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Molecular cloning, expression profiling and trans-activation property studies of a DREB2-like gene from chrysanthemum (Dendranthema vestitum)

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Abstract

Dehydration responsive element binding (DREB) proteins are important transcription factors in plant stress response and signal transduction. In this study, a DREB homolog gene, DvDREB2A, was isolated from chrysanthemum (Dendranthema vestitum) by reverse transcriptase-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. It contained an open reading frame (ORF) of 1,471 bp encoding 366 amino acid residues and was classified as a DREB2 subfamily member based on multiple sequence alignment. The predicted protein sequence contained a typical AP2/EREBP DNA-binding domain near the N-terminal region. In yeast one-hybrid analysis DvDREB2A protein was specifically bound to DRE elements (core sequence, A/GCCGAC) and activated the expression of the reporter HIS3 and LacZ. Transient expression experiment suggested that DvDREB2A protein was localized to the nucleus of onion epidermis cells. Quantitative real-time PCR (QRT-PCR) experiments showed that expression level of DvDREB2A was significantly affected by heat, low temperature, drought, abscisic acid (ABA) and high salinity treatments. These results indicated that the DvDREB2A gene is a new member of the DREB transcription factors, which may play an important role in providing tolerance to environmental stresses.

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References

  • Agarwal P, Agarwal PK, Nair S, Sopory SK, Reddy MK (2007) Stress-inducible DREB2A transcription factor from Pennisetum glaucum is a phosphorprotein and its phosphorylation negatively regulates its DNA-binding activity. Mol Genet Genomics 277:189–198

    Article  PubMed  CAS  Google Scholar 

  • Allen MD, Yamasaki M, Ohme-Takagi M, Tatenol M, Suzuki M (1998) A novel mode of DNA recognition by a β-sheet revealed by the solution structure of the GCC-box binding-domain in complex with DNA. EMBO J 17:5484–5496

    Article  PubMed  CAS  Google Scholar 

  • Baker SS, Wilhelm KS, Thomashow MF (1994) The 5′- region of Arabidopsis thaliana cor75a has cis-acting elements that confer cold-, drought- and ABA-regulated gene expression. Plant Mol Biol 24:701–713

    Article  PubMed  CAS  Google Scholar 

  • Baniwal SK, Bharti K, Chan KY, Fauth M, Ganguli A, Kotak S, Mishra SK, Nover L, Port M, Scharf KD et al (2004) Heat stress response in plants: a complex game with chaperones and more than twenty heat stress transcription factors. J Biosci 29:471–487

    Article  PubMed  CAS  Google Scholar 

  • Bray E, Bailey-Serresand J, Weretilnyk E (2000) Responses to abiotic stresses. In: Buchanan B, Gruissem W, Rockville JR (eds) Biochemistry and molecular biology of plants. American Society of Plant Biologists, Rockville, pp 1158–1203

    Google Scholar 

  • Cao ZF, Li J, Chen F, Li YQ, Zhou HM, Liu Q (2001) Effect of two conserved amino acid residues on DREB1A function. Biochemistry (Mosc) 66:623–627

    Article  CAS  Google Scholar 

  • Chen JQ, Dong Y, Wang YJ, Liu Q, Zhang JS, Chen SY (2003) An AP2/EREBP-type transcription-factor gene from rice is coldinducible and encodes a nuclear-localized protein. Theor Appl Genet 107:972–979

    Article  PubMed  CAS  Google Scholar 

  • Choi DW, Rodriguez EM, Close TJ (2002) Barley Cbf3 gene identification, expression pattern, and map location. Plant Physiol 129:1781–1787

    Article  PubMed  CAS  Google Scholar 

  • Díaz-Martín J, Almoguera C, Prieto-Dapena P, Espinosa JM, Jordano J (2005) Functional interaction between two transcription factors involved in the developmental regulation of a small heat stress protein gene promoter. Plant Physiol 139:1483–1494

    Article  PubMed  Google Scholar 

  • Dingwall C, Laskey RA (1991) Nuclear targeting sequences—a consensus. Trends Biochem Sci 16:478–481

    Article  PubMed  CAS  Google Scholar 

  • Dubouzet JG, Sakuma Y, Ito Y, Kasuga M, Dubouzet EG, Miura S, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) OsDREB genes in rice, Oryza sativa L., encode transcription activators that function in drought-, high salt and cold- responsive gene expression. Plant J 33:751–763

    Article  PubMed  CAS  Google Scholar 

  • Gilmour SJ, Zarka DG, Stockinger EJ, Salazar MP, Houghton JM, Thomashow MF (1998) Low temperature regulation of the Arabidopsis CBF family of AP2 transcriptional activators as an early step in cold-induced COR gene expression. Plant J 16:433–442

    Article  PubMed  CAS  Google Scholar 

  • Gilmour SJ, Sebolt AM, Salazar MP, Everard JD, Thomashow MF (2000) Overexpression of the Arabidopsis CBF3 transcriptional activator mimics multiple biochemical changes associated with cold acclimation. Plant Physiol 124:1854–1865

    Article  PubMed  CAS  Google Scholar 

  • Haake V, Cook D, Riechmann JL, Pineda O, Thomashow MF, Zhang JZ (2002) Transcription factor CBF4 is a regulator of drought adaptation in Arabidopsis. Plant Physiol 130:639–648

    Article  PubMed  CAS  Google Scholar 

  • Iwasaki T, Kiyosue T, Yamaguchi-Shinozaki K, Shinozaki K (1997) The dehydration-inducible Rd17 (Cor47) gene and its promoter region in Arabidopsis thaliana. Plant Physiol 115:1287–1289

    Article  Google Scholar 

  • Jaglo KR, Amundsen KL, Zhang X, Haake V, Zhang JZ, Deits T, Thomashow MF (2001) 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 127:910–917

    Article  PubMed  CAS  Google Scholar 

  • Jaglo-Ottosen KR, Gilmour SJ, Zarka DG, Schabenberger O, Thomashow MF (1998) Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance. Science 280:104–106

    Article  Google Scholar 

  • Jiang C, Iu B, Singh J (1996) Requirement of a CCGAC cis-acting element for cold induction of the BN115 gene from winter Brassica napus. Plant Mol Biol 30:679–684

    Article  PubMed  CAS  Google Scholar 

  • Kasuga M, Liu Q, Miura S, Yamaguchi-Shinozaki K (1999) Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor. Nat Biotechnol 17:287–292

    Article  PubMed  CAS  Google Scholar 

  • Kasuga M, Miura S, Shinozaki K, Yamaguchi-Shinozaki K (2004) A combination of the Arabidopsis DREB1A gene and stress-inducible rd29A promoter improved drought and low-temperature stress tolerance in tobacco by gene transfer. Plant Cell Physiol 45:346–350

    Article  PubMed  CAS  Google Scholar 

  • Kizis D, Pages M (2002) Maize DRE-binding proteins DBF1 and DBF2 are involved in rab17 regulation through the drought-responsive element in an ABA-dependent pathway. Plant J 30:679–689

    Article  PubMed  CAS  Google Scholar 

  • Kurkela S, Borg-Franck M (1992) Structure and expression of kin2, one of two cold- and ABA-induced genes of Arabidopsis thaliana. Plant Mol Biol 19:689–692

    Article  PubMed  CAS  Google Scholar 

  • Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi-Shinozaki Y, Shinozaki K (1998) Two transcription factors, DREB1 and DREB2, with an EREBP/AP2 binding domain separate two-temperature-responsive gene expression, respectively, in Arabidopsis. Plant Cell 10:1391–1406

    Article  PubMed  CAS  Google Scholar 

  • Magome H, Yamaguchi S, Hanada A, Kamiya Y, Oda K (2004) Dwarf and delayed-flowering 1, a novel Arabidopsis mutant deficient in gibberellin biosynthesis because of overexpression of a putative AP2 transcription factor. Plant J 37:720–729

    Article  PubMed  CAS  Google Scholar 

  • Nakashima K, Shinwari AK, Sakuma Y, Seki M, Miura S, Shinozaki K, Yamaguchi-Shinozaki K (2000) Organization and expression of two Arabidopsis DREB2 genes encoding DRE-binding proteins involved in dehydration-and high-salinity responsive gene expression. Plant Mol Biol 42:657–665

    Article  PubMed  CAS  Google Scholar 

  • Narusaka Y, Nakashima K, Shinwari ZK, Sakuma Y, Furihata T, Abe H, Narusaka M, Shinozaki K, Yamaguchi-Shinozaki K (2003) Interaction between two cis-acting elements, ABRE and DRE, in ABA-dependent expression of Arabidopsis rd29A gene in response to dehydration and high-salinity stresses. Plant J 34:137–148

    Article  PubMed  CAS  Google Scholar 

  • Nordin K, Heino P, Palva ET (1991) Separate signal pathways regulate the expression of low temperature-induced gene in Arabidopsis thaliana (L.) Heyn. Plant Mol Biol 16:1061–1071

    Article  PubMed  CAS  Google Scholar 

  • Nordin K, Vahala T, Palva ET (1993) Differential expression of two related low-temperature-induced genes in Arabidopsis thaliana (L.) Heyn. Plant Mol Biol 21:641–653

    Article  PubMed  CAS  Google Scholar 

  • Novillo F, Alonso JM, Ecker JR, Salinas J (2004) CBF2/DREB1C is a negative regulator of CBF1/DREB1B and CBF3/DREB1A expression and plays a central role in stress tolerance in Arabidopsis. Proc Natl Acad Sci USA 101:3985–3990

    Article  PubMed  CAS  Google Scholar 

  • Okamuro JK, Caster B, Villarroel R, Montagu MV, Jofuku KD (1997) The AP2 domain of APETALA2 defines a large new family of DNA binding-proteins in Arabidopsis. Proc Natl Acad Sci USA 94:7076–7081

    Article  PubMed  CAS  Google Scholar 

  • Ouellet F, Vazquez-Tello A, Sarhan F (1998) The wheat wcs120 promoter is cold-inducible in both monocotyledonous and dicotyledonous species. FEBS Lett 423:324–328

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Qin F, Kakimoto M, Sakuma Y, Maruyama K, Osakabe Y, Tran Lam-Son P, Shinozaki K, Yamaguchi-Shinozaki K (2007) Regulation and functional analysis of ZmDREB2A in response to drought and heat stress in Zea mays L. Plant J 50:54–69

    Article  PubMed  CAS  Google Scholar 

  • Sakuma Y, Liu Q, Dubouzet JG, Abe H, Shinozaki K, Yamaguchi-Shinozaki K (2002) DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs, transcription factors involved in dehydration- and cold-inducible gene expression. Biochem Biophy Res Commun 290:998–1009

    Article  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Sakuma Y, Maruyama K, Qin F, Osakabe Y, Shinozaki K, Yamaguchi-Shinozaki K (2006b) Dual function of an Arabidopsis transcription factor DREB2A in water-stress-responsive and heat-stress-responsive gene expression. Proc Natl Acad Sci USA 103:18822–18827

    Article  PubMed  CAS  Google Scholar 

  • Shen YG, Zhang WK, He SJ, Zhang JS, Liu Q, Chen SY (2003) An EREBP/AP2-type protein in Triticum aestivum was a DER-binding transcription factor induced by cold, dehydration and ABA stress. Theor Appl Genet 106:923–930

    PubMed  CAS  Google Scholar 

  • Shinozaki K, Yamaguchi-Shinozaki K (2000) Molecular responses to dehydration and low temperature: differences and cross-talk between two stress signaling pathways. Curr Opin Plant Biol 3:217–223

    PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Shinwari ZK, Nakashima K, Miura S, Kasuga M, Seki M, Yamaguchi-shinozaki K, Shinozaki K (1998) An Arabidopsis gene family encoding DRE/CRT binding proteins involved in low temperature-responsive gene expression. Biochem Biophys Res Commun 22:161–170

    Article  Google Scholar 

  • Stockinger EJ, Gilmour SJ, Thomashow MF (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 94:1035–1040

    Article  PubMed  CAS  Google Scholar 

  • Sun W, Van Montagu M, Verbruggen N (2002) Small heat shock proteins and stress tolerance in plants. Biochim Biophys Acta 1577:1–9

    PubMed  CAS  Google Scholar 

  • Thomashow MF (1999) Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Annu Rev Plant Physiol Plant Mol Biol 50:571–599

    Article  PubMed  CAS  Google Scholar 

  • Weising K, Atkinson RG, Gardner RC (1995) Genomic fingerprinting by microsatellite-primed PCR: a critical evaluation. PCR Methods Appl 4:249–255

    PubMed  CAS  Google Scholar 

  • Yamaguchi-Shinozaki K, Shinozaki K (1993) The plant hormone abscisic acid mediates the drought-induced expression but not the seed-specific expression of rd22, a gene responsive to dehydration-stress in Arabidopsis thaliana. Mol Gen Genet 238:17–25

    PubMed  CAS  Google Scholar 

  • Yamaguchi-Shinozaki K, Shinozaki K (1994) A novel cis-acting element in an Arabidopsis gene is involved in responsiveness to drought, low-temperature, or high-salt stress. Plant Cell 6:251–264

    Article  PubMed  CAS  Google Scholar 

  • Yamaguchi-Shinozaki K, Shinozaki K (2005) Organization of cis-acting regulatory elements in osmotic- and cold-stress-responsive promoters. Trends Plant Sci 10:88–94

    Article  PubMed  CAS  Google Scholar 

  • Zhu JK (2002) Salt and drought stress signal transduction in plants. Annu Rev Plant Biol 53:247–273

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We are greatly thankful to Prof. Qiang Liu for kindly providing the yeast one-hybrid system. Prof. Shou Yi Chen is thankful for important research advice. This work was supported by an award grant for outstanding scholars from the Ministry of Education of China.

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Authors and Affiliations

  1. National Center for Soybean Improvement, National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, 210095, People’s Republic of China

    Liqing Liu, Kai Zhu, Yanfang Yang, Jian Wu & Deyue Yu

  2. College of Horticulture, Nanjing Agricultural University, Nanjing, People’s Republic of China

    Fadi Chen

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  1. Liqing Liu
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  2. Kai Zhu
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  6. Deyue Yu
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Correspondence to Deyue Yu.

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Liu, L., Zhu, K., Yang, Y. et al. Molecular cloning, expression profiling and trans-activation property studies of a DREB2-like gene from chrysanthemum (Dendranthema vestitum). J Plant Res 121, 215–226 (2008). https://doi.org/10.1007/s10265-007-0140-x

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  • DOI: https://doi.org/10.1007/s10265-007-0140-x

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