Skip to main content
SpringerLink
Log in

Stress-inducible expression of GmDREB1 conferred salt tolerance in transgenic alfalfa

  • Research Note
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

In attempt to improve salt tolerance of alfalfa (Medicago sativa L.) plants, a soybean DREB orthologue, GmDREB1, was introduced into alfalfa plants under the control of Arabidopsis Rd29A promoter. Its incorporation and expression in transgenic plants were confirmed by DNA and RNA gel-blot analyses. The level of salt tolerance of transgenic lines was significantly higher than that of wild-type control plants as measured by ion leakage, chlorophyll fluorescence value, contents of free proline and total soluble sugars. Moreover, northern blot analysis revealed that the transcript level of Δ1-pyrroline-5-carboxylate synthase (P5CS) was up-regulated in the transgenic plants. These results suggested that the stress-inducible expression of GmDREB1 conferred salt tolerance in transgenic alfalfa plants. In addition, no visible phenotypic alternations were observed in the transgenic plants due to the use of stress-inducible Rd29A promoter. It was the first to describe transgenic expression of DREB genes in alfalfa and these transgenic lines with high salt-tolerance are of significance in the forage breeding.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Baker SS, Wilhelm KS, Thomashow MF (1994) The 5′-region of Arabidopsis thaliana cor15a has cis-acting elements that confer cold-, drought- and ABA-regulated gene expression. Plant Mol Biol 24:701–713

    Article  CAS  PubMed  Google Scholar 

  • Bates LE, Waldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–207

    Article  CAS  Google Scholar 

  • Bekki A, Trinchant JC, Rigaud J (1987) Nitrogen fixation (C2H2 reduction) by Medicago nodules and bacteroids under sodium chloride stress. Physiol Plant 71:61–67

    Article  CAS  Google Scholar 

  • Bray EA (1997) Plant response to water deficit. Trends Plant Sci 2:48–54

    Article  Google Scholar 

  • Chen M, Wang QY, Cheng XG, Xu ZS, Li LC, Ye XG, Xia LQ, Ma YZ (2007) GmDREB2, a soybean DRE-binding transcription factor, conferred drought and high-salt tolerance in transgenic plants. Biochem Biophys Res Commun 353:299–305

    Article  CAS  PubMed  Google Scholar 

  • Chen M, Xu ZS, Xia LQ, Li LC, Cheng XG, Dong JH, Wang QY, Ma YZ (2009) Cold-induced modulation and functional analyses of the DRE-binding transcription factor gene, GmDREB3, in soybean (Glycine max L.). J Exp Bot 60:121–135

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356

    Article  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  CAS  PubMed  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  CAS  PubMed  Google Scholar 

  • Ginzberg I, Stein H, Kapulnik Y, Szabados L, Strizhov N, Schell J, Koncz C, Zilberstein A (1998) Isolation and characterization of two different cDNAs of Δ1-pyrroline-5-carboxylate synthase in alfalfa, transcriptionally induced upon salt stress. Plant Mol Biol 38:755–764

    Article  CAS  PubMed  Google Scholar 

  • Hsieh TH, Lee JT, Charng YY, Chan MT (2002) Tomato plants ectopically expressing Arabidopsis CBF1 show enhanced resistance to water deficit stress. Plant Physiol 130:618–626

    Article  CAS  PubMed  Google Scholar 

  • Hughes MA, Dunn MA (1996) The molecular biology of plant acclimation to low temperature. J Exp Bot 47:291–305

    Article  CAS  Google Scholar 

  • Ito Y, Katsura K, Maruyama K, Taji T, Kobayashi M, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2006) Functional analysis of rice DREB1/CBF-type transcription factors involved in cold-responsive gene expression in transgenic rice. Plant Cell Physiol 47:141–153

    Article  CAS  PubMed  Google Scholar 

  • Jaglo KR, Kleff S, 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  CAS  PubMed  Google Scholar 

  • Kasuga M, Liu Q, Miura S, Yamaguchi-Shinozaki K, Shinozaki K (1999) Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor. Nat Biotechnol 17:287–291

    Article  CAS  PubMed  Google Scholar 

  • Kerepesi I, Galiba G (2000) Osmotic and salt stress induced alteration in soluble carbohydrate content in wheat seedlings. Crop Sci 40:482–487

    Article  CAS  Google Scholar 

  • Li XP, Tian AG, Luo GZ, Gong ZZ, Zhang JS, Chen SY (2005) Soybean DRE-binding transcription factors that are responsive to abiotic stress. Thero Appl Genet 110:1355–1362

    Article  CAS  Google Scholar 

  • Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi-Shinozaki K, Shinozaki K (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 10:1391–1406

    Article  CAS  PubMed  Google Scholar 

  • Lutts S, Kiner JM, Bouharmont J (1996) NaCl-induced senescence in leaves of rice (Oryza sativa L.) cultivars differing in salinity resistance. Ann Bot 78:389–398

    Article  CAS  Google Scholar 

  • Maruyama K, Sakuma Y, Kasuga M, Ito Y, Seki M, Goda H, Shimada Y, Yoshida S, Shinozaki K, Yamaguchi-Shinozaki K (2004) Identification of cold-inducible downstream genes of the Arabidopsis DREB1A/CBF3 transcriptional factor using two microarray system. Plant J 38:982–993

    Article  CAS  PubMed  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bio-assays with tobacco tissue culture. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Oberschall A, Deak M, Torok K, Sass L, Vass I, Kovacs I, Feher A, Dudits D, Horvath GV (2000) A novel aldose/aldehyde reductase protects transgenic plants against lipid peroxidation under chemical and drought stresses. Plant J 24:437–446

    Article  CAS  PubMed  Google Scholar 

  • Oh SJ, Song SI, Kim YS, Jang HJ, Kim SY, Kim M, Kim YK, Nahm BH, Kim JK (2005) Arabidopsis CBF3/DREB1A and ABF3 in transgenic rice increased tolerance to abiotic stress without stunting growth. Plant Physiol 138:341–351

    Article  CAS  PubMed  Google Scholar 

  • Pellegrineschi A, Reynolds M, Pacheco M, Brito RM, Almeraya R, Yamagunchi-Schinozaki K, Hosington D (2004) Stressed-induced expression in wheat of the Arabidopsis thaliana DREB1A gene delays water stress symptoms under greenhouse conditions. Genome 47:493–500

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Seki M, Narusaka M, Abe H, Kasuga M, Yamaguchi-Shinozaki K, Carninci P, Hayashizaki Y, Shinozaki K (2001) Monitoring the expression of pattern of 1300 Arabidopsis genes under drought and cold stresses by using a full-length cDNA microarray. Plant Cell 13:61–72

    Article  CAS  PubMed  Google Scholar 

  • Shen YG, Zhang WK, Yan DQ, Du BX, Zhang JS, Liu Q, Chen SY (2003) Characterization of a DRE-binding transcription factor from a halophyte Atriplex hortensis. Theor Appl Genet 107:155–161

    CAS  PubMed  Google Scholar 

  • Singh SK, Sharma HC, Goswami AM, Datta SP, Singh SP (2000) In vitro growth and leaf composition of grapevine cultivars as affected by sodium chloride. Biol Plant 43:283–286

    Article  CAS  Google Scholar 

  • Somers DA, Samac DA, Olhoft PM (2003) Recent advances in legume transformation. Plant Physiol 131:892–899

    Article  CAS  PubMed  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  CAS  PubMed  Google Scholar 

  • Uchimiya H, Murasshige T (1974) Evaluation of parameters in isolation of viable protoplasts from cultured tobacco cells. Plant Physiol 54:944–963

    Article  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  CAS  PubMed  Google Scholar 

  • Yoshiba Y, Kiyosue T, Nakashima K, Yamaguchi-Shinozaki K, Shinozaki K (1997) Regulation of levels of proline as an osmolyte in Plants under water stress. Plant Cell Physiol 38:1095–1102

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This research was supported by the National Key Project of Scientific and Technical Supporting Programs from Ministry of Science & Technology of China (2008BADB3B09) and the Programme of Introducing Talents of Discipline to Universities Project (B07017).

Author information

Authors and Affiliations

  1. Key Laboratory of Vegetation Ecology, School of Life Sciences, Northeast Normal University, 130024, Changchun, China

    Taicheng Jin, Qing Chang, Wangfeng Li, Dongxu Yin, Zhijian Li, Deli Wang, Bao Liu & Lixia Liu

  2. The School of Life Sciences, Jilin Normal University, 136000, Siping, China

    Taicheng Jin

Authors
  1. Taicheng Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qing Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wangfeng Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Dongxu Yin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Zhijian Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Deli Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Bao Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Lixia Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lixia Liu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jin, T., Chang, Q., Li, W. et al. Stress-inducible expression of GmDREB1 conferred salt tolerance in transgenic alfalfa. Plant Cell Tiss Organ Cult 100, 219–227 (2010). https://doi.org/10.1007/s11240-009-9628-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-009-9628-5

Keywords

Search

Navigation