通过改良RAPD技术获得黑毛石斛、蜂腰石斛和石仙桃的3条SCAR新标记

3 new SCAR markers for Dendrobium williamsonii Day et Rchb.f.,Dendrobium findlayanum E.C.Parish et Rchb.f.and Pholidota chinensis Lindl.by improved RAPD amplification

目的寻找快速区分黑毛石斛、蜂腰石斛和石仙桃的序列特征扩增区域(sequence characterized amplified regions,SCAR)新标记。方法在改良的随机扩增多态性DNA(random amplified polymorphic DNA,RAPD)的基础上对10个物种进行基因组DNA随机扩增、分子克隆和测序,分别设计了可鉴别3个物种的SCAR标记。结果成功筛选出3个SCAR标记,其相应的特异性标记分别为N2-12、A2-17和A6-33;其中SCAR标记N2-12在黑毛种属中扩增出约750 bp的特异性条带,SCAR标记A2-17在蜂腰石斛中扩增出约750 bp的特异性条带,SCAR标记A6-33在石仙桃种属中扩增出约600 bp的特异性条带。结论N2-12、A2-17和A6-33这3对特异性SCAR新标记能快速、准确的区分出黑毛石斛、蜂腰石斛和石仙桃这3个物种,为今后进行该中药道地药材样品的鉴定提供了实验依据。

Objective To find the SCAR(sequence characterized amplified regions)markers that can quickly distinguish Den-drobium williamsonii Day et Rchb.f.,Den.findlayanum Par.et Rchb.f.and Pholidota chinensis Lindl..Methods In this paper,based on random amplified polymorphic DNA(RAPD),we randomly amplified the genomic DNA of 10 species,using the method of molecu-lar cloning and sequencing,and got the SCAR markers which can identify the three species.Results Three specific markers(N2-12,A2-17,A6-33)were screened.The primer N2-12 amplified a specific band of about 750 bp in the species of Dendrobium williamsonii Day et Rchb.f.,A2-17 amplified a specific band of about 750 bp in the species of Den.findlayanum Par.et Rchb.f.,and A6-33 ampli-fied a specific band of about 600 bp in the species of Pholidota chinensis Lindl..Conclusion These three pairs of specific SCAR mark-ers could quickly and accurately distinguish the above three species,providing an experimental basis for the identification of this au-thentic Chinese medicinal plant in the future.