into one, Department, and its close relatives clearly separated. Conclusion: The psbA-trnH sequence can be used as a DNA barcode to identify Gentiana chinensis and its related species effectively. It provides a molecular basis for the identification of Gentiana macrophylla and clinical safe medication.