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| Cloning, Expression and Promoter Analysis of FaGI Gene from Festuca arundinacea |
| SHU Jian-Hong1, LUO Wei2, LU Xue-Ping4, MU Qiong1, WU Jia-Hai2,3,*, WANG Xiao-Li1,* |
1 Institute of Prataculture, Guizhou Academy of Agricultural Sciences, Guiyang 550006, China;
2 School of Animal Science/Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, Guizhou University, Guiyang 550025, China;
3 Institute of Fruit Research, Guizhou Academy of Agricultural Sciences, Guiyang 550006, China;
4 College of Science,Guizhou University,Guiyang 550006, China |
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Abstract GIGANTEA (GI) is a rhythm related gene, which plays an important role controlling of rhythm output and flowering regulation in plants.To explore the biological function of GIGANTEA (GI) gene in tall fescue (Festuca arundinacea), in this study, tall fescue was used as the experimental material, and the gene sequence and promoter sequence were amplified by using RACE (rapid amplification of cDNA ends) cloning and chromosome walking method, and analyzed by bioinformatics. Sequence analysis showed that the total length of FaGI gene (GenBank No. MZ540915) was 3 869 bp, and its ORF was 3 447 bp, encoding 1 149 amino acids. The promoter sequence was 2 371 bp and had multiple cis-acting elements such as CURECORECR, GT1CONSENSUS, LTRE1HVBLT49 and MYB recognition site.The phylogenetic tree showed that the protein encoded by the FaGI gene had a close evolutionary relationship with the homologous proteins of gramineous plants such as Festuca pratensis, Lolium perenne, Secale cereale, Triticum aestivum and other grasses.The subcellular location showed that it was located in the nucleus, indicated that the protein might play a role in the nucleus. Fluorescence quantitative analysis of expression patterns under different light treatments and different developmental stages showed that the expression of FaGI gene was regulated by photoperiod and circadian clock. The expression level of FaGI gene was different in different developmental stages. The expression level was the highest in the seedling stage, and the expression level decreases in reproductive growth stage. The p1300-FaGI overexpression vector was constructed to transform Arabidopsis thaliana, and it was found that the Arabidopsis overexpression FaGI could up-regulate the expression of AtCCA1 (CIRCADIAN CLOCK ASSOCIATED 1), AtTOC1 (TIMING OF CAB EXPRESSION 1), and down-regulate the expression of AtFT (FLOWERING LOCUS T) and AtCO (CONSTANS). This study provides a theoretical basis for the follow-up verification of the FaGI gene function of tall fescue.
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Received: 12 March 2021
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Corresponding Authors:
* lhcbqk@163.com; wangxiaolizhenyuan@126.com
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[1] 姜敏. 2016. 大豆GmGI基因对植物开花及抗逆的调控研究[D]. 硕士学位论文, 东北农业大学, 导师: 李文滨, pp.28.
(Jiang M.Functional analysis of soybean GmGI gene for flowering and abiotic stress regulation[D]. Thesis for M.S., Northeast Agricultural University, Suppervisor: Li W B, pp. 28)
[2] 李芳. 2012. 大豆FKF1和GI基因的克隆、表达模式及功能分析[D]. 博士学位论文, 中国农业科学院, 导师: 傅永福, pp.55-59.
(Li F, Molecular cloning, expression profiles and functional analysis of FKF1 and GI gene in soybean[D]. Thesis for Ph.D., Chinese Academy of Agricultural Sciencs, Suppervisor: Fu Y F, pp. 55-59)
[3] 刘杨. 2014. 甘蓝型油菜生物节律钟输出基因BnGI的克隆和表达分析[J].上海交通大学学报(农业科学版), 32(5): 5-11.
(Liu Y.2014. Cloning and expression analysis of circadian clock output gene BnGI in Brassica napus L.[J]. Journal of Shanghai Jiaotong University (Agricultural Science), 32(5): 5-11.)
[4] 罗维, 舒健虹, 刘晓霞, 等. 2020. 高羊茅FaRVE8基因的克隆、亚细胞定位及表达分析[J].草业学报, 29(07): 60-69.
(Luo W, Shu J H, Liu X X, et al.Cloning, subcellular localization and expression analysis of the RVE8 gene from Festuca arundinacea[J]. Acta Prataculturae Sinica. 29(07): 60-69.)
[5] 孙霞, 王秀峰, 郑成淑, 等. 2012. 菊花节律钟输出基因CmGI(GIGANTEA)的cDNA全长克隆、序列信息及定量表达分析[J]. 中国农业科学. 45(13): 2690-2703.
(Sun X, Wang X F, Zheng C S.et al.2012. The cDNA cloning and analysis of sequence information and quantitative express of Chrysanthemum rhythms clock output gene CmGI (GIGANTEA)[J]. Scientia Agricultura Sinica, 45(13): 2690-2703.)
[6] 王新涛. 2009. 玉米光周期敏感基因ZmGI的克隆和功能分析[D]. 硕士学位论文. 河南农业大学, 导师: 陈彦慧. pp. 37-38.
(Wang X T.2009. Cloning and characteriation of photoperiod sensitive gene ZmGI in maize[D].Thesis for M.S., Henan Agricultural University, Suppervisor: Chen Y H, pp. 37-38)
[7] 肖图舰, 马玉华, 袁启凤, 等. 2020. 火龙果节律钟输出基因HpGI的克隆与表达分析[J].热带作物学报, 41(07): 1298-1304.
(Xiao T J, Ma Y H, Yuan Q F, et al.2020. Molecular cloning and expression analysis of rhythms clock output gene HpGI from Hylocereus polyrhizus[J]. Chinese Journal of Tropical Crops. 41(07): 1298-1304.)
[8] 赵翔宇. 2005. 小麦花发育重要基因TaGI1与TaMADS1的分离与功能分析[D]. 博士学位论文, 山东农业大学, 导师: 张宪省, pp.96-100.
(Zhao X Y, Isolation of TaGI1 and TaMADS1 gene and functional analysis in Triticum aestivum[D]. Thesis for Ph.D., Shandong Agricultural University, Suppervisor: Zhang X S, pp. 96-100.)
[9] Dae Y H, Sangkyu P, Sungbeom L, et al.2019. GIGANTEA regulates the timing stabilization of CONSTANS by altering the interaction between FKF1 and ZEITLUPE[J]. Molecular and Cells. 42(10): 693-701.
[10] Dalchau N, Baek S J, Briggs H M, et al.2011. The circadian oscillator gene GIGANTEA mediates a long-term response of the Arabidopsis thaliana circadian clock to sucrose[J]. Proceedings of the National Academy of Sciences of the USA, 108(12): 5104-5109.
[11] Edwards J, Martin A P, Andriunas F, et al.2010. GIGANTEA is a component of a regulatory pathway determining wall in growth deposition in phloem parenchyma transfer cells of Arabidopsis thaliana[J]. Plant Journal, 63(4): 651-661.
[12] Fowler S G, Cook D.2005. Thomashow M F. Low temperature induction of Arabidopsis CBF1, 2, and 3 is gated by the circadian clock[J]. Plant Physiology, 137(3): 961-968.
[13] Günl M, Liew E F, David K, et al.2009. Analysis of a post translational steroid induction system for GIGANTEA in Arabidopsis[J]. BMC Plant Biology, 9: 141.
[14] Hayama R, Izawa T, Shimamoto K.2002. Isolation of rice genes possibly involved in the photoperiodic control of flowering by afluorescent differential display method[J]. Plant & Cell Physiology, 43(5): 494-504.
[15] Hong S Y, Lee S, Seo P J, et al.2010. Identification and molecular characterization of a Brachypodium distachyon GIGANTEA gene: Functional conservation in monocot and dicot plants[J]. Plant MolecularBiology, 72(4-5): 485-497.
[16] Hsu P Y, Harmer S L.2014. Wheels within wheels: The plant circadian system[J]. Trends Plant Science, 19(4): 240-249.
[17] Imaizumi T, Schultz T F, Harmon F G, et al.2005. FKF1 F-box protein mediates cyclic degradation of a repressor of CONSTANS in Arabidopsis[J]. Science, 309(5732): 293-297.
[18] Kim T S, Wang L, Kim Y J, et al.2020. Compensatory mutations in GI and ZTL may modulate temperature compensation in the circadian clock[J]. Plant Physiology,182(2): 1130-1141.
[19] Kim W Y, Fujiwara S, Suh S S, et al.2007. ZEITLUPE is a circadian photoreceptor stabilized by GIGANTEA in blue light[J]. Nature, 449(7160): 356-360.
[20] Lee C M, Li M W, Feke A, et al.2019. IGANTEA recruits the UBP12 and UBP13 deubiquitylases to regulate accumulation of the ZTL photoreceptor complex[J]. Nature Communication. 10(1): 3750
[21] Mizoguchi T, Wright L, Fujiwara S, et al.2005. Distinct roles of GIGANTEA in promoting flowering and regulating circadian rhythms in Arabidopsis[J]. Plant Cell, 17(8): 2255-2270.
[22] Nohales M A, Kay S A.2016. Molecular mechanisms at the core of the plant circadian oscillator[J]. Nature Struct & Molecular Biology, 23(12): 1061-1069.
[23] Nohales M A, Kay S A.2019. GIGANTEA gates gibberellin signaling through stabilization of the DELLA proteins in Arabidopsis[J]. Proceedings of the National Academy of Sciences of the USA. 116(43): 21893-21899.
[24] Park D H, Somers D E, Kim Y S, et al.1999. Control of circadian rhythms and photoperiodic flowering by the Arabidopsis GIGANTEA gene[J]. Science. 285(5433): 1579-1582.
[25] Sawa M, Nusinow D A, Kay S A, et al.2007. FKF1 and GIGANTEA complex formation is required for day-length measurement in Arabidopsis[J]. Science, 318(5848): 261-265.
[26] Suárez-López P, Wheatley K, Robson F, et al.2001. CONSTANS mediates between the circadian clock and the control of flowering in Arabidopsis[J]. Nature, 410(6832): 1116-1120.
[27] Yano M, Katayose Y, Ashikari M, et al.2000. Hd1, a major photoperiod sensitivity quantitative trait locus in rice, is closely related to the Arabidopsis flowering time gene CONSTANS[J]. The Plant Cell, 12(12): 2473-2483. |
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