甘蓝型油菜抗根肿病KASP标记开发和利用  

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

作　　者：钟婷婷 郭诗芬 卢文斌 肖钢[1,2] ZHONG Tingting;GUO Shifen;LU Wenbin;XIAO Gang(College of Agronomy,Hunan Agricultural University,Changsha 410128,China;Key Laboratory of Hunan Province for Rice and Rape Disease Resistance Breeding,Changsha 410128,China)

机构地区：[1]湖南农业大学农学院,湖南长沙410128 [2]水稻油菜抗病育种湖南省重点实验室,湖南长沙410128

出　　处：《华北农学报》2021年第4期184-190,共7页Acta Agriculturae Boreali-Sinica

基　　金：“十三五”国家重点研发项目(2016YFD0100500)。

摘　　要：为了准确高效的筛选抗根肿病材料,提高甘蓝型油菜根肿病抗性育种效率。通过对甘蓝型油菜抗病亲本的Crr1基因与感病亲本中相应的同源基因LOC103834349进行测序,寻找SNP位点,针对第1486-1487上的非同义突变位点,开发了一套精准检测抗感根肿病基因型的竞争性等位基因特异性PCR(Kompetitive Allele Specific PCR,KASP)标记。利用该KASP标记对来自42个F2群体的771个单株进行分型检测,其中315个单株含有纯合感病基因型GG,322个单株含有纯合抗病基因型AA,134个单株含有杂合等位基因型GA。其中对编号为2033的F2群体中125株材料的KASP分型情况进行χ^(2)检测,其中纯合抗病基因型AA 30株,纯合感病基因型GG 33株,杂合基因型GA 62株,经χ^(2)检测,抗病材料与感病材料符合3∶1理论值,抗病纯合基因型、抗病杂合基因型与感病纯合基因型的比值符合1∶2∶1理论值,说明该抗病位点为显性单基因抗病位点。结合田间表型检测鉴定,AA分型和杂合的GA分型单株田间检测均为抗病表型,GG分型均为感病表型,KASP分型结果与田间鉴定结果一致。比对结果说明该KASP标记对根肿病抗、感植株进行正确分型,表明基于Crr1基因和LOC103834349的SNP位点开发的KASP标记可以准确高效的应用于油菜抗根肿病材料的分子标记辅助选择育种。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 wasχ^(2) 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

关 键 词：甘蓝型油菜 根肿病 KASP标记 抗性育种 分子标记辅助选择 

分 类 号：Q78[生物学—分子生物学] S562.03[农业科学—作物学]