《植物生理学报》 2013, 49(8): 778-786

T-DNA插入水稻OsGUN4基因导致淡黄叶突变

陈盛杰, 刘巧泉*

扬州大学农学院教育部植物功能基因组学重点实验室, 江苏扬州225009

通信作者：刘巧泉；E-mail: qqliu@yzu.edu.cn；Tel: 0514-87996648

摘　要：

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–103 bp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。

关键词：水稻; 淡黄叶突变体; 叶绿素; OsGUN4基因; T-DNA标签

收稿：2013-06-28   修定：2013-07-21

资助：江苏省大学生实践创新训练计划项目(2012JSSPITP1409)

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

CHEN Sheng-Jie, LIU Qiao-Quan*

Key Laboratory of Plant Functional Genomics of the Ministry of Education, College of Agriculture, Yangzhou University, Yang-zhou, Jiangsu 225009, China

Corresponding author: LIU Qiao-Quan; E-mail: qqliu@yzu.edu.cn; Tel: 0514-87996648

Abstract:

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 in ygl-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.

Key words: Oryza sativa L.; yellow-green leaf mutant; chlorophyll; OsGUN4 gene; T-DNA tagging