Cloning, subcellular localization and expression analysis of the RVE8 gene from Festuca arundinacea

doi:10.11686/cyxb2019452

Abstract

Abstract: The circadian clock is a complex regulatory network that promotes plant growth in the ever-changing environment. REVEILLE8 (RVE8) is an important gene in the circadian clock. In order to explore the molecular mechanism, the FaRVE8 gene of leaves from Festuca arundinacea was cloned by RT-PCR and RACE. It was found that the full length of FaRVE8 was 1881 bp, the open reading frame was 1236 bp, encoding 411 amino acids, which belonged to a MYB-like factor. The phylogenetic analysis showed that it is closely related to Brachypodium distachyon, Aegilops tauschii, and Hordeum vulgare. Real-time fluorescence quantitative analysis of FaRVE8 expression in F. arundinacea leaves under different light treatments showed that FaRVE8 was expressed under different light treatments and showed obvious circadian rhythm. Subcellular localization indicated that FaRVE8 is localized in the nucleus, and FaRVE8 may play an important role in the nucleus. These above studies indicate that the FaRVE8 gene plays an important role in regulating biological rhythms, laying the foundation for further study of the function and molecular regulation mechanisms of the FaRVE8 gene.

Key words: Festuca arundinacea, FaRVE8, gene cloning, expression analysis

LUO Wei, SHU Jian-hong, LIU Xiao-xia, WANG Zi-yuan, MU Qiong, WANG Xiao-li, WU Jia-hai. Cloning, subcellular localization and expression analysis of the RVE8 gene from Festuca arundinacea[J]. Acta Prataculturae Sinica, 2020, 29(7): 60-69.

References

[1] Harmer, Stacey L. The circadian system in higher plants. Annual Review of Plant Biology, 2009, 60(1): 357-377.
[2] Michael T P, McClung C R. Enhancer trapping reveals widespread circadian clock transcriptional control in Arabidopsis. Plant Physiology, 2003, 132: 629-639.
[3] Covington M F, Maloof J N, Straume M, et al. Global transcriptome analysis reveals circadian regulation of key pathways in plant growth and development. Genome Biology, 2008, 9(8): https://doi.org/10.1186/gb-2008-9-8-r130.
[4] Akhtar R A, Reddy A B, Maywood E S, et al. Circadian cycling of the mouse liver transcriptome, as revealed by cDNA microarray, is driven by the suprachiasmatic nucleus. Current Biology, 2002, 12(7): 540-550.
[5] Dodd A N. Plant circadian clocks increase photosynthesis, growth, survival, and competitive advantage. Science, 2005, 309: 630-633.
[6] Goodspeed D, Chehab E W, Min-Venditti A, et al. From the cover: Arabidopsis synchronizes jasmonate-mediated defense with insect circadian behavior. Proceedings of the National Academy of Sciences of the United States of America, 2012, 109(12): 4674-4677.
[7] Green R M, Tingay S, Wang Z Y, et al. Circadian rhythms confer a higher level of fitness to Arabidopsis plants. Plant Physiology, 2002, 129: 576-584.
[8] Ruts T, Matsubara S, Wiese-Klinkenberg A, et al. Aberrant temporal growth pattern and morphology of root and shoot caused by a defective circadian clock in Arabidopsis thaliana. The Plant Journal, 2012, 72(1): 154-161.
[9] Yerushalmi S, Yakir E, Green R M. Circadian clocks and adaptation in Arabidopsis. Molecular Ecology, 2011, 20(6): 1155-1165.
[10] Hsu P Y, Harmer S L. Wheels within wheels: The plant circadian system. Trends in Plant Science, 2014, 19: 240-249.
[11] Alabadi D, Yanovsky M J, Más P, et al. Critical role for CCA1 and LHY in maintaining circadian rhythmicity in Arabidopsis. Current Biology, 2002, 12(9): 757-761.
[12] Schaffer R, Ramsay N, Samach A, et al. The late elongated hypocotyl mutation of Arabidopsis disrupts circadian rhythms and the photoperiodic control of flowering. Cell, 1998, 93: 1219-1229.
[13] Wang Z Y, Tobin E M. Constitutive expression of the CIRCADIAN CLOCK ASSOCIATED 1 (CCA1) gene disrupts circadian rhythms and suppresses its own expression. Cell, 1998, 93: 1207-1217.
[14] Schaffer R, Landgraf J, Accerbi M, et al. Microarray analysis of diurnal and circadian-regulated genes in Arabidopsis. The Plant Cell, 2001, 13(1): 113-123.
[15] Alabadi D. Reciprocal regulation between TOC1 and LHY/CCA1 within the Arabidopsis circadian clock. Science, 2001, 293: 880-883.
[16] Farinas B, Mas P. Functional implication of the MYB transcription factor RVE8/LCL5 in the circadian control of histone acetylation. The Plant Journal, 2011, 66(2): 318-329.
[17] Reetika R, Nozomu T, Yingshan H P, et al. REVEILLE8 and PSEUDO-REPONSE REGULATOR5 form a negative feedback loop within the Arabidopsis circadian clock. PLoS Genetics, 2011, 7(3): e1001350.
[18] Gong W, Hea K, Covington M F, et al. The development of protein microarrays and their applications in DNA-protein and protein-protein interaction analyses of Arabidopsis transcription factors. Molecular Plant, 2008, 1(1): 27-41.
[19] Green R M, Tobin E M. Loss of the circadian clock-associated protein 1 in Arabidopsis results in altered clock-regulated gene expression. Proceedings of the National Academy of Sciences of the United States of America, 1999, 96(7): 4176-4179.
[20] Feng Y, Xu Y, Chu W, et al. The spatiotemporal expression patterns of vernalization related genes in wheat. Molecular Plant Breeding, 2017, 15(10): 3886-3892.
冯岳, 徐尧, 褚蔚, 等. 小麦春化作用相关基因的时空表达特性研究. 分子植物育种, 2017, 15(10): 3886-3892.
[21] Wu T, Lan Z Q. Comparison and analysis of methods of extracting total RNA from petals of Camellia sasanqua. Chinese Agricultural Science Bulletin, 2013, 29(28): 129-133.
吴田, 蓝增全. 茶梅花瓣总RNA提取方法的比较和分析. 中国农学通报, 2013, 29(28): 129-133.
[22] Sparkes I A, Runions J, Kearns A, et al. Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. Nature Protocols, 2006, 1(4): 2019-2025.
[23] Feng P P, Chen P, Hong W J, et al. Research progress of MYB transcription factor family in Arabidopsis thaliana. Life Science Research, 2016, 20(6): 555-560.
冯盼盼, 陈鹏, 洪文杰, 等. 拟南芥MYB转录因子家族研究进展. 生命科学研究, 2016, 20(6): 555-560.
[24] Chen S K. Transcriptome analysis under heat stress and genome-wide analysis of MYB gene family of a wheat relative species, Brachypodium distachyon. Yangling: Northwest A&F University, 2018.
陈守坤. 小麦亲缘物种二穗短柄草热胁迫下转录组分析和MYB基因家族分析. 杨凌: 西北农林科技大学, 2018.
[25] Liu Y. Cloning and analysis of an MYB transcription factor involved in regulation anthocyanin production in apricot fruit. Chongqing: Southwest University, 2018.
刘羽. 调控杏果实花色素苷合成MYB转录因子的克隆及表达分析. 重庆: 西南大学, 2018.
[26] Zhang Z S, Wang T K, Qiu M D, et al. Cloning and transcriptional activation verification of rice gene OsZ314. Molecular Plant Breeding, (2019-11-27) [2019-12-27]. http://kns.cnki.net/kcms/detail/46.1068.S.20191127.0939.004.html.
张志槊, 王天抗, 邱牡丹, 等. 水稻OsZ314基因克隆及转录激活验证. 分子植物育种, (2019-11-27) [2019-12-27]. http://kns.cnki.net/kcms/detail/46.1068.S.20191127.0939.004.html.
[27] Gould P D, Locke J C W, Larue C, et al. The molecular basis of temperature compensation in the Arabidopsis circadian clock. The Plant Cell Online, 2006, 18(5): 1177-1187.
[28] Pérez-García P, Pablo, Ma Y, et al. Time-dependent sequestration of RVE8 by LNK proteins shapes the diurnal oscillation of anthocyanin biosynthesis. Proceedings of the National Academy of Sciences, 2015, 112(16): 5249-5253.
[29] Farinas B, Mas P. Histone acetylation and the circadian clock: A role for the MYB transcription factor RVE8/LCL5. Plant Signaling & Behavior, 2011, 6(4): 541-543.