Development And Application Of SNP Markers Based On Walnut Black Spot Resistance Gene

Walnut black spot is a kind of bacterial infection disease of walnut,which mainly damages walnut leaves,twigs and fruits.Walnut black spot disease will lead to a serious reduction in walnut yield,restricting the development of walnut industry.It is of great strategic significance to breed and popularize walnut varieties resistant to black spot disease.Walnut transcriptome sequencing this test in different disease resistance,on the basis of using bioinformatics analysis screening disease-resistant gene and SNP loci associated with walnut black rot,build walnut black rot resistance are differences between the groups,using AS-PCR technique of SNP loci alleles genotype testing,development and walnut black rot resistance genes related SNP molecular markers,and use the marker detection of different walnut varieties on walnut black rot resistance,the main results were as follows:(1)a total of 306,879 SNP sites were detected by transcriptome sequencing of walnut with different resistances,including 120829 SNPS located in the coding region,64,797 synonymous SNPS in the coding region,and 56,032 nonsynonymous SNPS.According to the differential gene analysis and gene function analysis of trans criptome sequencing data,47 Unigene genes were selected that might be related to walnut resistance to black spot,and 36 non-synonymous SNP sites in coding region were found to be different between tiehetao and xiangling in 47 genes.(2)the allele specific primers were screened and verified by the allele specific population of susceptible materials,and 1 SNP marker was screened from 36 pairs of alleles,which was located on Unigene gene c84804 g3,pos-300,and the SNP type was C/A.The SNP genotype of the high-resistance material at this site was A/A homozygous,the SNP genotype of the high-sensitivity material at this site was C/C homozygous,and the genotype of the intermediate performance variety(MR/MS)at this site was C/A heterozygous.(3)the specific primers were used to identify 44 germplasm resources collected from the modern agriculture research and development base of sichuan agricultural university and the national forest germplasm resources of walnut and hickory in dechang county,and to rapidly detect the resistance of different varieties(clones)to walnut black spot disease.There are 16 materials with high resistance,12 materials with high sensitivity and 16 materials with intermediate properties.New ideas and methods were provided for the identification of walnut disease resistance,and the foundation was laid for the breeding of new varieties resistant to black spot.