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目的:采用ISSR分子标记技术对中国药典所列秦艽的4种基原植物秦艽、麻花秦艽、粗茎秦艽和小秦艽及其近缘种黄管秦艽、管花秦艽进行分子鉴定和亲缘关系分析,以期在DNA分子水平上为黄管秦艽和管花秦艽作为秦艽替代品使用的科学性和合理性提供一定的参考。方法:筛选出8条重复性好、扩增条带清晰的引物用于所有样品的PCR扩增和琼脂糖凝胶电泳分析,使用NTSYSpc-2.10E软件计算相似系数并绘制聚类图。结果:ISSR指纹图谱共检测到9条种间特异性条带,可用来鉴定除小秦艽以外的5个种的秦艽;秦艽替代品黄管秦艽和小秦艽最先聚在一起,管花秦艽与小秦艽、麻花秦艽的亲缘关系更近,粗茎秦艽与其他5个种的秦艽存在较远的亲缘关系。结论:基于ISSR分子标记的聚类分析结果支持黄管秦艽和管花秦艽作为秦艽的替代品使用。
OBJECTIVE: To identify and analyze the genetic relationships of four species of Gentiana macrophylla, Gentiana macrophylla, Gentiana macrophylla, Gentiana macrophylla, Gentiana macrophylla, Gentiana macrophylla, Gentiana macrophylla, Gentiana macrophylla, Gentiana macrophylla, Gentiana macrophylla, Gentiana macrophylla, In order to provide some reference for the scientific and rationality of the use of Gentiana macrophylla and Gentiana macrophylla as substitutes of Gentiana macrophylla at DNA molecular level. Methods: Eight primers with good repeatability and clear bands were selected for PCR amplification and agarose gel electrophoresis analysis of all the samples. NTSYSpc-2.10E software was used to calculate the similarity coefficient and draw the cluster diagram. Results: Nine species-specific bands were detected by ISSR fingerprinting, which could be used to identify five species of Gentiana macrophylla in addition to Gentiana macrophylla. Gentiana macrophylla Qin Qin and Gentiana macrophylla were the first to be clustered together, The genetic relationship between Gentiana macrophylla and Gentiana macrophylla is closer, and Gentiana macrophylla has more distant kinship with other genus Gentiana genus. Conclusion: The results of cluster analysis based on ISSR molecular markers support the use of Gentiana macrophylla and Gentiana macrophylla as a substitute for Gentiana macrophylla.