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Isolation and characterization of rl(t), a gene that controls leaf rolling in rice
作者姓名:Lei Li  ;Xiang Xue  ;Zongxiang Chen  ;Yafang Zhang  ;Yuyin Ma  ;Cunhong Pan  ;Junkai Zhu  ;Xuebiao Pan  ;Shimin Zuo
作者单位:[1]Key Laboratory of Crop Genetics and Physiology of Jiangsu Province, College of Agriculture, Yangzhou University/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Yangzhou 225000, China; [2]Department of Biochemical Engineering, Yangzhou Polytechnic College, Yangzhou 225000, China; [3]Lixiahe Agricultural Research Institute of Jiangsu Province, Yangzhou 225000, China
基金项目:We thank Dr. Qiaoquan Liu for kindly provid- ing the binary vector. We also thank Dr. Liaoxun Lu for his helpful suggestions. This work was supported by the National Natural Sci- ence Foundation of China (31201180); the Natural Science Foundation of Jiangsu Province (BK2008223 and BK2012690); The "333" Project research Funging of Jiangsu Province (BRA2013125); The Innovation of Fund of the Jiangsu Academy of Agricultural Science and Technology (CX(12)1003-4) and A Project Funded by the Pri- ority Academic Program Development of Jiangsu Higher Education Institutions.
摘    要:Moderate leaf rolling is one of the most important morphological traits in rice breeding for plant ideotype. Previous studies have shown that the rl(t) gene has a high breeding potential for developing hybrid-rice varieties with an ideal ideotype, because it leads to an appropriate leaf rolling index (LRI) of about 30 % in the heterozygous state, and had a positive effect on grain yield. In this study, we isolated rl(t) and performed a preliminary investigation of its function in regulating leaf rolling in rice. DNA sequencing identified a single base change (G to T) in the finely mapped region (11 kb) containing rl(t), and this is located in 3′-untranslated region (3′-UTR) of the only predicted gene, Roc5 (Rice outermost cell-specific). The expression level of Roc5 is significantly higher in the rl(t) mutant than in the wild-type. Using RNAi and overexpression analysis, we found that the expression level of Roc5 correlated with LRI and leaf bulliform area, and wasalso associated with leaf abaxial or adaxial rolling. These results confirmed that Roc5 controls leaf rolling in a dosagedependent manner. Bioinformatics analysis revealed a conserved 17-nt sequence (called the GU-rich element) in the 3′-UTR of HD-GL2 (Homeodomain-Glabra2) family genes including Roc5. Based on the model of this element in regulating mRNA stability in mammals, we speculate that the single nucleotide change in this element accounts for the higher expression level of Roc5 in the rl(t) mutant compared to the wild-type, which ultimately leads to adaxial rolling of the leaf. This discovery further enhances our knowledge of the molecular mechanisms underlying leaf rolling in rice.

关 键 词:水稻育种  基因家族  RL  卷叶  分离  控制  特性  理想株型

Isolation and characterization of rl (t), a gene that controls leaf rolling in rice
Lei Li,;Xiang Xue,;Zongxiang Chen,;Yafang Zhang,;Yuyin Ma,;Cunhong Pan,;Junkai Zhu,;Xuebiao Pan,;Shimin Zuo.Isolation and characterization of rl (t), a gene that controls leaf rolling in rice[J].Chinese Science Bulletin,2014,59(25):3142-3152.
Authors:Lei Li  Xiang Xue  Zongxiang Chen  Yafang Zhang  Yuyin Ma  Cunhong Pan  Junkai Zhu  Xuebiao Pan  Shimin Zuo
Institution:1. Key Laboratory of Crop Genetics and Physiology of Jiangsu Province, College of Agriculture, Yangzhou University/Key Laboratory of Plant Functional Genomics of the Ministry of Education, Yangzhou, 225000, China
2. Department of Biochemical Engineering, Yangzhou Polytechnic College, Yangzhou, 225000, China
3. Lixiahe Agricultural Research Institute of Jiangsu Province, Yangzhou, 225000, China
Abstract:Moderate leaf rolling is one of the most important morphological traits in rice breeding for plant ideotype. Previous studies have shown that the rl (t) gene has a high breeding potential for developing hybrid-rice varieties with an ideal ideotype, because it leads to an appropriate leaf rolling index (LRI) of about 30 % in the heterozygous state, and had a positive effect on grain yield. In this study, we isolated rl (t) and performed a preliminary investigation of its function in regulating leaf rolling in rice. DNA sequencing identified a single base change (G to T) in the finely mapped region (11 kb) containing rl (t), and this is located in 3′-untranslated region (3′-UTR) of the only predicted gene, Roc5 (Rice outermost cell-specific). The expression level of Roc5 is significantly higher in the rl (t) mutant than in the wild-type. Using RNAi and overexpression analysis, we found that the expression level of Roc5 correlated with LRI and leaf bulliform area, and was also associated with leaf abaxial or adaxial rolling. These results confirmed that Roc5 controls leaf rolling in a dosage-dependent manner. Bioinformatics analysis revealed a conserved 17-nt sequence (called the GU-rich element) in the 3′-UTR of HD-GL2 (Homeodomain-Glabra2) family genes including Roc5. Based on the model of this element in regulating mRNA stability in mammals, we speculate that the single nucleotide change in this element accounts for the higher expression level of Roc5 in the rl (t) mutant compared to the wild-type, which ultimately leads to adaxial rolling of the leaf. This discovery further enhances our knowledge of the molecular mechanisms underlying leaf rolling in rice.
Keywords:rl(t)  HD-GL2  GU-rich element  Rolled leaf mutant  Rice
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