基于近缘物种SSR引物和EST-SSR序列的梅花SSR引物开发

Development of mei flower's SSR primers from closely related species' SSR and EST-SSR

为获得更多的梅花SSR引物,本研究筛选了84对梅花近缘物种的引物,并从梅花EST数据库的序列中设计了23对EST-SSR引物,用12个梅花品种对这些引物进行了多态性检测。结果表明,近缘物种的引物有69对能扩增出条带,有效扩增率达82.1%,从中筛选的14对引物共得到85个等位基因,平均值为6个。有效等位基因数在2.072～5.539之间,平均值为4。PIC多态信息指数在0.483～0.786之间,平均值为0.695。Shannon指数的平均值为1.528。12个品种遗传相似系数在0.13～0.60之间,平均值为0.26。‘玉蝶龙游’梅和‘小玉蝶’相似系数最大,为0.60;‘美人’梅和‘黄金’梅相似系数最小,为0.13。设计的EST-SSR引物有22对能扩增出条带,有效扩增率达95.6%,其中有11对引物表现出多态性,占全部EST-SSR的47.8%。

In this study,we analyzed 84 SSR primer pairs from closely related species and designed 23 EST-SSR primer pairs from NCBI for developing more mei flower（Prunus mume Sieb.et Zucc.）new SSR makers.Twelve mei flower cultivars were used to do polymorphic test on these primers.The results show that 69 SSR primer pairs can amplify bands with the efficient amplification ratio of 82.1%.A total of 85 alleles were detected by 14 primer pairs with an average of six effective alleles per primer.The effective number of alleles was between 2.072 and 5.539,with the average of 4.The polymorphic information content（PIC） value was between 0.483 and 0.786,with the average of 0.695.The average Shannon Index was 1.528,with the genetic similarity coefficient among 12 materials ranging from 0.13 to 0.60 and the average of 0.26.Based on the UPGMA analysis,P.mume ＇Yudie Longyou＇ and P.mume ＇Xiao Yudie＇ contained similar inheriting basement,about 0.60,and the similarity value between P.mume ＇Meiren＇ and P.mume ＇Huangjin＇ was smallest,about 0.13.The designed 22 EST-SSR primer pairs can amplify bands,with the effective amplificaition rate of 95.6%.Eleven primer pairs demonstrated the polymorphic characteristics,accounting for 47.8% of all the EST-SSR.