大豆乙烯响应因子GmERF6的功能分析

生物技术通报 ›› 2013, Vol. 0 ›› Issue (12): 73-77.

大豆乙烯响应因子GmERF6的功能分析

翟莹¹ 赵艳¹ 杨晓杰¹ 孙天国¹ 张军² 姚雅男³

1.齐齐哈尔大学生命科学与农林学院遗传重点实验室，齐齐哈尔 161006
2.黑龙江省兽医科学研究所，齐齐哈尔 161006；

收稿日期:2013-07-12 出版日期:2013-12-20 发布日期:2013-12-20
作者简介:翟莹，女，博士，讲师，研究方向：大豆分子育种；E-mail：fairy39809079@126.com
基金资助:
齐齐哈尔大学青年教师科研启动支持计划项目（2012k-M20）

Functional Analysis of Ethylene-response Factor GmERF6 in Soybean

Zhai Ying¹, Zhao Yan¹, Yang Xiaojie¹ ,Sun Tianguo¹, Zhang Jun², Yao Yanan³

1. Key Laboratory of Genetics，College of Life Science and Agro-Forestry，Qiqihaer Univesity，Qiqihaer 161006
2.Heilongjiang Institute of Veterinary Science ，Qiqihaer 161006
3.Changchun Agriculture Expo Garden ，Changchun 130117

Received:2013-07-12 Published:2013-12-20 Online:2013-12-20

摘要/Abstract

摘要： GmERF6 是从大豆中克隆到的一个新的ERF转录抑制子基因，对其功能作了进一步的分析。亚细胞定位试验结果显示GmERF6蛋白定位于细胞核中；利用原核表达纯化GmERF6蛋白并进行凝胶阻滞分析（EMSA），结果显示，GmERF6蛋白在体外可以与GCC-box元件特异结合；对GmERF6转基因拟南芥干旱条件下的种子萌发率和植株表型进行观察，结果证实GmERF6提高了转基因拟南芥的抗旱性，表明该基因可作为培育抗旱材料的候选基因。

关键词: 大豆, GmERF6, 亚细胞定位, EMSA, 抗旱性

Abstract: GmERF6 is a new ERF transcription repressor isolated from soybean. The function of GmERF6 was further analyzed in this research. GmERF6 protein localized to the nucleus when transiently expressed in onion epidermal cells. Furthermore, EMSA showed GmERF6 protein binding to the GCC-box element in vitro . The seed germination rates of GmERF6 transgenic Arabidopsis were significantly higher than that of wild type Arabidopsis under drought stress. GmERF6 transgenic Arabidopsis grew much healthier than wild-type Arabidopsis by removing water for 18 d. These data indicated that GmERF6 enhanced drought tolerance in transgenic Arabidopsis . These results suggest GmERF6 is likely to be a useful tool for improving drought tolerance of plants.

Key words: Soybean, GmERF6, Subcellular localization, EMSA, Drought tolerance

翟莹, 赵艳, 杨晓杰, 孙天国, 张军, 姚雅男. 大豆乙烯响应因子GmERF6的功能分析[J]. 生物技术通报, 2013, 0(12): 73-77.

Zhai Ying, Zhao Yan, Yang Xiaojie ,Sun Tianguo, Zhang Jun, Yao Yanan. Functional Analysis of Ethylene-response Factor GmERF6 in Soybean[J]. Biotechnology Bulletin, 2013, 0(12): 73-77.

参考文献

[1] Xiong L, Ishitani M, Zhu JK. Interaction of osmotic stress, temperature and abscisic acid in the regulation of gene expression in Arabidopsis [J]. Plant Physiol, 1999, 119（1）：205-211.
[2] Magnani E, Sjolander K, Hake S. From endonucleases to transcription factors：evolution of the AP2 DNA binding domain in plants[J]. Plant Cell, 2004, 16（9）：2265-2277.
[3] Riechmann JL, Ratcliffe OJ. A genomic perspective on plant transcription factors[J]. Curr Opin Plant Biol, 2000, 3：423-434.
[4] Gutterson N, and Reuber TL. Regulation of disease resistance pathways by AP2/ERF transcription factors[J]. Curr Opin Plant Biol, 2004, 7（4）：465-471.
[5] Ohme-Takagi M, Shinshi H. Ethylene-inducible DNA binding proteins that interact with an ethylene-responsive element[J]. Plant Cell, 1995, 7（2）：173-182.
[6] Park JM, Park CJ, Lee SB, et al. Overexpression of the tobacco Tsi1 gene encoding an EREBP /AP2-type transcription factor enhances resistance against pathogen attack and osmotic stressing tobacco[J]. Plant Cell, 13（5）：2001, 1035-1046.
[7] Yang Z, Tian L, Latoszek-Green M, et al. Arabidopsis ERF4 is a transcriptional repressor capable of modulating ethylene and abscisic acid responses[J]. Plant Mol Biol, 2005, 58（4）：585-596.
[8] Zhang G, Chen M, Chen X, et al. Isolation and characterization of a novel EAR-motifcontaining gene GmERF4 from soybean（Glycine max L.）[J]. Mol Biol Rep, 2010, 37（2）：809-818.
[9] Zhai Y, Li JW, Li XW, et al. Isolation and characterization of a novel transcriptional repressor GmERF6 from soybean[J]. Biol Plantarum, 2013, 57（1）：26-32.
[10] 刘肖飞, 梁卫红.根癌农杆菌介导的GFP在洋葱表皮细胞定位研究[J]. 河南师范大学学报, 2009, 37（1）：123-125.
[11] 翟莹, 雷婷婷, 闫帆, 等. 大豆GmERF6 基因的原核表达及重组蛋白纯化[J]. 大豆科学, 2011, 30（6）：906-909.
[12] Liu Y, Zhao TJ, Liu JM, et al. The conserved Ala37 in the ERF/AP2 domain is essential for binding with the DRE element and the GCC box[J]. FEBS Lett, 2006, 580（5）：1303-1308.
[13] Yi SY, Kim JH, Joung YH, et al. The pepper transcription factor CaPF1 confers pathogen and freezing tolerance in Arabidopsis [J]. Plant Physiol, 2004, 136（1）：2862-2874.
[14] Zuo KJ, Qin J, Zhao JY, et al. Over-expression GbERF2 transcription factor in tobacco enhances brown spots disease resistance by activating expression of downstream genes[J]. Gene, 2007, 391（1-2）：80-90.
[15] Kazan K. Negative regulation of defence and stress genes by EAR-motif-containing repressors[J]. Trends Plant Sci, 2006, 11（3）：109-112.
[16] Ciftci-Yilmaz S, Morsy MR, Song L, et al. The EAR-motif of the Cys2/His2-type zinc finger protein Zat7 plays a key role in the defense response of Arabidopsis to salinity stress[J]. Journal Biological Chem, 2007, 282（12）：9260-9268.
[17] Ohta M, Matsui K, Hiratsu K, et al. Repression domains of class II ERF transcriptional repressors share an essential motif for active repression[J]. Plant Cell, 2001, 13（8）：1959-1968.
[18] Gu Y, Wildermuth M, Chakravarthy S, et al. Tomato transcription factors Pti4, Pti5, and Pti6 activate defense responses when expressed in Arabidopsis [J]. Plant Cell, 2002, 14：817-831.
[19] Eady CC, Reader J, Davis S, Dale T. Inheritance and expression of introduced DNA in transgenic onion plants（Allium cepa ）[J]. Ann Appl Biol, 2003, 142（2）：219-224.
[20] Eady CC, Davis S, Catanach A, et al. Agrobacterium tumefaciens- mediated transformation of leek（Allium porrum ）and garlic（Allium sativum ）[J]. Plant Cell Rep, 2005, 24（4）：209-215.
[21] Lee JH, Hong JP, et al. The ethylene-responsive factor like protein 1
（CaERFLP1）of hot pepper（Capsicum annuum L.）interacts in vitro with both GCC and DRE/CRT sequences with different binding affinities：Possible biological roles of CaERFLP1 in response to pathogen infection and high salinity conditions in transgenic tobacco plants[J]. Plant Mol Biol, 2004, 55（1）：61-81.
[22] Meng XP, Li FG, Liu CL, et al. Isolation and characterization of an ERF transcription factor gene from cotton（Gossypium barbadense L.）[J]. Plant Mol Biol Rep, 2010, 28（1）：176-183.
[23] Onate-Sanchez L, Anderson JP, et al. AtERF14, a member of the ERF family of transcription factors, plays a nonredundant role in plant defense[J]. Plant Physiol, 2007, 143（1）：400-409.
[24] Dai X, Xu Y, Ma Q, et al. Overexpression of an R1R2R3 MYB gene, OsMYB3R-2, inerease tolerance to freezing, drought, and salt stress in transgenic Arabidopsis [J]. Plant Physiol, 2007, 143（4）：1739-1751.