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目的:建立滇皖产区铁皮石斛SSR指纹图谱,并构建不同居群的SSR分子身份证。方法:使用生物信息学方法分析铁皮石斛(Dendrobium officinale)全基因组SSR(Simple sequence repeat)序列,获得SSR位点,使用Primer Premier 5.0软件设计引物并进行PCR扩增,选择高多态性引物进行遗传多样性分析并建立SSR指纹图谱代码,使用二维码生成器表述铁皮石斛居群的分子身份证。结果:共获得91 653个SSR位点,平均分布距离为15.44 kb。据此开发了75对SSR引物,筛选出32对多态性丰富、带型清晰的引物,对11个滇、皖产区的铁皮石斛居群进行遗传多样性分析,共得到117个多态信息位点,平均每个SSR引物多态信息含量(PIC)为0.62,变化范围为0.22~0.80;铁皮石斛不同居群进行亲缘关系聚类表明11个铁皮石斛居群可分为三支,与种质来源分布相一致。筛选出4对多态性高、带型清晰易于统计的引物组合,建立不同居群铁皮石斛的SSR指纹图谱库和不同居群的分子身份证。结论:SSR分子身份证可为今后铁皮石斛的品种鉴别提供准确、可靠的方法和溯源信息平台。
Objective: To establish SSR fingerprinting of Dendrobium officinale from Yunnan, Anhui and to construct SSR molecular ID cards of different populations. Methods: The whole genome sequence of Dendrobium officinale was analyzed by bioinformatics method to obtain SSR loci. Primer Premier 5.0 software was used to design primers and PCR amplification, and high polymorphism primers were selected for inheritance Diversity Analysis and the establishment of SSR fingerprinting code, the use of two-dimensional code generator expression Dendrobium populations molecular ID. Results: A total of 91 653 SSR loci were obtained, with an average distribution distance of 15.44 kb. Based on this, 75 pairs of SSR primers were developed and 32 pairs of polymorphic primers with clear bands were screened. Genetic diversity among populations of Dendrobium officinale from 11 areas in Yunnan and Anhui Provinces was analyzed. A total of 117 polymorphic loci The polymorphism information content (PIC) of each SSR primer was 0.62 and the variation range was 0.22 ~ 0.80. Cluster analysis showed that 11 populations of D. officinale were divided into three groups, The source of the distribution of the same. Four pairs of primer combinations with high polymorphism and clear bands were screened, SSR fingerprinting library and molecular ID cards of different populations were established. Conclusion: The SSR molecular ID card can provide an accurate and reliable method and traceability information platform for the identification of Dendrobium candidum varieties in the future.