基于EST-SSR标记鉴别园参、林下参和野山参的研究

Study on Authentication of Panax ginseng Cultivated in Gardens,Forest and Wild Panax ginseng by EST-SSR Markers

目的:分析EST数据库中园参、林下参和野山参的简单重复序列(SSR)位点多态性特征及规律,构建其SSR指纹图谱,探讨EST-SSR标记用于鉴别人参类药材的可行性。方法:应用生物信息学筛选出8对具有多态性的EST-SSR标记并设计引物,通过非变性聚丙烯酰胺凝胶电泳和毛细管电泳2种方法检测其扩增产物,比较其在园参、林下参和野山参中的分布特征。结果:发现SSR的多态性在园参、林下参和野山参中呈现逐步递减的趋势,这可能与遗传多样性、生长年限和生长环境相关。筛选出2对EST-SSR标记的扩增条带可以特异性的鉴别园参、林下参和野山参。所构建的系统发育关系表明,与园参相比,林下参和野山参的亲缘关系较近。结论:SSR分子标记可以用于园参、林下参和野山参的鉴定,且结果准确可靠,具有重要的应用价值。

Objective:To analyze the polymorphism characteristics and rules of simple repeat sequence(SSR)loci of Panax ginseng cultivated in gardens,forest and wild Panax ginseng in EST database,to construct SSR fingerprint,and to explore the feasibility of EST-SSR markers for identification of Panax ginseng.Methods:Eight EST-SSR markers with polymorphism were screened by bioinformatics and primers were designed.The amplified products were detected by non-denaturement polyacrylamide gel electrophoresis and capillary electrophoresis,the distribution characteristics of the amplified products in Panax ginseng cultivated in gardens,forest and wild Panax ginseng were compared.Results:SSR polymorphism showed a decreasing trend in Panax ginseng cultivated in gardens,forest and wild Panax ginseng,which might be related to genetic diversity,growth years and growth environment.Two amplification bands of EST-SSR markers were screened out to identify Panax ginseng cultivated in gardens,forest and wild Panax ginseng.The phylogenetic relationship established showed that the Panax ginseng cultivated in forest and wild Panax ginseng were more closely related than the Panax ginseng cultivated in gardens.Conclusion:SSR markers can be used to identify Panax ginseng cultivated in gardens,forest and wild Panax ginseng,and the results are accurate and reliable,which have important application value.