利用剩余杂合株系定位油菜根系发育主效QTL位点RT.C08  

Mapping of major QTL RT.C08 for root development in Brassica napus by using residual heterozygous lines

作　　者：黄林涛 沙爱华[1] 王新发[2] 顿小玲[2] 匡列琼 HUANG Lin-tao;SHA Ai-hua§;WANG Xin-fa;DUN Xiao-ling;KUANG Lie-qiong(Yangtze University,Jingzhou 434025,China;Oil Crops Research Institute of Chinese Academy of Agricultural Sciences,Key Laboratory of Biology and Genetic Improvement of Oil Crops,Ministry of Agriculture and Rural Affairs,Wuhan 430062,China)

机构地区：[1]长江大学,湖北荆州434025 [2]中国农业科学院油料作物研究所/农业农村部油料作物生物学与遗传育种重点实验室,湖北武汉430062

出　　处：《中国油料作物学报》2023年第2期260-270,共11页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金面上项目(31970526)。

摘　　要：油菜根系发育主效QTL的发掘与利用,对明确油菜根系发育机制,从根系形态建成的角度为油菜高产育种提供指导具有重要意义。本研究前期通过对一个重组自交系(RIL)群体(中双11×4D122)的QTL扫描,获得了1个控制根系发育的主效QTL位点RT.C08,最高可解释14.1%的根总体积表型变异。在此基础上,利用SNP芯片分型数据挑选出7个覆盖整个区间的剩余杂合单株,并以其构建RHL-F_(2)和RHL-F_(3)分离群体用作定位群体。基于SNP芯片数据和亲本序列差异,本研究在RT.C08的初定位区间开发了10个竞争性等位基因特异性(KASP)标记用于定位群体的基因分型,结合表型和基因型分析,发现剩余杂合系R53、R107和R260分离获得的B基因型(4D122基因型)株系在地上生物量、根鲜重、总根长、根表面积和根体积上优于A基因型(中双11号基因型)株系,这与RT.C08位点增效基因来自4D122一致,其它剩余杂合系未出现这种差异。进一步利用R53和R107的交换株系R53-F_(3)-5和R107-F_(3)-8,最终将RT.C08区间锁定在KASP标记NC8-6和C8-24663之间,且仍有三个KASP标记NC8-7、C8-1和C8-24013与其共分离,该区间在甘蓝型油菜Darmor参考基因组的物理距离约2.24 Mb。利用亲本重测序数据和基因表达谱分析,在此区间内共筛选出66个候选基因,其中BnaC08g21500D(CYCLIN D3)等12个基因已被报道调控根、叶片和/或下胚轴的生长。本研究为油菜优异根系分子标记辅助选择育种和重要基因的克隆奠定了良好的基础。The discovery and utilization of major QTLs for root development in rapeseed is of great significance to clarify the mechanism of root development and provide guidance for high-yield breeding in rapeseed from the per⁃spective of root morphogenesis.In the previous study,a major QTL locus RT.C08 controlling root development was obtained by QTL scanning in a recombinant inbred line(RIL)population(ZS11×4D122),which could explain up to 14.1%of the total root volume phenotypic variation.On this basis,7 residual heterozygous lines(RHLs)cover⁃ing the whole located interval were selected by SNP genotyping,and F_(2) and F_(3) segregation populations of these RHLs were constructed for further mapping.Ten kompetitive allele specific PCR(KASP)markers in the RT.C08 interval were developed according to the SNP and InDel differences between two parents.Combined with phenotyp⁃ic and genotypic analysis,it was found that in the separation lines of RHLs R53,R107 and R260,lines with geno⁃type B(4D122 genotype)were superior to lines with genotype A(Zhongshuang 11 genotype)in shoot fresh weight(SFW),root fresh weight(RFW),total root length(TRL),total root surface area(TSA)and total root volume(TRV).Consistent with the previous result,the synergistic allele of RT.C08 were derived from the parent 4D122.Other RHLs did not appear this difference.Using recombinant lines R53-F_(3)-5 and R107-F_(3)-8,the RT.C08 locus was further located between KASP markers NC8-6 and C8-24663,this locus was co-segregated with three mark⁃ers,namely NC8-7,C8-1 and C8-24013.According to the B.napus Darmor genome,the physical interval was about 2.24 Mb.Based on the resequencing data and gene expression profile analysis,66 candidate genes were screened in this interval,and 12 genes such as BnaC08g21500D(CYCLIN D3)have been reported to regulate root,leaf and/or hypocotyl growth.This study laid a foundation for molecular marker assisted selection breeding and cloning of important genes for excellent root system in rapeseed.

关 键 词：甘蓝型油菜 根系 剩余杂合系 QTL定位 KASP标记 

分 类 号：S565.4[农业科学—作物学]