兼性无融合生殖龙须草SSR引物开发及杂交后代的检测

以来自中国5个省份12个居群的龙须草为材料,通过锚定PCR开发复合SSR引物、数据库检索和近缘物种SSR引物的引用等3种方法开发龙须草的SSR引物.结果显示:在锚定PCR开发复合SSR位点所设计的33对引物中筛选到有效扩增引物13对,表现多态性的引物有6对;利用基因特异序列法得到多态性引物3个;所用18条同源物种引物中有83%的SSR引物在供试的12个龙须草居群中都有清晰的SSR条带,其中56%的引物表现出多态性.3种方法在龙须草12个居群中共筛选得到了19对多态性引物,多态性比率约为73%.利用筛选得到的SSR多态性引物对来自2个居群的杂交F1代进行检测,鉴别出只有母本带型的无融合生殖后代和具有双亲带型的杂交后代,在子代中还出现偏父本带型、亲本缺失带型和变异带型等,证明所开发的SSR引物可以揭示龙须草生殖方式的复杂性,适用于龙须草遗传分析及亲缘关系鉴定.

Development of SSR Markers and Detection of Hybrid Progenies from Facultative Apomictic Eulaliopsis binata

Eulaliopsis binata is a facultative apomictic resource in Poaceae.Three methods,anchored PCR amplification of compound SSRs,datebase-searching and transferability of SSRs from related species,were used to isolate microsatellites in the 12 populations of Eulaliopsis binata from 5 provinces in China.33 primer pairs were designed by anchor-PCR amplification of compound SSRs,of which 13 primer pairs could get efficiency bands,and finally 6 primer pairs of these were polymorphic ones.3 polymorphic primer pairs were developed based on datebase-searching.In addition,18 primer pairs were selected from the relative species to analyze.It showed that of 83% primer pairs had product in 12 populations,and 56% primer pairs presented polymorphism.As results,19 polymorphic primer pairs were obtained by three different development methods.To prove the applicability of these SSR primers,F1 progeny plants from two populations were analyzed.Besides apomictic and sexual progenies,there were offspring plants whose bands toward the paternal's,deficiency to parents and variation with parents.