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Acta Agron Sin ›› 2013, Vol. 39 ›› Issue (08): 1352-1359.doi: 10.3724/SP.J.1006.2013.01352

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning and Expression Analysis of SbDREB2 Gene from Sorghum bicolor

XIE Deng-Lei1,CUI Jiang-Hui2,CHANG Jin-Hua1,*   

  1. 1 College of Agronomy, Agricultural University of Hebei, Baoding 071000, China; 2 College of Resources and Environment Science, Agricultural University of Hebei, Baoding 071000, China
  • Received:2013-02-04 Revised:2013-04-22 Online:2013-08-12 Published:2013-05-20
  • Contact: 常金华, E-mail: changjinhua@hebau.edu.cn
  • About author:常金华, E-mail: changjinhua@hebau.edu.cn

Abstract:

DREBs play important roles in regulating the expression of downstream genes in response to a variety of abiotic stresses. In this paper, a DREB-like gene, named SbDREB2, was assembled by searching sorghum EST and genome databases. The SbDREB2 gene was cloned from salinity-stressed sorghum seedling by RT-PCR. SbDREB2 contains a 789 bp complete open reading frame (ORF) which encodes a peptide of 262 amino acids. The predicted molecular weight and isoelctric point of SbDREB2 are 28.64 kD and 5.52, respectively. There is a 740 bp intron in the DNA sequence of SbDREB2. The amino acids analysis indicated that the predicted protein sequence contained a typical AP2 DNA-binding domain in the 82–145 regions. Multiple sequences alignment revealed that SbDREB2 shared 84% and 69% sequence similarities with Zea mays DREB2A and Oryza sativa DREB1, respectively. Prokaryotic expression vector pET28a-SbDREB2 was established and transformed BL21 (DE3) into E. coli after IPTG induction, showing a successful gene expression. The expression pattern analysis carried out by quantitative real-time PCR indicated that SbDREB2 was constitutively expressed in various tissues of sorghum, and was strongly up-regulated under high salinity, drought and exogenous application of abscisic acid (ABA). However, the expression of SbDREB2 was not affected by low temperature.

Key words: Sorghum, SbDREB2, Prokaryotic expression, Abiotic stress, Quantitative real-time PCR

[1]Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi-Shinozaki K, Shinozaki K. 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, 1998, 10: 1391–1406



[2]Shinozaki K, Yamaguchi-Shinozaki K, Seki M. Regulatory network of gene expression in the drought and cold stress responses. Curr Opin Plant Biol, 2003, 6: 410–417



[3]Agarwal P K, Jha B. Transcription factors in plants and ABA dependent and independent abiotic stress signaling. Biol Plant, 2010, 54: 201–212



[4]Liu Q, Zhao N M, Yamaguchi-Shinozaki K, Shinozaki K. Regulatory role of DREB transcription factors in plant drought, salt and cold tolerance. Chin Sci Bull, 2000, 45: 970–975



[5]Kizis D, Lumbreras V, Pages M. Role of AP2/EREBP transcription factors in gene regulation during abiotic stress. FEBS Lett, 2001, 498: 187–189



[6]Haake V, Cook D, Riechmann J L, Pineda O, Thomashow M F, Zhang J Z. Transcription factor CBF4 is a regulator of drought adaptation in Arabidopsis. Plant Physiol, 2002, 130: 639–648



[7]Li X P, Tian A G, Luo G Z, Gong Z Z, Zhang J S, Chen S Y. Soybean DRE-binding transcription factors that are responsive to abiotic stresses. Theor Appl Genet, 2005, 110: 1355–1362



[8]Jaglo K R, Kleff S, Amundsen K L, Zhang X, Haake V, Zhang J Z, Deits T, Thomashow M F. Components of the Arabidopsis C-repeat/dehydration-responsive element binding factor cold-response pathway are conserved in Brassica napus and other plant species. Plant Physiol, 2001, 127: 910–917



[9]Shen Y G, Zhang W K, He S J, Zhang J S, Liu Q, Chen S Y. An EREBP/AP2-type protein in Triticum aestivum was a DRE-binding transcription factor induced by cold, dehydration and ABA stress. Theor Appl Genet, 2003, 106: 923–930



[10]Kume S, Kobayashi F, Ishibashi M, Ohno R, Nakamura C, Takumi S. Differential and coordinated expression of Cbf and Cor/Lea genes during long-term cold acclimation in two wheat cultivars showing distinct levels of freezing tolerance. Genes Gene Systems, 2005, 80: 185–197



[11]Gao S Q, Chen M, Xia L Q, Xiu H J, Xu Z S, Li L C, Zhao C P, Chen X G, Ma Y Z. A cotton (Gossypium hirsutum) DRE-binding transcription factor gene, GhDREB, confers enhanced tolerance to drought, high salt, and freezing stresses in transgenic wheat. Plant Cell Rep, 2009, 28: 301–311



[12]QIN F, Sakuma Y, Li J, Liu Q, Li Y Q, Shinozaki K, Yamaguchi 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



[13]Hong J P, Kim W T. Isolation and functional characterization of the Ca-DREBLP1 gene encoding a dehydration-responsive element binding-factor-like protein 1 in hot pepper ( Capsicum annuum L. cv. Pukang). Planta, 2005, 220: 875–888



[14]Liu X-Y(刘宣雨), Wang Q-Y(王青云), Liu S-J(刘树君), Song S-Q(宋松泉). Research progress of gene transformation methods in sorghum. Chin Bull Bot (植物学报), 2011, 46(2): 216–223 (in Chinese with English abstract)



[15]Nawaz K, Talat A, Iqra, Hussain K, Majeed A. Induction of salt tolerance in two cultivars of sorghum (Sorghum bicolor L.) by exogenous application of Proline at seedling stage. World Appl Sci J, 2010, 10: 93–99



[16]Sun L(孙璐), Zhou Y-F(周宇飞), Wang C(汪澈), Xiao M-J(肖木辑), Tao Y(陶冶), Xu W-J(许文娟), Huang R-D(黄瑞冬). Screening and identification of sorghum cultivars for salinity tolerance during germination. Sci Agric Sin (中国农业科学), 2012, 45(9): 1714–1722 (in Chinese with English abstract)



[17]Gao J M, Xia B X, Yang H, Qu R G, Gui Z, Luo F, Pei Z Y, Sun S J. Comparative study on four methods for quick extraction of sorghum genomic DNA. Agric Sci Technol, 2011, 12: 686–687



[18]Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2–??CT method. Methods, 2001, 25: 402–408



[19]Wang S-X(王少峡), Wang Z-Y(王振英), Peng Y-K(彭永康). Dehydration responsive element binding (DREB) transcription activator and its function in plant tolerance to environmental stresses. Plant Physiol J (植物生理学通讯), 2004, 40(1): 7–13 (in Chinese)



[20]Xie X-Z(谢先芝), Wu N-H(吴乃虎). Intron in high plant. Chin Sci Bull (科学通报), 2002, 47(10): 731–737 (in Chinese with English abstract)



[21]Sakuma Y, Liu Q, Dubouzet J G, Abe H, Shinozaki K, Yamaguchi 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



[22]Liu L Q, Zhu K, Yang Y F, Wu J, Chen F D, Yu D Y. Molecular cloning, expression profiling and trans-activation property studies of a DREB2-like gene from chrysanthemum (Dendranthema vestitum ). J Plant Res, 2008, 121: 215–226



[23]Fu X-Y(付晓燕), Peng R-H(彭日荷), Zhang Z(章镇), Qiao Y-S(乔玉山), Zhou J(周军), Zhu B(朱波), Gao F(高峰), Tian Y-S(田永生), Zhao W(赵伟), Xiong A-S(熊爱生), Yao Q-H(姚泉洪). Cloning and expression of transcription factor gene MrDREBA6 from Malus micromalus. J Fruit Sci (果树学报), 2009, 26(6): 761–768 (in Chinese with English abstract)



[24]Sun X-B(孙晓波), Liu J-B(刘金兵), Yu G-H(余桂红), Zhang X(张旭), Zhang P(张鹏), Ma H-X(马鸿翔). Cloning and expression characterization of a SbDREB gene from Salicornia bigelovii Torr. J Plant Genet Resourc (植物遗传资源学报), 2012, 13(1): 111–117 (in Chinese with English abstract)

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