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通过EMS诱变获得遗传稳定的水稻褐色颖壳突变体(brown hull 6,简称bh6)。与野生型IR64相比,在自然条件下突变体的颖壳颜色自抽穗期开始逐渐加深直至蜡熟期颖壳呈深褐色,除千粒重和穗长显著降低外,其他农艺性状(株高、每株有效穗数、每穗实粒数、结实率)未发生显著改变。遗传分析表明,该褐色颖壳性状受一对隐性核基因控制。构建突变体bh6与正常色颖壳水稻CPSLO17的F2分离群体作为基因定位群体,将该基因精细定位在第9染色体长臂上106kb的区段内。序列分析发现,突变体中F-box蛋白基因OsFBX310(LOC_Os09g12150)编码区第1013位碱基由G突变为A,致使其所编码的蛋白序列第338位的甘氨酸(Gly)突变成天冬氨酸(Asp)。通过构建OsFBX310功能互补载体进行转化试验,结果表明,突变体转基因植株的谷壳颜色恢复为正常颜色。该基因是已报道的水稻OsFBX310基因的新等位基因,命名为OsFBX310bh6。OsFBX310bh6及其野生型等位基因的克隆对深入研究338位甘氨酸/天冬氨酸在类黄酮代谢过程中的作用,以及丰富水稻特异种质资源具有积极意义。
A genetically stable brown hull mutant (brown hull 6, bh6) was obtained by EMS mutagenesis. Compared with wild-type IR64, the glume color of the mutant under natural conditions gradually deepened from the heading stage to the dark brown color at the ripening stage. Except for the 1000-grain weight and ear length decreased significantly, the agronomic traits (plant height, Effective panicles per panicle, seed number per panicle, seed setting rate) did not change significantly. Genetic analysis showed that the brown glume trait was controlled by a pair of recessive nuclear genes. The F2 segregating population of the mutant bh6 and the normal chromophorous rice CPSLO17 was constructed as a genetic locus, and the gene was finely mapped on the 106 kb segment of the long arm of chromosome 9. Sequence analysis revealed that the base of the 1013 base of the F-box protein gene OsFBX310 (LOC_Os09g12150) in the mutant was mutated from G to A, resulting in the mutation of glycine (Gly) at position 338 of the F-box protein gene in the mutant into aspartate Asp). By constructing OsFBX310 functional complementation vector for transformation test, the results showed that the chaff color of the mutant transgenic plants returned to normal color. This gene is a reported new allele of rice OsFBX310 gene, named OsFBX310bh6. Cloning of OsFBX310bh6 and Its Wild-type Alleles It is of great significance to further study the role of 338 glycine / aspartate in flavonoid metabolism and to enrich rice-specific germplasm resources.