欢迎访问作物学报，今天是
作物学报

作物学报 ›› 2010, Vol. 36 ›› Issue (4): 539-548.doi: 10.3724/SP.J.1006.2010.00539

• 作物遗传育种·种质资源·分子遗传学 •    下一篇

大豆GmCOL4基因的克隆与分析

张清哲1，马锦花1，陈新建1,*，傅永福2,*   

  1. 1河南农业大学农学院，河南郑州450002；2中国农业科学院作物科学研究所/农作物基因资源与遗传改良国家重大科学工程，北京100081
  • 收稿日期:2009-07-28 修回日期:2009-12-10 出版日期:2010-04-12 网络出版日期:2010-02-05
  • 通讯作者: 陈新建, E-mail: xinjian@371.net; 傅永福, E-mail: fuyf@caas.net.cn
  • 基金资助:

    本研究由“十一五”国家科技支撑计划重点项目（2007BAD59B02），国家转基因植物研究与产业化专项（2008ZX0809-001）和国家高技术研究发展计划（863计划）（2006AA10A111，2007AA10Z119）项目资助。

Cloning and Analysis of GmCIL4 Gene in Glycine max L.

ZHANG Qing-Zhe1,MA Jin-Hua1,CHEN Xin-Jian1,*,FU Yong-Fu2,*   

  1. 1 College of Agronomy, Henan Agricultural University, Zhengzhou 450002, China; 2 Institute of Crop Sciences, National Key Facility of Crop Gene Resource and Genetic Improvement, Chinese Academy of Agricultural Sciences, Beijing 10081, China
  • Received:2009-07-28 Revised:2009-12-10 Published:2010-04-12 Published online:2010-02-05
  • Contact: CHEN Xin-Jian, E-mail: xinjian@371.net;FU Yong-Fu, E-mail: fuyf@caas.net.cn

PDF

1685

被引次数

6

摘要：

CONSTANS (CO)是植物光周期开花途径中的关键基因之一。通过RT-PCR和生物信息学的方法，克隆了大豆GmCOL4基因并分析其结构特征，用实时荧光定量PCR (quantitative real-time RT-PCR, qRT-PCR)研究了其转录特点。结果表明，GmCOL4的4个外显子编码一个具有B-box和CCT保守结构域的CO-like蛋白，在序列上与拟南芥(Arabidopsis thaliana) COL9相似性最高，为64.3%。分析其转录特征发现，GmCOL4表达主要受生物节律的影响，受光的调节作用较弱。器官特异性表达分析发现，GmCOL4主要在大豆叶片中表达，表达模式与COL9相似。这为大豆中CO基因家族的功能研究提供了重要的依据。

关键词: CONSTANS, 生物信息学, 生物节律钟, 光周期, 大豆

Abstract:

The model plant Arabosopsis thaliana has been identified to have four major flowering pathways. Among them, the photoperiod pathway integrates the light signal including light/dark cycle, light spectrum, light intensity, and light duration, to mediate flower initiation. CONSTANS(CO) is a key gene in photoperiodic flowering pathway and acts between genes of the circadian clock and meristem identity. CO encodes a protein containing two zinc finger regions (B-box I and II) near the amino terminus and a CCT (CO, CO-Like, TOC1) domain near the carboxy terminus. The CO from Arabidopsis thaliana is one member of the family comprised of 17 members, which can be classed into three subgroups based on their characters of functional domains. To elucidate the function of CO in flowering in soybean, cloned one of CO-like gene, named as GmCOL4, from Glycine max L. Kennong 18. Bioinformatics analysis revealed that GmCOL4 encoded a protein embedded two conserved domains, B-box and CCT, and belonged to subgroup III. Phylogenetic analysis based on the critical amino acid sequences indicated that GmCOL4 was much close to COL9 with similarity of 64.3%. The expression profiles of GmCOL4 by quantitative real-time RT-PCR (qRT-PCR) showed a similar pattern to that of COL9. GmCOL4 was largely regulated by biological circadian, while the light appeared weak effect on it. And GmCOL4 expressed mainly in leaves and had its highest amount in anthesis. The results suggested that GmCOL4 is one of the important genes in the regulation of flowering and photoperiodic in soybean. It paves a way to study the function of CO family in soybean and its application in soybean molecular breeding.

Key words: CINSTANS, Bioinformatics, Circadian clock, Photoperiod, Glycine max L.

 


[1] G arner W W, Allard H A. Effect of the relative length of day and night and other factors of the environment on growth and reproduction in plants. J Agric Res, 1920, 18: 553-606

[2] Shaw D, Goldman B D. Gender differences in influence of prenatal photoperiods on postnatal pineal melatonin rhythms and serum prolactin and follicle-stimulating hormone in the Siberian hamster(Phodopus sungorus). Endocrinology, 1995, 136: 4237-4246

[3] Putterill J, Robson F, Lee K, Simon R, Coupland G. The CONSTANS gene of Arabidopsis promotes flowering and encodes a protein showing similarities to zinc finger transcription factors. Cell, 1995, 80: 847-857

[4] Suarez-Lopez P, Wheatley K, Robson F, Onouchi H, Valverde F, Coupland G. CONSTANS mediates between the circadian clock and the control of flowering in Arabidopsis. Nature, 2001, 410: 1116-1120

[5] Imaizumi T, Schultz T F, Harmon F G, Ho L A, Kay S A. FKF1 F-box protein mediates cyclic degradation of a repressor of CONSTANS in Arabidopsis. Science, 2005, 309: 293-297

[6] Sawa M, Nusinow D A, Kay S A, Imaizumi T. FKF1 and GIGANTEA complex formation is required for day-length mea- surement in Arabidopsis. Science, 2007, 318: 261-265

[7] Valverde F, Mouradov A, Soppe W, Ravenscroft D, Samach A, Coupland G. Photoreceptor regulation of CONSTANS protein in photoperiodic flowering. Science, 2004, 303: 1003-1006

[8] Kardailsky I, Shukla V K, Ahn J H, Dagenais N, Christensen S K, Nguyen J T, Chory J, Harrison M J, Weigel D. Activation tagging of the floral inducer FT. Science, 1999, 286: 1962-1965

[9] Kobayashi Y, Kaya H, Goto K, Iwabuchi M, Araki T. A pair of related genes with antagonistic roles in mediating flowering signals. Science, 1999, 286: 1960-1962

[10] Onouchi H, Igeno M I, Perilleux C, Graves K, Coupland G. Mutagenesis of plants overexpressing CONSTANS demonstrates novel interactions among Arabidopsis flowering-time genes. Plant Cell, 2000, 12: 885-900

[11] Samach A, Onouchi H, Gold S E, Ditta G S, Schwarz-Sommer Z, Yanofsky M F, Coupland G. Distinct roles of CONSTANS target genes in reproductive development of Arabidopsis. Science, 2000, 288: 1613-1616

[12] Borden K L. RING fingers and B-boxes: Zinc-binding protein-protein interaction domains. Biochem Cell Biol, 1998, 76: 351-358

[13] Strayer C, Oyama T, Schultz T F, Raman R, Somers D E, Mas P, Panda S, Kreps J A, Kay S A. Cloning of the Arabidopsis clock gene TOC1, an autoregulatory response regulator homolog. Science, 2000, 289: 768-771

[14] Torok M, Elkin L D. Two B or not two B? Overview of the rapidly expanding B-box family of proteins. Differentiation, 2000, 67: 63-71

[15] Robson F, Costa M M, Hepworth S R, Vizir I, Pineiro M, Reeves P H, Putterill J, Coupland G. Functional importance of conserved domains in the flowering-time gene CONSTANS demonstrated by analysis of mutant alleles and transgenic plants. Plant J, 2001, 28: 619-631

[16] Yano M, Katayose Y, Ashikari M, Yamanouchi U, Monna L, Fuse T, Baba T, Yamamoto K, Umehara Y, Nagamura Y, Sasaki T. Hd1, a major photoperiod sensitivity quantitative trait locus in rice, is closely related to the Arabidopsis flowering time gene CONSTANS. Plant Cell, 2000, 12: 2473-2484

[17] Nemoto Y, Kisaka M, Fuse T, Yano M, Ogihara Y. Characterization and functional analysis of three wheat genes with homology to the CONSTANS flowering time gene in transgenic rice. Plant J, 2003, 36: 82-93

[18] Serrano G, Herrera-Palau R, Romero J M, Serrano A, Coupland G, Valverde F. Chlamydomonas CONSTANS and the evolution of plant photoperiodic signaling. Curr Biol, 2009, 19: 359-368

[19] Griffiths S, Dunford R P, Coupland G, Laurie D A. The evolution of CONSTANS-like gene families in barley, rice, and Arabidopsis. Plant Physiol, 2003, 131: 1855-1867

[20] Robert L S, Robson F, Sharpe A, Lydiate D, Coupland G. Conserved structure and function of the Arabidopsis flowering time gene CONSTANS in Brassica napus. Plant Mol Biol, 1998, 37: 763-772

[21] Holefors A, Opseth L, Ree Rosnes A K, Ripel L, Snipen L, Fossdal C G, Olsen J E. Identification of PaCOL1 and PaCOL2, two CONSTANS-like genes showing decreased transcript levels preceding short day induced growth cessation in Norway spruce. Plant Physiol Biochem, 2009, 47: 105-115

[22] Hu R-B(胡瑞波). Molecular Cloning, Expression Profiles and Functional Analysis of FT/TFL1 Genes in Soybean (Glycine max). PhD Dissertation of Chinese Academy of Agricultural Sciences, 2009 (in Chinese with English Abstract)

[23] Chen Q J, Zhou H M, Chen J, Wang X C. Using a modified TA cloning method to create entry clones. Anal Biochem, 2006, 358: 120-125 Cheng X F, Wang Z Y. Overexpression of COL9, a CONSTANS- LIKE gene, delays flowering by reducing expression of CO and FT in Arabidopsis thaliana. Plant J, 2005, 43: 758-768
[1] 金昱何, 王雪菲, 徐张一娃, 缪怡宁, 蒋云杰, 伊莹, 缪德麟, 朱静仪, 钟一帆, 陈铭亨, 方芳, 刘鹏. 外源激素对低温胁迫下大豆叶片叶绿素荧光参数及抗氧化酶系统的影响[J]. 作物学报, 2026, 52(6): 1817-1829.
[2] 唐宽强, 李公允, 宋美毅, 赵雪, 常春玲. 大豆株高性状全基因组关联分析及预测模型构建[J]. 作物学报, 2026, 52(6): 1743-1756.
[3] 胡赵, 钱润, 谢丰璞, 应素平. 水稻SPX基因家族鉴定及响应磷处理的表达分析[J]. 作物学报, 2026, 52(6): 1902-1912.
[4] 姚术, 郭凯悦, 翟慧慧, 姚佳慧, 邓文琪, 闫玲, 黄驰, 高阳, 俞嫣然, 赵振邦, 李英慧, 王晓波, 李佳佳. 大豆苗期耐低铁综合评价及优异种质筛选[J]. 作物学报, 2026, 52(5): 1373-1387.
[5] 牛丽, 王勇胜, 王长杰, 张宏, 孟亚雄, 李葆春, 杨轲, 马小乐, 姚立蓉, 司二静, 王化俊, 汪军成. 大麦NAC基因家族鉴定分析及HvNAC38的耐盐功能验证[J]. 作物学报, 2026, 52(3): 688-707.
[6] 张晴, 杨昱, 郭茜, 岳霈尧, 殷丛丛, 牛景萍, 赵晋忠, 杜维俊, 岳爱琴. 大豆GmARA6a的克隆及响应盐胁迫的功能分析[J]. 作物学报, 2026, 52(2): 480-493.
[7] 王克晶, 李向华. 我国珍稀的大豆属多年生烟豆和短绒野大豆物种遗传资源濒危性评估分析[J]. 作物学报, 2025, 51(8): 2009-2019.
[8] 孟然, 李赵嘉, 冯薇, 陈悦, 刘路平, 杨春燕, 鲁雪林, 王秀萍. 大豆不同生育时期耐盐性综合评价及耐盐种质筛选[J]. 作物学报, 2025, 51(8): 1991-2008.
[9] 贺红利, 张雨涵, 杨静, 程云清, 赵杨, 李星诺, 司洪亮, 张兴政, 杨向东. 大豆e1-as基因突变体的创制及生理分析[J]. 作物学报, 2025, 51(8): 2228-2239.
[10] 胡蒙, 沙丹, 张晟瑞, 谷勇哲, 张世碧, 李静, 孙君明, 邱丽娟, 李斌. 大豆分枝数QTL定位及候选基因筛选[J]. 作物学报, 2025, 51(7): 1747-1756.
[11] 凤舞剑, 冼晓青, 张新钵, 曹丹, 强承魁. 基于转录组和AlphaFold对稻瘟菌经典效应蛋白和水稻受体的快速鉴定[J]. 作物学报, 2025, 51(6): 1480-1488.
[12] 王琼, 邹丹霞, 陈兴运, 张威, 张红梅, 刘晓庆, 贾倩茹, 魏利斌, 崔晓艳, 陈新, 王学军, 陈华涛. 大豆开花时间和成熟期性状全基因组关联分析与候选基因预测[J]. 作物学报, 2025, 51(6): 1558-1568.
[13] 殷丛丛, 李睿琦, 岳霈尧, 李晨, 牛景萍, 赵晋忠, 杜维俊, 岳爱琴. 基于闭合哑铃介导等温扩增可视化检测大豆花叶病毒SC15方法的建立及应用[J]. 作物学报, 2025, 51(5): 1248-1260.
[14] 张恒, 冯雅岚, 田文仲, 郭彬彬, 张均, 马超. 小麦TaSnRK基因家族鉴定及在局部根区干旱下的表达分析[J]. 作物学报, 2025, 51(3): 632-649.
[15] 杨芳萍, 郭莹, 田媛媛, 徐玉凤, 王兰兰, 白斌, 展宗冰, 张雪婷, 徐银萍, 刘金栋. 甘肃省小麦地方品种春化光周期基因效应及抗寒性评价[J]. 作物学报, 2025, 51(2): 370-382.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备15008789号-2