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| Functional Analysis of Gene OsCLF and Its Response to Aluminum Stress in Rice (Oryza sativa) |
| LI Chao1,2, XIAO En-Zong1, BAI Qing1, XU Feng1, JI Xi-Yue1, LI Chun-E1, WANG Yu-Qi1,* |
1 School of Environmental Science and Engineering, Guangzhou University, Guangzhou 510006, China;
2 Shanghai Fuxing Senior High School, Shanghai 200434, China |
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Abstract Histone methylation plays an important role in regulating gene expression in eukaryotes. The curly leaf protein (CLF), which regulates leaf curling in Arabidopsis, is an important histone methyltransferase and involved in a variety of cell development processes. In this study, the functions of OsCLF were preliminarily studied in rice (Oryza sativa). The overexpression vector pCAMBIA1300-35S-OsCLF was constructed and identified by double restriction endonuclease digestion and sequencing, and then transformed into Arabidopsis thaliana clf mutant. Phenotypic and biochemistry function analysis revealed that OsCLF of rice had the histone H3K27 tri-methyltransferase function, which was similar to CLF of A. thaliana, and the phenotypes of clf mutant could be functional complemented to wild type's by OsCLF. Fluorescence quantitative PCR analysis results showed that the relative expression level of OsCLF was higher under long-day condition than that in short-day condition, and its expression was significantly induced by aluminum stress (P<0.01). After that, the RNA interference vector pYLRNA-OsCLF-RNAi was transferred into wild type rice. Phenotype analysis indicated that the plant height, flag leaf length and width, pollen viability and seed setting rate of OsCLF-RNAi transgenic plants were all significantly decreased, but the tiller number and chlorophyll content were remarkablely increased than those of wild type rice (P<0.01). In addition, the OsCLF RNAi transgenic rice lines were more tolerant to aluminum stress than wild type rice, which indicated that OsCLF negatively regulated aluminum tolerance related genes' expression. This study preliminarily revealed the biological processes of rice growth and development involved by OsCLF gene, and provides a reference basis for further study on the epigenetic mechanism of these processes.
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Received: 19 July 2021
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Corresponding Authors:
*yqwang@gzhu.edu.cn
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