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
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.
李超, 肖恩宗, 白庆, 徐枫, 冀曦悦, 李春娥, 王玉琪. 水稻OsCLF基因的功能分析及其对铝毒胁迫的响应[J]. 农业生物技术学报, 2022, 30(5): 837-846.
LI Chao, XIAO En-Zong, BAI Qing, XU Feng, JI Xi-Yue, LI Chun-E, WANG Yu-Qi. Functional Analysis of Gene OsCLF and Its Response to Aluminum Stress in Rice (Oryza sativa). 农业生物技术学报, 2022, 30(5): 837-846.
[1] 杨志敏, 汪瑾 . 2003. 植物耐铝的生物化学与分子机理[J]. 植物生理与分子生物学学报 , 29(5): 361-366. (Yang Z M, Wang J.2003. Biochemical and molecular mechanism for the aluminum tolerance in plants[J]. Journal of Plant Physiology and Molecular Biology, 29(5): 361-366.) [2] Avramova Z.2009. Evolution and pleiotropy of TRITHO RAX function in Arabidopsis[J]. International Journal of Developmental Biology, 53(2-3): 371-381. [3] Cao R, Zhang Y.2004. The functions of E(Z)/EZH2-mediated methylation of lysine 27 in histone H3[J]. Current Opin‐ ion in Genetics & Development, 14(2): 155-164. [4] Cazzonelli C, Cuttriss A, Cossetto S, et al.2009. Regulation of carotenoid composition and shoot branching in Arabi dopsis by a chromatin modifying histone methyltransferase, SDG8[J]. Plant Cell, 21(1):39-53. [5] Chang Y N, Zhu C, Jiang J, et al.2019. Epigenetic regulation in plant abiotic stress responses[J]. Journal of Integra tive Plant Biology, 62(5): 563-580. [6] Chen L T, Lou M, Wang Y Y, et al.2010. Involvement of Arabidopsis histone deacetylase HDA6 in ABA and salt stress response[J]. Journal of Experimental Botany, 61(12): 3345-3353. [7] Goodrich J, Puangsomlee P, Martin M, et al.1997. A Poly comb-group gene regulates homeotic gene expression in Arabidopsis[J]. Nature, 386(6620): 44-51. [8] Jiang D H, Wang Y Q, Wang Y Z, et al.2008. Repression of flowering locus c and flowering locus t by the Arabidop sis polycomb repressive complex 2 components[J]. PLOS ONE, 3(10): e3404. [9] Kim J M, Sasaki T, Ueda M, et al.2015. Chromatin changes in response to drought, salinity, heat, and cold stresses in plants[J]. Frontiers in Plant Science, 6: 114. [10] Kim J M, To T K, Ishida J, et al.2008. Alterations of lysine modifications on histone H3 N-tail under drought stress conditions in Arabidopsis thaliana[J]. Plant and Cell [11] Kochian L V, Hoekenga O A, Pineros M A.2004. How do crop plants tolerate acid soils? -Mechanisms of alumi num tolerance and phosphorous efficiency[J]. Annual Review of Plant Biology, 55: 459-493. [12] Kochian L V, Pineros M A, Liu J P, et al.2015. Plant adapta tion to acid soils: The molecular basis for crop alumi num resistance[J]. Annual Review of Plant Biology, 66, 571-598. [13] Kwon C S, Lee D, Choi G, et al.2009. Histone occupancy de pendent and -independent removal of H3K27 trimethylation at cold-responsive genes in Arabidopsis[J]. Plant Journal, 60(1): 112-121. [14] Liu J, Deng S L, Wang H, et al.2016. Curly leaf regulates gene sets coordinating seed size and lipid biosynthesis[J]. Plant Physiology, 171(1): 424-436. [15] Liu N, Fromm M, Avramova Z.2014a. H3K27me3 and H3K4me3 chromatin environment at super-induced dehydration stress memory genes of Arabidopsis thaliana[J]. Molecular Plant, 7(3): 502-513. [16] Liu X Y, Luo J L, Li T T, et al.2021. SDG711 is involved in rice seed development through regulation of starch me tabolism gene expression in coordination with other his tone modifications[J]. Rice, 14(1): 25. [17] Liu X Y, Zhou C, Zhao Y, et al.2014b. The rice enhancer of zeste [E(z)] genes SDG711 and SDG718 are respectively involved in long day and short day signaling to mediate the accurate photoperiod control of flowering time[J]. Frontiers in Plant Science, 5: 591. [18] Margueron R, Trojer P, Reinberg D.2005. The key to develop ment: Interpreting the histone code?[J]. Current Opinion in Genetics & Development, 15(2): 163-176. [19] Ng D W, Wang T, Chandrasekharan M B, et al.2007. Plant SET domain-containing proteins: Structure, function and regulation[J]. Biochimica et Biophysica Acta, 1769(5-6): 316-329. [20] Peterson C L, Laniel M A.2004. Histones and histone modifi cations[J]. Current Biology, 14(14): R546-551. [21] Schubert D, Primavesi L, Bishopp A R, et al.2006. Silencing by plant polycomb-group genes requires dispersed tri methylation of histone H3 at lysine 27[J]. EMBO Jour nal, 25(19): 4638-4649. [22] Song Y G, Ji D D, Li S, et al.2012. The dynamic changes of DNA methylation and histone modifications of salt re sponsive transcription factor genes in soybean[J]. PLOS ONE, 7(7): e41274. [23] Tsuji H, Saika H, Tsutsumi N, et al.2006. Dynamic and re versible changes in histone H3-Lys4 methylation and H3 acetylation occurring at submergence-inducible genes in rice[J]. Plant and Cell Physiology, 47(7): 995-1003. [24] Wang Y Q, Li R H, Li D M, et al.2017. NIP1; 2 is a plasma membrane-localized transporter mediating aluminum uptake, translocation, and tolerance in Arabidopsis[J]. Proceedings of the National Academy of Sciences of the USA, 114(19): 5047-5052. [25] Yuan L Y, Liu X C, Luo M, et al.2013. Involvement of his tone modifications in plant abiotic stress responses[J]. Journal of Integrative Plant Biology, 55(10): 892-901. [26] Zhou H Y, Liu Y H, Liang Y W, et al.2020. The function of histone lysine methylation related SET domain group proteins in plants[J]. Protein Science, 29(5): 1120-1137.
