Construction Of DNA Fingerprinting And Analysis Of Genetic Diversity For Common Bean

Phaseolus vulgaris also known as green beans,Meidou,leguminous,belong to the common species of genus bean,which has the largest planting area of edible beans in the world.Phaseolus vulgaris resources are rich in our country,the effective utilization of germplasm resources still need further research,So the understanding of the genetic basis of Phaseolus vulgaris varieties,master the genetic relationship among populations of Phaseolus vulgaris,Determine protection ways and means,its are important for Phaseolus vulgaris varieties identification and traceability information summary.This study based on SSR molecular marker technology,screened 11 pairs core primers,On the analysis the genetic diversity of 99 kidney bean,clear the genetic relationship between the material,from the DNA level to identify Phaseolus vulgaris varieties.The purpose of this experiment as the Phaseolus vulgaris standard fingerprints for libraries to provide theoretical basis.The main research results are as follows:1.With 2 primers screening,selected 11 core primers of relatively high polymorphism,good repeatability,Each pair of primers average PIC value is 0.649,and average shannon diversity index is 1.3029,indicate that the screened primer is efficient primers.Through clustering analysis can see 11 core primers can completely discriminat 36 kidney bean varieties.2.Based on SSR fluorescence labeling analysis of genetic diversity by using 11 pairs of core primers for 9 different geographical origin of the 99 varieties,127 alleles were detected,7?19 alleles were detected by each pair of primers.The average number is 11.54.The value of polymorphism information of core primers were 0.5302?0.8686,with an average of0.7035.The result showed that the primers had a high polymorphism and the ability to distinguish.Cluster analysis results showed that the 99 types of kidney bean genetic similarity coefficient were 0.23?0.98,the diversity of population genetic diversity among different provinces was better,and the genetic diversity of Heilongjiang population was more abundant.3.99 copies of kidney bean fingerprint was constructed by 11 pairs of core primers,the fingerprint of each material had been unique and built a set of fingerprint code using the fingerprint encoding form,it can intuitively judge varieties of homozygous and heterozygous type.Of which 8 pairs of primers BM139,BM140,BM152,BM156,BM160,BM164,GATS91,BM157 had unique characteristics.SSR markers were suitable for the construction of common bean varieties of the DNA fingerprint database,it can provide a technical support for the future identification of common bean varieties.4.The method of SSR-PCR product was compared with the conventional silver staining and fluorescence detection,8 pairs of primers with good polymorphism and specificity were selected,genetic diversity analysis of 20 varieties in Heilongjiang province using silver staining and fluorescence detection.Both methods can be used to identify the tested varieties.Fluorescence detection method can detect the number of alleles with silver staining method can be more accurated,with similar genetic background to distinguish.As a result,the genetic diversity among the cultivars reflected by the fluorescence detection method were slightly higher than that of the silver staining method.