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从籼稻‘龙特甫B’来源的转基因后代中筛选获得一份淡黄叶突变体ygl-11,并对其进行了遗传分析和基因克隆研究。遗传分析表明,该淡黄叶突变性状受1对隐性核基因控制,并且与导入的T-DNA标签共分离。采用T-DNA标签的方法,从淡黄叶突变体植株中克隆了T-DNA插入位点的侧翼序列;序列分析表明T-DNA插入在水稻第11染色体OsGUN4基因5’非翻译区–103bp处。RT-PCR分析结果表明,突变体植株叶片中OsGUN4基因不表达;定量RT-PCR结果显示该基因在幼嫩的绿色叶片组织中表达量最高,在根和种子等组织中的表达极低。
A yellowish-leaf mutant, ygl-11, was screened from transgenic progeny of Longtefu B from indica rice. Genetic analysis and gene cloning were also carried out. Genetic analysis showed that the pale yellow leaf trait was controlled by one recessive nuclear gene and was co-segregated with the introduced T-DNA tag. The flanking sequence of T-DNA insertion site was cloned by T-DNA tagging. The sequence analysis showed that T-DNA was inserted at -103bp of 5 ’untranslated region of rice OsGUN4 gene . RT-PCR analysis showed that the OsGUN4 gene was not expressed in the leaves of the mutant plants. The quantitative RT-PCR results showed that the expression of OsGUN4 gene was highest in young green leaves and very low in roots and seeds.