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粒形是影响水稻产量的重要因子,对粒形相关基因的发掘和利用一直倍受重视。在已克隆的粒形基因中,GW2是一个控制粒宽的重要基因。通过同源克隆的方法,获得极端大粒水稻TD70和极端小粒水稻Kasalath的GW2基因cDNA片段,分析序列差异,设计功能标记,检测240个来源于TD70/Kasalath的重组自交系(RILs)和部分籼稻骨干恢复系、江苏省主栽粳稻品种的基因类型及其效应。结果表明,TD70的GW2与大粒粳稻WY3完全一致,与已知序列(GenBank:EF447275)相似性达99.77%,在碱基的第114位,T突变为C,但并不改变氨基酸性质;在第316位缺失碱基A,造成316位以后的所有碱基前移一位,氨基酸均发生变化,在碱基的第346、第347、第348位形成TAA的终止密码子,提前结束了TD70的GW2蛋白的表达。而Kasalath的GW2序列仅在碱基的第114位,由T突变为C,但并不改变氨基酸性质;GW2Kasalath的316位没有突变,表达没有提前终止;设计的dCAPs标记,产物经内切酶ApoI酶切,TD70为21bp和30bp两个片段,而Kasalath条带为51bp;240个RILs中含GW2TD70基因的株系有82个,含GW2Kasalath基因的株系有158个,两者在粒宽和粒重表型上存在显著差异;8个籼稻骨干恢复系和10个江苏省主栽粳稻品种中都不存在GW2TD70基因。GW2TD70基因对籽粒宽度有一定的调控作用,水稻育种中可以利用GW2TD70基因来提高籽粒粒重。
Grain shape is an important factor affecting the yield of rice. It has been paid much attention to the exploration and utilization of grain-related genes. Among the cloned grain genes, GW2 is an important gene controlling grain width. Through homologous cloning method, cDNA fragments of GW2 gene of extreme grain rice TD70 and extreme grain rice Kasalath were obtained, and sequence differences were analyzed. Functional markers were designed and 240 recombinant inbred lines (RILs) derived from TD70 / Kasalath and some indica rice Key Restorer Line, Genotypes and Effects of Main Japonica Rice Varieties in Jiangsu Province. The results showed that GW2 of TD70 was completely identical with WY3 of large grain japonica rice, and its similarity with known sequence (GenBank: EF447275) was 99.77%. At position 114 of the base, T was mutated to C, but did not change the nature of amino acids. The deletion of base A at position 316 resulted in a forward shift of all bases after position 316 and a change of amino acids. A stop codon of TAA was formed at positions 346, 347 and 348 of the base, leading to the termination of TD70 GW2 protein expression. However, the GW2 sequence of Kasalath was only changed from T to C at position 114 of the base, but it did not change the nature of the amino acids. No mutation was found at position 316 of GW2 Kasasth and the expression was not terminated prematurely. The designed dCAPs were labeled with endonuclease ApoI TD70 was 21bp and 30bp, while the Kasalath band was 51bp. Of the 240 RILs, 82 were found to contain GW2TD70, and 158 were GW2Kasalath. There were significant differences in heavy phenotypes. GW2TD70 gene was not found in 8 indica rice restorer lines and 10 japonica cultivars in Jiangsu province. GW2TD70 gene has certain regulation on grain width, and GW2TD70 gene can be used to improve grain weight in rice breeding.