ISSR-PCR在石斛种间鉴别中的应用

Application of ISSR-PCR to identification of different Dendrobium Sw. species

目的 采用ISSR-PCR方法对石斛属9种植物进行鉴别,探讨不同种石斛在DNA分子水平上的差异。方法 选取10条由SSR组成的引物,对9种石斛进行PCR扩增及琼脂糖凝胶电泳分析。结果 10条引物中有7条扩增出多态性条带。每条引物可检测的多态性位点最少7个,最多14个,扩增片段大小为220-1260 bp。其中,引物UBC-807和UBC-864具有较高的多态性条带比率,均可以独立将所有被测种区分开来。结论ISSR-PCR作为一种简便、可靠的分子标记鉴定技术,可以作为石斛属种间鉴别的方法之一。

Objective To identify nine species of Dendrobium Sw. by inter-simple sequence repeats-polymerase chain reaction (ISSR-PCR) method and to investigate the difference among the species of Dendrobium Sw. at DNA molecular level. Methods Ten primers constituted by simple sequence repeats (SSR) were tested for PCR and sepharose electrophoresis of the nine species. Results Seven of the ten primers amplified polymorphic bands. The number of polymorphic band positions of each primer ranged from 7 to 14, and the size of fragments ranged from 220 to 1 260 bp. The amplification patterns of two primers, namely UBC-807 and UBC-864, were higher in terms of polymorphic and amplified band ratio. Each of them was able to distinguish all the examined species. Conclusion ISSR-PCR provides a quick, reliable molecular marker technique for identification of different species of Dendrobium Sw.