珍稀铁皮石斛SSR标记的开发及种质纯度鉴定  

Development of microsatellites of Dendrobium officinale and its application in purity identification of germplasm

作　　者：谢明璐[1] 侯北伟[1] 韩丽[1] 马艳红[1] 丁小余[1] 

机构地区：[1]南京师范大学生命科学学院,江苏南京210046

出　　处：《药学学报》2010年第5期667-672,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(30870234);国家基础人才培养基金资助项目(J0730650);江苏省自然科学基金资助项目(BK2008431)

摘　　要：本研究利用磁珠富集法开发了60对铁皮石斛的SSR引物,从中筛选出15对多态性位点丰富、带型清晰、重复性好的引物。利用筛选出的SSR引物对铁皮石斛野生居群材料进行了遗传分析,共鉴定出等位变异92个,平均每个SSR位点等位变异数为6.1个;表观杂合度(HO)范围是0.60～0.85,平均值为0.72;期望杂合度(HE)范围是0.49～0.85,平均值为0.74。平均每个SSR位点多态信息含量(PIC)为0.702,变化范围为0.437～0.829。利用15对SSR引物对20种石斛试验材料进行跨种扩增,检测出有13对SSR引物具有种间通用性。此外,还利用所筛选的4对SSR引物检测了集约化种植过程中的铁皮石斛组培苗的种质纯度,结果显示:所开发的SSR标记可用于铁皮石斛组培苗的品种纯度鉴别。Simple sequence repeat （SSR） was used to investigate the genetic diversity and structure of Dendrobium officinale.A total of 15 primer pairs with stable and repeatable polymorphism were screened out from 60 SSR primer pairs developed by the method of microsatellite enrichment by magnetic beads.Forty-eight samples of Dendrobium officinale were analyzed in genetic polymorphism.These loci were polymorphic and displayed 3 to 9 alleles per locus with a mean number of 6.1.The observed and expected heterozygosities ranged from 0.60 to 0.85 and from 0.49 to 0.85 respectively.The polymorphic information content （PIC） of each SSR locus varied from 0.437 to 0.829 with an average of 0.702.Fifteen primer pairs were used in Dendrobium cross-species amplification and totally 13 primer pairs were proved to have the transferability in D.officinale related species.In addition,500 tissue culture plantlets of D.officinale were tested for purity identification by means of PCR amplification with four SSR primer pairs.The results showed that SSR technique is a feasible,simple and inexpensive method for determining adulterants in germplasm identification.

关 键 词：铁皮石斛 微卫星标记开发 通用性检测 纯度鉴定 

分 类 号：R931.5[医药卫生—生药学]