T-DNA插入水稻OsGUN4基因导致淡黄叶突变

T-DNA Tagging of the OsGUN4 Gene Resulted in the Yellow-Green Leaf Mutation in Rice (Oryza sativa L.)

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。

In present study, a novel yellow-green leaf mutant ygl-11 was selected from the transgenic progeny of the indica rice cultivar ＇Longtefu B＇, and its genetic behavior as well as mutated gene was carefully studied. The leaf of the ygl-11 mutant is yellow during the whole growing periods. The mutation could be stably inherited according to the observation of several generations. The genetic analysis results indicated that the yellow-leaf mutation was controlled by a recessive gene and co-segregated with the integrated T-DNA in ygl-11 mutant. By using the inverse- and TAIL-PCR methods with minor modification, the flanking genomic sequence of the T-DNA insertion was cloned from the mutant. After alignment between the cloned sequences and rice genomic sequences, the T-DNA was identified to insert into the 5＇-untranslational region of the OsGUN4 gene on chromosome 11. The result of semi-quantitative RT-PCR revealed that there was no expression of the OsGUN4 gene inygl-11 mutant. Real-time RT-PCR analysis showed that the OsGUN4 gene highly expresses in green tissue of wild type rice, but very low in root and seed.