水稻矮秆多分蘖突变体的表型分析及基因克隆

Phenotypic Analysis and Gene Cloning of A Dwarf and More Tillers Mutant in Rice

  • 摘要: 以粳稻品种空育131为材料,经EMS诱变获得一个株型突变体,其表型为植株矮化、分蘖增加及粒型较小等,命名为dmt(dwarf and more tillers)。遗传分析表明,突变体表型受一对隐性核基因所控制。利用独脚金内酯的类似物GR24处理dmt突变体能够恢复野生型表型,说明dmt突变可能与独脚金内酯合成途径相关。利用dmt与空育131构建回交群体,基于BSA和MutMap相结合的方法,将突变位点定位于第4染色体的LOC_Os04g46470位点,测序表明,该基因在第5个外显子最后一个碱基发生了G到A的突变,使该基因错误剪切,104 bp的第5内含子被转录,导致该基因移码突变。LOC_Os04g46470编码胡萝卜素裂解双加氧酶7(carotenoid cleavage dioxygenase 7,CCD7),是独脚金内酯合成途径的基因。此外,突变体中CCD7的表达量高于对照,也证明dmt中独脚金内酯合成途径突变,造成负反馈抑制减弱。

     

    Abstract: A dwarf and more-tiller mutant (dmt) was obtained from the ethyl methanesulfonate (EMS) mutant library of Kongyu131 (Oryza stativa ssp Japonica cultivar). The dmt mutant exhibits decreased plant height, more tillers and smaller grains. Genetic analysis indicated that the dmt phenotype is caused by a single recessive mutation. The Strigolactones (SLs) analogue GR24 could restore the dmt phenotype, indicating that SLs biosynthesis pathway of dmt was disrupted. Combining bulked segregant analysis (BSA) and MutMap analysis, LOC_Os04g46470 encoding a carotenoid cleavage dioxygenase 7 (CCD7) was identified as a candidate gene for the dmt. Sequence analysis revealed a G-to-A transition point mutation at the last base of the fifth exon of LOC_Os04g46470 in the dmt, resulting in a frame shift with the fifth intron transcribed. Moreover, the CCD7 transcript level in the dmt mutant was significantly higher than that in the WT, supporting the negative feedback role of CCD7 in SLs biosynthesis regulation.

     

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