巨桉EgrDREB2A基因结构及表达特性分析

doi:10.11707/j.1001-7488.20150210

Abstract

Abstract:

【Objective】 A gene, EgrDREB2A , was isolated from the mRNA suppression subtractive hybridization library of Eucalyptus grandis (Eucgr. G03094). Based on the analysis of structure, subcellular localization of EgrDREB2A protein and gene expression under different treatments of low temperature, ABA and salt, the roles of EgrDREB2A in the resistance to abiotic stresses of Eucalyptus grandis were discussed. 【Method】 SMART and MatInspector softwares were used to analyze the protein structure of EgrDREB2A and the cis-elements in promoter sequence of the gene. Phylogenetic tree of DREB proteins was constructed by MEGA software. Subcellular localization of EgrDREB2A was characterized with the method of introducing EgrDREB2A-GFP fused genes into onion epidermal cells via gene gun bombardment. And, gene expression analysis in different tissue, under treatments of low temperature, ABA, salt and circadian rhythm were carried out by semi-quantitative and quantitative RT-PCR method respectively. For the gene co-expression of EgrDREB2A under different time treatment at 4 ℃, WGCNA and Cytoscape softwares were used. 【Result】 EgrDREB2A was classed into DREB2 group because the protein it encodes containing one AP2 domain which including a YRG and a RAYD conserved regions. The phylogenetic tree based on homology comparison showed it belonging to subtypeⅠof DREB2 group. Several cis-elements related with plant stress response were found in the EgrDREB2A promoter sequence. Nuclear localization with DREB2 merged protein with GFP implied EgrDREB2A mainly located in the nucleus. qRT-PCR result of EgrDREB2A under 0 ℃, 2 ℃,4 ℃, 6 ℃ and 8 ℃ revealed it was induced. Time course (0.5, 2, 6, 12, 24, 48 h) treatments to E. grandis seedlings at 4 ℃ also increased its expression as the time delayed. The genes co-expressed with EgrDREB2A under different time treatment at 4 ℃ were mostly associated with plant response to abiotic stresses. EgrDREB2A expression increased at the first and then decreased under 100 μmol·L^-1 ABA treatment. However, under the salt stress (200 mmol·L^-1), it was inhibited firstly and then induced. Circadian rhythm also regulates the EgrDREB2A expression, and the transcription level of it was promoted under light and hampered in the darkness. 【Conclusion】 EgrDREB2A is a transcriptional factor which was classified into DREB2 group. The expression of it was induced by low temperature and ABA, salt treatments and circadian rhythm also changed its transcription level. Most of cis-elements found in the promoter of EgrDREB2A and genes co-expressed with it were all associated with plant stress response. All these results showed EgrDREB2A possibly played an important role in the process of resistance to aboitic stresses in E. grandis.

Key words: Eucalyptus grandis, EgrDREB2A, gene expression, abiotic stress, circadian rhythm

CLC Number:

S718.46

Wei Xiaoling, Cheng Longjun, Dou Jinqing, Xu Fenghua. The Structure and Expression Characteristics of EgrDREB2A Gene in Eucalyptus grandis[J]. Scientia Silvae Sinicae, 2015, 51(2): 80-89.

References

王京京, 童再康, 黄程前, 等. 2012. 巨桉EgrCBF 1和EgrCBF2 基因的克隆和胁迫响应表达分析. 林业科学, 48(10): 41-48.
(Wang J J, Tong Z K, Huang C Q, et al. 2012. Cloning of EgrCBF 1 and EgrCBF2 genes from Eucalyptus grandis and their expression under stresses. Scientia Silvae Sinicae, 48(10):41-48.)
王亚红, 刘缙, 王玉国. 2010. 高质量提取银杏种仁总 RNA 的改良方法. 中国农学通报, 26(15): 48-52.
(Wang Y H, Liu J, Wang Y G. 2010. An improved method RNA isolation from seeds of Ginkgo biloba L. Chinese Agricultural Science Bulletin, 26(15):48-52.)
Agarwal P K, Agarwal P, Reddy M K, et al. 2006. Role of DREB transcription factors in abiotic and biotic stress tolerance in plants. Plant Cell Rep, 25(12): 1263-1274.
Agarwal P, Agarwal P K, Nair S, et al. 2007. Stress-inducible DREB2A transcription factor from Pennisetum glaucum is a phosphoprotein and its phosphorylation negatively regulates its DNA-binding activity. Mol Genet Genomics, 277(2): 189-198.
Avonce N, Leyman B, Mascorro-Gallardo J O, et al. 2004. The Arabidopsis trehalose-6-P synthase AtTPS1 gene is a regulator of glucose, abscisic acid, and stress signaling. Plant Physiol, 136(3): 3649-3659.
Bieniawska Z, Espinoza C, Schlereth A, et al. 2008. Disruption of the Arabidopsis circadian clock is responsible for extensive variation in the cold-responsive transcriptome. Plant Physiol, 147(1): 263-279.
Chinnusamy V, Schumaker K, Zhu J K. 2004. Molecular genetic perspectives on cross-talk and specificity in abiotic stress signalling in plants. Journal of experimental botany, 55(395): 225-236.
Dubouzet J G, Sakuma Y, Ito Y, et al. 2003. OsDREB genes in rice, Oryza sativa L., encode transcription activators that function in drought-, high-salt- and cold-responsive gene expression. Plant J, 33(4): 751-763.
Egawa C, Kobayashi F, Ishibashi M, et al. 2006. Differential regulation of transcript accumulation and alternative splicing of a DREB2 homolog under abiotic stress conditions in common wheat. Genes Genet Syst, 81(2): 77-91.
Fowler S G, Cook D, Thomashow M F. 2005. Low temperature induction of Arabidopsis CBF1, 2, and 3 is gated by the circadian clock. Plant Physiol, 137(3): 961-968.
Herrera-Rodriguez M B, Perez-Vicente R, Maldonado J M. 2007. Expression of asparagine synthetase genes in sunflower (Helianthus annuus) under various environmental stresses. Plant Physiol Biochem, 45(1): 33-38.
Jaglo-Ottosen K R, Gilmour S J, Zarka D G, et al. 1998. Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance. Science, 280(5360): 104-106.
Kayal E W, Navarro M, Marque G, et al. 2006. Expression profile of CBF-like transcriptional factors genes from Eucalyptus in response to cold. Journal of Expression Botany, 57(10): 2455-2496.
Lim C J, Hwang J E, Chen H, et al. 2007. Over-expression of the Arabidopsis DRE/CRT-binding transcription factor DREB2C enhances thermotolerance. Biochem Biophys Res Commun, 362(2): 431-436.
Liu L, Zhu K, Yang Y, et al. 2008. Molecular cloning, expression profiling and trans-activation property studies of a DREB2-like gene from chrysanthemum (Dendranthema vestitum). J Plant Res, 121(2): 215-226.
Liu Q, Kasuga M, Sakuma Y, et al. 1998. Two transcription factors, DREB1 and DREB2, with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought- and low-temperature-responsive gene expression, respectively, in Arabidopsis. Plant Cell, 10(8): 1391-1406.
Matsukura S, Mizoi J, Yoshida T, et al. 2010. Comprehensive analysis of rice DREB 2 -type genes that encode transcription factors involved in the expression of abiotic stress-responsive genes. Mol Genet Genomics, 283(2): 185-196.
Mizoi J, Shinozaki K, Yamaguchi-Shinozaki K. 2012. AP2/ERF family transcription factors in plant abiotic stress responses. Biochim Biophys Acta, 1819(2): 86-96.
Nakashima K, Ito Y, Yamaguchi-Shinozaki K. 2009. Transcriptional regulatory networks in response to abiotic stresses in Arabidopsis and grasses. Plant Physiology, 149(1): 88-95.
Nakashima K, Takasaki H, Mizoi J, et al. 2012. NAC transcription factors in plant abiotic stress responses. Biochimica et Biophysica Acta (BBA)-Gene Regulatory Mechanisms, 1819(2): 97-103.
Novillo F, Medina J, Salinas J. 2007. Arabidopsis CBF1 and CBF3 have a different function than CBF2 in cold acclimation and define different gene classes in the CBF regulon. Proc Natl Acad Sci USA, 104(52): 21002-21007.
Okamuro J K, Caster B, Villarroel R, et al. 1997. The AP2 domain of APETALA2 defines a large new family of DNA binding proteins in Arabidopsis. Proc Natl Acad Sci USA, 94(13): 7076-7081.
Paul M J, Primavesi L F, Jhurreea D, et al. 2008. Trehalose metabolism and signaling. Annu Rev Plant Biol, 59: 417-441.
Puranik S, Sahu P P, Srivastava P S, et al. 2012. NAC proteins: regulation and role in stress tolerance. Trends in Plant Science, 17(6): 369-381.
Qin F, Sakuma Y, Tran L S, et al. 2008. Arabidopsis DREB2A-interacting proteins function as RING E3 ligases and negatively regulate plant drought stress-responsive gene expression. Plant Cell, 20(6): 1693-1707.
Sakuma Y, Liu Q, Dubouzet J G, et al. 2002. 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, 290(3): 998-1009.
Sakuma Y, Maruyama K, Osakabe Y, et al. 2006. Functional analysis of an Arabidopsis transcription factor, DREB2A, involved in drought-responsive gene expression. Plant Cell, 18(5): 1292-1309.
Shen Y G, Zhang W K, He S J, et al. 2003. An EREBP/AP2-type protein in Triticum aestivum was a DRE-binding transcription factor induced by cold, dehydration and ABA stress. Theor Appl Genet, 106(5): 923-930.
Stockinger E J, Gilmour S J, Thomashow M F. 1997. Arabidopsis thaliana CBF1 encodes an AP2 domain-containing transcriptional activator that binds to the C-repeat/DRE, a cis-acting DNA regulatory element that stimulates transcription in response to low temperature and water deficit. Proc Natl Acad Sci USA, 94(3): 1035-1040.
Usadel B, Blasing O E, Gibon Y, et al. 2008. Multilevel genomic analysis of the response of transcripts, enzyme activities and metabolites in Arabidopsis rosettes to a progressive decrease of temperature in the non-freezing range. Plant Cell Environ, 31(4): 518-547.
Wang S, Wei X L, Cheng L J, et al. 2014. Identification of a C2H2-type zinc finger gene family from Eucalyptus grandis and its response to various abiotic stresses. Biologia Plantarum, 58(2): 385-390.
Wang H, Liu D, Sun J, et al. 2005. Asparagine synthetase gene TaASN 1 from wheat is up-regulated by salt stress, osmotic stress and ABA. J Plant Physiol, 162(1): 81-89.
Xu S P, Wei Z M. 1998. Introduction to method of microprojectile bombardment and its application. Plant Physiology Communications, 34(1):41-43.
Zhao J, Ren W, Zhi D, et al. 2007. Arabidopsis DREB1A/CBF3 bestowed transgenic tall fescue increased tolerance to drought stress. Plant Cell Rep, 26(9): 1521-1528.

[1]	Liu Daofeng, Wang Xia, Dai Yin, Yang Jianfeng, Ma Jing, Li Mingyang, Sui Shunzhao. Cloning and Function Analysis of CpTAF10 from Wintersweet (Chimonanthus praecox) [J]. Scientia Silvae Sinicae, 2019, 55(6): 176-183.
[2]	Lu Huijun, Li Ziyi, Liang Hanyu, Yue Yuanzhi, Zhou Tianchang, Yang Yuzhang, Wang Yucheng, Ji Xiaoyu. Expression and Stress Tolerance Analysis of NAC24 from Tamarix hispida [J]. Scientia Silvae Sinicae, 2019, 55(3): 54-63.
[3]	Fan Weijian, Xiang Weifang, Wang Jing, Bai Penghua, Pan Lina, Yang Yixin, Zhu Gengping, Li Min. Effects of Insecticides and Ultraviolet on the Expression of sHSP12.2 Gene in Chouioia cunea [J]. Scientia Silvae Sinicae, 2019, 55(2): 128-136.
[4]	Zhang Yang, Du Lin, Tang Xianfeng, Liu Huanhuan, Zhou Gongke, Chai Guohua. Identification and Expression Pattern Analyses of Populus BAG Genes [J]. Scientia Silvae Sinicae, 2019, 55(1): 138-145.
[5]	Cao Dandan, Wei Jianrong, Wang Weichao, Wang Liping, Zhao Zhengping, Liu Shuwen, Ji Baozhong. Homalota nanjingensis(Coleoptera: Staphylinidae: Aleocharinae) Under Bark of Dead Pine Trees Infected by Bursaphelenchus xylophilus [J]. Scientia Silvae Sinicae, 2019, 55(1): 157-162.
[6]	Wei Mingke, Yu Jinjian, Huang Xiaolong, Liu Qiongyao, Huang Huahong, Lin Erpei, Tong Zaikang. Cloning, Expression and Single Nucleotide Polymorphisms Analysis of NAC Transcription Factor Gene ClNAC1 in Cunninghamia lanceolata [J]. Scientia Silvae Sinicae, 2018, 54(9): 49-59.
[7]	Wang Juan, Chen Hui. Identification and Expression of Cold Shock Domain-containing Protein Gene in Dendroctonus armandi [J]. Scientia Silvae Sinicae, 2018, 54(4): 58-66.
[8]	Zhang Hehua, Liu Jiaxin, Luo Chaobing, Zhang Lingyun. Cloning and Analysis of a Transcription Factor PwERF8 and the Promoter Sequences in Picea wilsonii [J]. Scientia Silvae Sinicae, 2018, 54(3): 48-60.
[9]	Ma Jing, Li Zheng, Chen Xinli, Zhang Shengyan, Sui Shunzhao, Li Mingyang. Cloning and Activity Analysis of CpEXP1 Gene Promoter from Chimonanthus praecox [J]. Scientia Silvae Sinicae, 2018, 54(3): 61-72.
[10]	Li Jianbo, Jia Huixia, Zhang Jin, Liu Bobin, Hu Jianjun, Wang Lijuan, Lu Mengzhu. Effect of Overexpression of Populus tomentosa WUSCHEL-related homeobox 4 (PtoWOX4a) on the Secondary Growth of Poplar [J]. Scientia Silvae Sinicae, 2018, 54(2): 52-59.
[11]	Liu Changying, Zhang Meng, Wei Congjin, Xu Yazhen, Cao Boning, Zheng Sha, Fan Wei, Xiang Zhonghuai, Zhao Aichun. Resistance of Arabidopsis thaliana Transformed with Mulberry MAPK5 under Stress Conditions [J]. Scientia Silvae Sinicae, 2017, 53(9): 35-44.
[12]	Sun Huayu, Li Lichao, Zhao Hansheng, Yang Yihong, Wang Sining, Gao Zhimin. Molecular Characteristics and SSR Marker Development of Two NAC Genes from Daemonorops jenkinsiana [J]. Scientia Silvae Sinicae, 2017, 53(8): 132-140.
[13]	Tang Feifei, Zhao Yulin, Wang Peilong, Feng Deming, Song Yi, Gao Caiqiu. Cloning and Stress Tolerance Analysis of ThP5CR from Tamarix hispida [J]. Scientia Silvae Sinicae, 2017, 53(7): 1-9.
[14]	Li Nan, Zheng Yongqi, Ding Hongmei, Liu Xinhong, Sheng Weitong, Jiang Bo, Li Haibo. Analysis on Differential Expression of Cold Resistance Related Genes of Casuarina equisetifolia under Low Temperature Stress [J]. Scientia Silvae Sinicae, 2017, 53(7): 62-71.
[15]	You Hanli, Yuan Yihang, Li Changjiang, Zhang Lingyun. Cloning and Expression Analysis of MYB Homologous Gene PwMYB20 from Picea wilsonii [J]. Scientia Silvae Sinicae, 2017, 53(5): 23-32.

The Structure and Expression Characteristics of EgrDREB2A Gene in Eucalyptus grandis

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments