Development and Utilization of KASP Marker for Brassica napus Clubroot Resistance Breeding

doi:10.7668/hbnxb.20192042

Abstract

Abstract: In order to accurately and efficiently screen clubroot-resistant materials and improve the breeding efficiency of clubroot resistance in Brassica napus.We sequenced the Crr1 gene of the disease-resistant parent of Brassica napus and the corresponding homologous gene LOC103834349 in the susceptible parent to search for SNP sites.Aiming at the non-synonymous mutation sites on 1486-1487, a set of competitive allele-specific PCR(Kompetitive Allele Specific PCR, KASP) markers was developed to accurately detect the genotypes of susceptible clubroot.Using this KASP marker, 771 individual plants from 42 F₂ populations were tested for typing, of which 315 individual plants contained homozygous susceptible genotype GG, 322 individual plants contained homozygous resistant genotype AA, and 134 individual plants contained homozygous susceptible genotype AA. The strain contains heterozygous allele GA. The KASP typing of 125 strains in the F₂ population numbered 2033 was χ² tested. Among them, 30 strains were homozygous for resistance genotype AA, 33 strains were homozygous for susceptible genotype, and 62 strains were heterozygous genotype GA.Disease-resistant materials and susceptible materials met the theoretical value of 3:1, and the ratio of disease-resistant homozygous genotypes, disease-resistant heterozygous genotypes and disease-susceptible genotypes met the theoretical value of 1:2:1, explained that the disease resistance site was a dominant single-gene disease resistance site.Combined with field phenotype detection and identification, AA typing and heterozygous GA typing were both disease-resistant phenotypes, and GG typing were susceptible phenotypes. The KASP typing results were consistent with the field identification results.The comparison results indicated that the KASP marker correctly typed clubroot resistant plants, indicating that the KASP marker developed based on the Crr1 gene and the SNP site of LOC103834349 can be accurately and efficiently applied to the molecular marker-assisted selection breeding of the clubroot resistant materials in Brassica napus.

Key words: Brassica napus, Clubroot, KASP marker, Resistance breeding, Marker-assisted selection

CLC Number:

Q78
S562.03

ZHONG Tingting, GUO Shifen, LU Wenbin, XIAO Gang. Development and Utilization of KASP Marker for Brassica napus Clubroot Resistance Breeding[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(4): 184-190. doi: 10.7668/hbnxb.20192042.

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