山西省吉县刺槐无性系种质遗传多样性的EST-SSR分析

EST-SSR analysis of genetic diversity of Robinia pseudoacacia clones in Jixian County, Shanxi Province of northern China

【目的】了解各对EST-SSR引物检出的等位简单序列重复片段数量,解析吉县良种基地刺槐无性系种质的多态性状况与种质间亲缘关系,为引物利用和刺槐种质遗传管理提供参考。【方法】利用课题组开发的45对EST-SSR引物进行PCR扩增,使用毛细管电泳仪对产物进行检测,用Popgene 1.32版软件对群体遗传参数进行评估,利用NTsys 2.10e进行聚类分析。【结果】45对EST-SSR引物中,除了引物RP-24均表现出良好的多态性。44对引物共检测到等位重复序列298个,每对引物检测到2~15个,平均6.77个。群体观测杂合度(Ho)的变化范围为0.022 7~0.842 1,平均为0.412 6。期望杂合度(He)的变化范围为0.088 7~0.822 3,平均为0.484 0。引物多态性信息量(PIC)变化范围为0.085 2~0.796 6,平均为0.444 4。【结论】该基地的刺槐无性系种质具有较高的遗传多样性。由聚类结果可知,在遗传距离为0.81时,可将96份刺槐无性系种质聚成5类。聚类图在分子水平上显示了刺槐无性系种质间的亲缘关系,研究结果对于利用这些引物用于刺槐的种质评价、资源管理和辅助育种有积极的参考价值。

[Objective]This paper aims to understand the number of alleles of simple sequence repeats detected by EST-SSR primers,and to analyze the polymorphisms of germplasm of Robinia pseudoacacia clones in Ji xian County of Shanxi Province,northern China and the genetic relationship between germplasms,and provide reference for primer utilization and genetic management of Robinia pseudoacacia germplasm.[Method]45 pairs of EST-SSR primers developed by our laboratory were used for PCR amplification,and the products were detected by capillary electrophoresis,the genetic parameters of population were evaluated by Popgene 1.32 software,and cluster analysis was performed by NTsys 2.10 e.[Result]Except for primer RP-24,45 pairs of EST-SSR primers showed good polymorphism.A total of298 allelic repeat sequences were detected in 44 pairs of primers,and 2 to 15 were detected in each pair,with an average of 6.77.The observed heterozygosity(Ho)of the population ranged from 0.022 7 to 0.842 1,with an average of 0.412 6.The expected heterozygosity(He)varies from 0.088 7 to 0.822 3,averaging0.484 0.Primer polymorphism information(PIC)ranged from 0.085 2 to 0.796 6,with an average of 0.444 4.[Conclusion]Robinia pseudoacacia clonal germplasm in this base has high genetic diversity.It can be seen from the clustering results that 96 genetic clones of the hedgehog clones can be clustered into five classes when the genetic distance is 0.81.Cluster maps showed the genetic relationship among the clones of Robinia pseudoacacia at the molecular level,and the results of the study have positive reference value for the use of these primers for germplasm evaluation,resource management and assisted breeding of Robinia pseudoacacia.