SSR Molecular Markers And Genetic Diversity Analysis Rubus In Cold Region

Rubus,belong to Rosaceae,is a kind of highly heterozygous small berry,and most of them are exist in the wild forms.Because of the diversity and instability of genes,the reaserches on Rubus is backward compared to other berries.In order to analyze the genetic diversity of this species,this study screened 15 pairs of high efficiency and good versatility primers were screened from 40 pair of primers by the SSR molecular markers.At the same time,it were compared in the research that three ways of electrophorests and two methods of dyeing.And the the main results are as follows:1.15 pairs of primers were selected from 40 pairs of primer by using 12 samples.They can be used for SSR-PCR amplification of red raspberry,blackberry and some wild species.2.Compared three PCR detection methods and two staining methods,the polyacrylamide gel electrophoresis' cost is lower,but it is not suitable for large-scale,multi-batch experiments,and the capillary electrophoresis is more accurate.The time required for silver staining was longer than that for EB,and the efficiency of EB staining was higher.3.Using the 15 pairs of primers,a total of 130 samples were amplified.And 219 bands were amplified.The number of bands amplified was between 3~12,with an average of 12.17,and the polymorphism rate was 98.2%.The average number of alleles obtained was 1.6749,the average Nei's index was 0.4390,and the average Shannon index was 0.5665.All the 130 samples were amplified using 15 pairs of primers.The number of bands amplified per sample was 12~25,which shows 15 pairs of primers could identify the 130 samples.4.All 130 samples were analyzed by cluster analysis.The similarity coefficient was between 0.79~0.97,and the similarity coefficient was 0.815.All the samples were divided into four categories: the first category was red raspberry cultivation sample,Jiaohe wild sample,yellow raspberries,three black raspberry samples and the wild Rubus,the wild Rubus.The second group is black raspberry,black pearl and red raspberry.The third kind is the spiny red from Shenyang.The fourth group is Madawaska and Willamette two red raspberry samples.Finally,Huang Shumei,raspberry and wild samples were isolated.