• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

利用BSA-Seq方法鉴定谷丰B抗稻瘟病基因

BSA-Seq Identification of Blast-resistance Genes in Gufeng B Rice

  • 摘要:
      目的  挖掘和鉴定谷丰B稻瘟病抗性基因,了解谷丰B稻瘟病抗性遗传模式。
      方法  以谷丰B和日本晴杂交获得F1和F2代遗传群体,接种稻瘟菌不同生理小种并分析抗病遗传模式;在F2群体中挑选极端抗/感单株构建DNA混合池,利用群体分离分析法(BSA)定位关联区域。
      结果  谷丰B对KJ201、RB22、CHNOS、RB6、2Y838-1、501-3和IR16-1等菌株均表现高抗性,表明谷丰B基因组可能携带了广谱高抗稻瘟病基因。谷丰B和日本晴杂交,F1群体表现抗501-3和IR16-1,F2群体的抗病/感病分离比不符合3 1,推测谷丰B基因组存在多个位点影响稻瘟病抗性。对F2群体的极端抗病、感病混合池及亲本DNA进行全基因组测序,鉴定了1 756 964个单核苷酸多态性(SNPs)标记。分析子代△SNP-index,定位到2个与抗病性显著关联区间,分别为Chr.6: 10 082-11 397 kb和Chr.11: 120-266 kb。其中,6号染色体的关联区间与Pi2/9抗病位点等位,区间内含有4006个SNPs和623个插入缺失(InDels)标记;11号染色体的关联区间含有752个SNPs和195个InDels标记。
      结论  谷丰B对强致病力501-3菌株抗性可能是由第6号和11号染色体上的基因共同控制。研究结果为谷丰B抗性基因的精细定位及基因克隆奠定了基础,并为水稻抗稻瘟病分子标记辅助选择提供标记资源。

     

    Abstract:
      Objective  The rice cultivar Gufeng B confers strong, broad-spectrum, durable resistance against various rice blast isolates. The present study was aim to identify and map the blast resistance gene(s) in Gufeng B.
      Method   The F1 and F2 population were obtained by crossing Gufeng B and Nipponbare, and the genetic model of blast resistance was analyzed after inoculating with 7 strains of Magnaporthe grisea. Subsequently, F2 population was used to construct a resistant pool and a sensitive pool respectively, and to map the associated loci via the method of bulked segregation analysis.
      Result   Gufeng B exhibited high resistance to all of the tested strains, such as KJ201, RB22, CHNOS, RB6, 2Y838-1, 501-3 and IR16-1, suggesting that Gufeng B may carry the broad-spectrum and high resistance genes. The F1 progenies from the cross between Gufeng B and Nipponbare conferred resistance against the strains 501-3 and IR16-1, and the segregation ratio of resistance and susceptibility among F2 progenies does not fit 3:1, assuming that the resistance against the strains 501-3 and IR16-1 were controlled by multiple locus in Gufeng B. Whole genome re-sequencing of the two parental lines Gufeng B and Nipponbare identified 1,756,964 SNPs. Calculation results of △SNP-index showed that there were two candidate loci conferring resistance to rice blast disease, which were located at Chr.6: 10,082-11,397Kb, corresponding to the Pi2/9 locus, and Chr.11: 120-266Kb. 4006 SNPs and 623 InDels markers were searched within the interval of Chromosome 6, 752 SNPs and 195 InDels within the corresponding region of Chromosome 11, respectively.
      Conclusion  The resistance of Gufeng B to 501-3 strain may be controlled by two resistance genes on chromosomes 6 and 11. Our results laid the foundation for finely mapping and cloning the resistance genes in Gufeng B, and provided marker resources for molecular marker-assisted selection.

     

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