欢迎访问作物学报,今天是

作物学报 ›› 2013, Vol. 39 ›› Issue (02): 230-237.doi: 10.3724/SP.J.1006.2013.00230

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

白菜脱水应答转录因子BpDREB1基因的克隆及功能研究

刘晓颖,陈丽媛,张竞秋,李嘉玮,高越,王振英*   

  1. 天津师范大学生命科学学院 / 天津市细胞遗传与分子调控重点实验室,天津300387
  • 收稿日期:2012-05-10 修回日期:2012-10-09 出版日期:2013-02-12 网络出版日期:2012-11-14
  • 通讯作者: 王振英, E-mail: wzycell@yahoo.com.cn
  • 基金资助:

    本研究由国家自然科学基金项目(31071671), 天津市科委支撑项目(11ZCKFNC0070)和天津师范大学市级重点实验室开放研究基金(201003)资助。

Isolation and Functional Analysis of a New DREB Transcription Factor (BpDREB1) from Brassica pekinesis

LIU Xiao-Ying,CHEN Li-Yuan,ZHANG Jing-Qiu,LI Jia-Wei,GAO Yue,WANG Zhen-Ying*   

  1. Tianjin Key Laboratory of Cytogenetical and Molecular Regulation / College of Life Sciences, Tianjin Normal University, Tianjin 300387, China
  • Received:2012-05-10 Revised:2012-10-09 Published:2013-02-12 Published online:2012-11-14
  • Contact: 王振英, E-mail: wzycell@yahoo.com.cn

摘要:

DREB1/CBF类转录因子在植物抵抗外界胁迫上起重要作用,利用这些基因改良作物抗逆性具有重要意义。本研究在白菜中分离到一个DREB类转录因子基因BpDREB1 (EF219470)。该基因序列全长647 bp,推测编码蛋白含213个氨基酸,相对分子量为23 kD,理论等电点为5.11,与白菜中该类转录因子序列同源性为94%。进化树表明,BpDREB1属于DREB亚家族中A1亚族。基因的诱导表达模式分析显示,BpDREB1被低温强烈、迅速诱导表达,并对干旱胁迫也有一定程度的响应,但对高盐处理几乎没有响应。过表达BpDREB1的转基因拟南芥经低温诱导后,其体内可溶性糖及脯氨酸含量大幅度提高。以上结果显示BpDREB1转录因子基因具有家族成员基因结构的特征,在低温、干旱应答途径中起重要作用。

关键词: 白菜, BpDREB1, 低温, 干旱

Abstract:

DREB1/CBF transcription factors play an important role in plant stress tolerance, and one of important significance to gain stress-tolerant crops by transgenic technologies. In this study, a DREB-like gene, named BpDREB1 (accession No. EF219470), was cloned from Chinese cabbage. The BpDREB1 cDNA was 647 bp in length, and encoded protein of 213 amino acids with an predicted molecular weight of 23 kDand a isoelectric point of5.11, and shared 94% similarity with other DREB transcription factor from Chinese cabbage. On the basis of multiple sequence alignment and phylogenetic analysis, BpDREB1 was classified in A-1 group of the DREB family. The expression patterns analysis indicated that BpDREB1 was strongly up-regulated at low temperature, also responded to dehydrationHowever, the expression of BpDREB1 was not affected by high salinity. The expression pattern of BpDREB1 was the same as that of other DREB transcription factors in A-1 group. Overexpression of BpDREB1 greatlyincreased the contents of total soluable sugar andfree prolinein transgenic Arabidopsis plants, demonstrating that transgenic Arabidopsis induced the expression of soluable sugar and prolin related genes to enhance the tolerance to stress. These results suggest that BpDREB1 from Chinese cabbage has the typical characteristic of DREB transcription factors, and functions under the stress conditions including low temperature and drought.

Key words: Chinese cabbage, BpDREB1, Low temperature, Dehydration

[1]Xu Z S, Chen M, Li L C, Ma Y Z. Functions and application of the AP2/ERF transcription factor family in crop improvement. J Integr Plant Biol, 2011, 53: 570–585



[2]Yamaguchi-Shinozaki K, Shinozaki K. Transcriptional regulatory networks in cellular responses and tolerance to dehydration and cold stresses. Ann Rev Plant Physiol, 2006, 57: 781–803



[3]Vij S, Tyagi A K. Emerging trends in the functional genomics of the abiotic stress response in crop plants. Plant Biotechnol J, 2007, 5: 361–380



[4]Century K, Reuber T L, Ratcliffe OJ. Regulating the regulators: the future prospects for transcription-factor-based agricultural biotechnology products. Plant Physiol, 2008, 147: 20–29



[5]Yang S, Vanderbeld B, Wan J, Huang Y. Narrowing down the targets: towards successful genetic engineering of drought tolerant crops. Mol Plant, 2010, 3: 469–490



[6]Motoaki S, Mari N, Junko I, Tokihiko N, Miki F, Youko O, Asako K, Maiko N, Akiko E, Tetsuya S, Masakazu S, Kenji A, Teruaki T, Kazuko Y S, Piero C, Jun Kawai, Y H , Kazuo S. Monitoring the expression profiles of 7000 Arabidopsis genes under drought, cold, and high-salinity stresses using a full-length cDNA microarry. Plant J, 2002, 31: 279–292



[7]Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi-Shinozaki K, Shinozaki K. Two transcription factors, DREB1 and DREB2, with an AP2/EREBP DNA-binding domain separate two cellular signal transduction pathways in drought- and low-temperature responsive gene expression in Arabidopsis. Plant Cell, 1998, 10: 1391–1406



[8]Bartels D, Sunkar R. Drought and salt tolerance in plants. Critical Rev Plant Sci, 2005, 24: 23–58



[9]Vinocur B, Altman A. Recent advances in engineering plant tolerance to abiotic stress: achievements and limitations. Curr Opin Biotechnol, 2005, 16, 123–132



[10]Umezawa T, Fujita M, Fujita Y, Yamaguchi-Shinozaki K, Shinozaki K. Engineering drought tolerance in plants: discovering and tailoring genes to unlock the future. Curr Opin Biotechnol, 2006, 17: 113–122



[11]Thomashow M F, Gilmour S J, Stockinger E J, Jaglo-Ottosen K R, Zarka D G. Role of the Arabidopsis CBF transcriptional activators in cold acclimation. Physiol Plant 2001, 112: 171–175



[12]Agarwal P K, Agarwal P, Reddy M K, Sopory S K. Role of DREB transcription factors in abiotic and biotic stress tolerance in plants. Plant Cell Rep, 2006, 25: 1263–1274



[13]Kim J. Perception, transduction, and networks in cold signaling. J Plant Biol, 2007, 50: 139–147



[14]Gao J P, Chao D Y, Lin H X. Understanding abiotic stress tolerance mechanisms: recent studies on stress response in rice. J Integr Plant Biol, 2007, 49: 742–750



[15]Nakashima K, Shinwari Z K, Sakuma Y, Seki M, Miura S, Shinozak K, Yamaguchi-Shinozaki K. Organization and expression of two arabidopsis DREB2 genes encoding DRE binding proteins involved in dehydration- and high-salinity responsive gene expression. Plant Mol Biol, 2000, 42: 657–665



[16]Tian X H, Li X P, Zhou H L, Zhang J S, Gong Z Z, Chen S Y. OsDREB4 genes in rice encode AP2-containing proteins that bind specifically to the dehydration-responsive element. J Integr Plant Biol, 2005, 47: 467–476



[17]Sakuma Y, Maruyama K, Osakabe Y, Qin F, Seki M, Shinozaki K, Yamaguchi-Shinozakia K. Functional analysis of an Arabidopsis transcription factor, DREB2A, involved in drought-responsive gene expression. Plant Cell, 2006, 18: 1292–1309



[18]Qin Q L, Liu J G, Zhang Z, Peng R H, Xiong A S, Yao Q H, Chen J M. Isolation, optimization, and functional analysis of the cDNA encoding transcription factor OsDREB1B in Oryza sativa L. Mol Breed, 2007, 19: 329–340



[19]Gao M J, Allard G, Byass L, Flanagan A M, Singh J. Regulation and characterization of four CBF transcription factors from Brassica napus. Plant Mol Biol, 2002, 49: 459–471



[20]Qin F, Sakuma Y, Li J, Liu Q, Liu Y Q, Shinozaki K, Yamaguchi-Shinozaki K. Cloning and functional analysis of a novel DREB1. CBF transcription factor involved in cold-responsive gene expression in Zea mays L. Plant Cell Physiol, 2004, 45: 1042–1052



[21]Chen M, Xu Z S, Xia L Q, Li L C, Cheng X G, Dong J H, Wang Q Y, Ma Y Z. Cold-induced modulation and functional analyses of the DRE-binding transcription factor gene, GmDREB3, in soybean (Glycine max L.). J Exp Bot, 2009, 60: 121–135



[22]Oh S J, Kwon C W, Choi D W, Song S I, Kim J K. Expression of barley HvCBF4 enhances tolerance to abiotic stress in transgenic rice. Plant Biotechnol J, 2007, 5: 646–656



[23]Chen J Q, Meng X P, Zhang Y, Xia M, Wang X P. Over-expression of OsDREB genes lead to enhanced drought tolerance in rice. Biotechnol Lett, 2008, 30: 2191–2198



[24]Huang B, Jin L G, Liu J Y. Identification and characterization of the novel gene GhDBP2 encoding a DRE binding protein from cotton (Gossypium hirsutum). J Plant Physiol, 2008, 165: 214–223



[25]Clough S J, Bent A F. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J, 1998, 16: 735–743



[26]Yemm E W, Willis A J. The estimation of carbohydrates in plant extracts by anthrone. Biochemical J, 1954, 57: 508–514



[27]Zhang D Z, Wang P H, Zhao H X. Determination of the content of free proline in wheat leaves. Plant Physiol Commun, 1990, 4: 62–65



[28]Sakuma Y, Liu Q, Dubouzeta J G, Abea H, Shinozaki K, Yamaguchi-Shinozaki K. DNA-binding specificity of the ERF/AP2 domain of arabidopsis DREBs, transcription factors involved in dehydration- and cold-inducible gene expression. Biochem Biophys Res Commun, 2002, 290: 998–1009



[29]Wang P-R(王平荣), Deng X-J(邓晓建), Gao X-L(高晓玲), Chen J(陈静), Wan J(万佳), Jiang H(姜华), Xu Z-J(徐正君). Progress in the study on DREB transcription factor. Hereditas (遗传), 2006, 28: 369–374 (in Chinese with English abstract)



[30]Zhang M(张梅), Liu W(刘炜), Bi Y-P(毕玉平), Wang Z-Z(王自章). Isolation and identification of PNDREB1: a new DREB transcription factor from peanut (Arachis hypogaea L.). Acta Agron Sin (作物学报), 2009, 35: 1973–1980 (in Chinese with English abstract)



[31]Okamuro J K, CASTER B, Villarroel R, Montagu M V, Jofuku K D. The AP2 domain of APETALA2 defines a large new family of DNA binding proteins in Arabidopsis. Proc Natl Acad Sci USA, 1997, 94: 7076–7081



[32]Igarashi Y, Yoshiba Y, Sanada Y, Yamaguchi-Shinozaki K, Wada K, Shinozaki K. Characterization of the gene for deltal-pyrroline-5-carboxylate synthetase and correlation between the expression of the gene and salt tolerance in Oryza sativa L. Plant Mol Biol, 1997, 33: 857–865



[33]Zhang M(张梅), Liu W(刘炜), Bi Y-P(毕玉平). Dehydration-responsive element-binding (DREB) transcription factor in plants and its role during abiotic stresses. Hereditas (遗传), 2009, 31: 236–244 (in Chinese with English abstract)
[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] 徐苗苗, 邸太妹, 王洁, 吴叶蝶, 刘恩贝, 王玉春, 王新超, 王璐. 外源槲皮素增强茶树抗寒性的分子机制[J]. 作物学报, 2026, 52(5): 1418-1429.
[3] 杨飚, 杜帅康, 张继旺, 石瑛, 张丽莉. 马铃薯III类POD基因家族的全基因组鉴定及其表达谱分析[J]. 作物学报, 2026, 52(2): 405-420.
[4] 胡城祯, 高维东, 孔斌雪, 王建飞, 车卓, 杨德龙, 陈涛. 小麦TaAPC11基因家族鉴定及TaAPC11-5B参与干旱胁迫的生物学功能研究[J]. 作物学报, 2026, 52(1): 148-164.
[5] 王雅致, 杨飚, 季香林, 石瑛, 张丽莉. 二倍体马铃薯抗旱资源鉴定及抗旱基因初步筛选[J]. 作物学报, 2026, 52(1): 72-84.
[6] 孔娜, 刘涛, 刘文婷, 陈刚, 文利超, 邓智超, 郭梅, 李伟, 郭永峰. 烟草NtCEP7基因克隆及其编码小肽在苗期抗旱中的作用分析[J]. 作物学报, 2026, 52(1): 249-261.
[7] 刘海波, 张蕾, 王立琦, 石晓丽, 周文莹, 崔国贤, 佘玮. 苎麻BnGCL1基因响应干旱胁迫的功能研究[J]. 作物学报, 2026, 52(1): 14-27.
[8] 姬炫彤, 卞春松, 金黎平, 李森, 秦军红, 李广存. 不同耐旱型马铃薯根际微生物对干旱的响应[J]. 作物学报, 2026, 52(1): 165-177.
[9] 高源, 王宇琦, 姜佳宁, 赵健雄, 王雪贺缘, 王浩宇, 张芮嘉, 徐晶宇, 贺琳. 玉米低温响应基因ZmNTL1ZmNTL5的鉴定及功能分析[J]. 作物学报, 2025, 51(9): 2318-2329.
[10] 马娟娥, 姚有华, 姚晓华, 吴昆仑, 崔永梅. 青稞HvERF039基因的克隆及功能研究[J]. 作物学报, 2025, 51(9): 2341-2357.
[11] 何鹏旭, 姚立蓉, 陈远玲, 闫妍, 张宏, 汪军成, 李葆春, 杨轲, 司二静, 孟亚雄, 马小乐, 王化俊. 大麦干旱胁迫萌发生理及分子机理的差异性与相关性研究[J]. 作物学报, 2025, 51(9): 2412-2432.
[12] 张建鹏, 王国瑞, 别海, 叶飞宇, 马晨晨, 梁小菡, 鲁晓民, 尚霄丽, 曹丽茹. 转录因子ZmMYB153通过ABA信号调节气孔运动增强玉米苗期抗旱性[J]. 作物学报, 2025, 51(7): 1827-1837.
[13] 陆雯佳, 汪军成, 姚立蓉, 张宏, 司二静, 杨轲, 孟亚雄, 李葆春, 马小乐, 王化俊. 大麦PRX基因家族全基因组鉴定及其干旱胁迫下的表达分析[J]. 作物学报, 2025, 51(5): 1198-1214.
[14] 李乔, 叶杨春, 常旭虹, 王德梅, 王艳杰, 杨玉双, 马瑞琦, 赵广才, 蔡瑞国, 张敏, 刘希伟. 花后高温干旱逆境对冬小麦光合特性和产量的影响[J]. 作物学报, 2025, 51(4): 1077-1090.
[15] 王林, 陈晓雨, 张文梦龙, 汪思琦, 程冰云, 程靖秋, 潘锐, 张文英. 大麦HvMYB2分子特性及响应干旱胁迫的功能分析[J]. 作物学报, 2025, 51(4): 873-887.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!