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CbCOR15, A Cold-Regulated Gene from Alpine Chorispora bungeana, Confers Cold Tolerance in Transgenic Tobacco

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Abstract

Rapid amplification of cDNA ends was performed to isolate cold-regulated CbCOR15 (EF208112) from Chorispora bungeana. This alpine species is distributed in subnival areas. Transcripts were detected in the leaves, but not the roots, of plants acclimated to cold temperatures. Expression was induced at high levels at both 4°C and −4°C. In comparing its deduced protein sequence to that of AtCOR15a (cold-regulated 15a in Arabidopsis thaliana), the N terminus had less homology than the C terminus while still containing a region analogous to the chloroplast-targeted signal peptide of AtCOR15a. We also introduced CbCOR15, with the CaMV 35S promoter, into tobacco. Second-generation (T1) plants had significantly increased tolerance to chilling, as determined by their electrolyte leakage, chlorophyll content, and relative water content. Further freezing-stress experiments showed that the tolerance of transgenic lines was significantly greater than that of the nontransgenics. Although the degree of chilling and freezing tolerance in the transgenic plants was not directly correlated with the accumulated levels of CbCOR15, we could conclude that this gene confers cold tolerance.

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References

  • Altschul SF, Madden TL, Schafer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  CAS  PubMed  Google Scholar 

  • An LZ, Liu YH, Feng HY, Feng GN, Chen GD (2000) Studies on characteristics of element contents of altifrigetic subnival vegetation at the source area of Urumqi River. Acta Bot Boreal-Occident Sin 20:80–86

    Google Scholar 

  • Artus NN, Uemura M, Steponkus PL, Gilmour SJ, Lin C, Thomashow MF (1996) Constitutive expression of the cold regulated Arabidopsis thaliana COR15a gene affects both chloroplast and protoplast freezing tolerance. Proc Natl Acad Sci USA 93:13404–13409

    Article  CAS  PubMed  Google Scholar 

  • Ayitu R, Dunyan T, Li JZ, Fang Y (1998) The relationship between the structure of vegetative organs in Chorispora bungeana and its environment. J Xinjiang Agric Univ 21:273–277

    Google Scholar 

  • 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 

  • Bannai H, Tamada Y, Maruyama O, Nakai K, Miyano S (2002) Extensive feature detection of N-terminal protein sorting signals. Bioinformatics 18:298–305

    Article  CAS  PubMed  Google Scholar 

  • Cattivelli L, Bartels D (1990) Molecular cloning and characterization of cold-regulated genes in barley. Plant Physiol 93:1504–1510

    Article  CAS  PubMed  Google Scholar 

  • Chauvin LP, Houde M, Sarhan F (1993) A leaf-specific gene stimulated by light during wheat acclimation to low temperature. Plant Mol Biol 23:255–265

    Article  CAS  PubMed  Google Scholar 

  • Dhanaraj AL, Slovin JP, Rowland LJ (2005) Isolation of a cDNA clone and characterization of expression of the highly abundant, cold acclimation-associated 14 kDa dehydrin of blueberry. Plant Sci 16:949–957

    Article  Google Scholar 

  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  • Fu XY, Chang JF, An LZ, Zhang MX, Xu SJ, Chen T, Liu YH, Xin H, Wang JH (2006) Association of the cold-hardiness of Chorispora bungeana with the distribution and accumulation of calcium in the cells and tissues. Environ Exp Bot 55:282–293

    Article  CAS  Google Scholar 

  • Gilmour SJ, Artus NN, Thomashow MF (1992) cDNA sequence analysis and expression of two cold-regulated genes of Arabidopsis thaliana. Plant Mol Biol 18:13–21

    Article  CAS  PubMed  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  CAS  PubMed  Google Scholar 

  • Guy CL (1990) Cold acclimation and freezing stress tolerance: role of protein metabolism. Annu Rev Plant Physiol Plant Mol Biol 41:187–223

    CAS  Google Scholar 

  • Hofgen R, Willmitzer L (1988) Storage of competent cells for Agrobacterium transformation. Nucleic Acids Res 16:9877

    Article  CAS  PubMed  Google Scholar 

  • Jaakola L, Pirtila AM, Halonen M, Hohtola A (2001) Isolation of high quality RNA from bilberry (Vaccinium myrtillus L.) fruit. Mol Biotechnol 19:203–210

    Article  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  CAS  PubMed  Google Scholar 

  • Lin C, Thomashow MF (1992) DNA sequence analysis of a complementary DNA for cold-regulated Arabidopsis COR15 and characterization of the COR15 polypeptide. Plant Physiol 115:171–180

    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 

  • Liu S, Wang X, Fan Z, Pang Y, Sun X, Wang X, Tang K (2004) Molecular cloning and characterization of a novel cold-regulated gene from Capsella bursa-pastoris. DNA Seq 15:262–268

    CAS  PubMed  Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, p 440

    Google Scholar 

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

    Article  CAS  Google Scholar 

  • Nakayama K, Okawa K, Kakizaki T, Honma T, Itoh H, Inaba T (2007) Arabidopsis Cor15am is a chloroplast stromal protein that has cryoprotective activity and forms oligomers. Plant Physiol 144:513–523

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Sakai A, Larcher W (1987) Frost survival of plants. Responses and adaptation to freezing stress. In: Billings WD, Golley F, Lange OL, Olson JS, Remmert H (eds) Ecological studies, vol 62. Springer, Berlin, p 252

    Google Scholar 

  • Shimamura C, Ohno R, Nakamura C, Takumi S (2006) Improvement of freezing tolerance in tobacco plants expressing a cold-responsive and chloroplast-targeting protein WCOR15 of wheat. J Plant Physiol 163:213–219

    Article  CAS  PubMed  Google Scholar 

  • Steponkus PL, Uemura M, Joseph RA, Gilmour SJ, Thomashow MF (1998) Mode of action of the COR15a gene on the freezing tolerance of Arabidopsis thaliana. Proc Natl Acad Sci U S A 95:14570–14575

    Article  CAS  PubMed  Google Scholar 

  • Takumi S, Koike A, Nakata M, Kume S, Ohno R, Nakamura C (2003) Cold-specific and light-stimulated expression of a wheat (Triticum aestivum L.) Cor gene Wcor15 encoding a chloroplast-targeted protein. J Exp Bot 54:2265–2274

    Article  CAS  PubMed  Google Scholar 

  • Thomashow MF (1990) Molecular genetics of cold acclimation in higher plants. In: Scandalios J (ed) Genomic responses to environmental stress. Academic, San Diego, pp 99–131

    Chapter  Google Scholar 

  • Thomashow MF (1993) Genes induced during cold acclimation in higher plants. In: Steponkus PL (ed) Advances in low-temperature biology, vol 2. JAI Press, London, pp 183–210

    Google Scholar 

  • Thomashow MF (1998) Role of cold-responsive genes in plant freezing tolerance. Plant Physiol 118:1–8

    Article  CAS  PubMed  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  CAS  PubMed  Google Scholar 

  • Thomashow MF (2001) So what’s new in the field of plant cold acclimation? Lots! Plant Physiol 125:89–93

    Article  CAS  PubMed  Google Scholar 

  • Tsvetanov S, Ohno R, Tsuda K, Takumi S, Mori N, Atanassov A, Nakamura C (2000) A cold-responsive wheat (Triticum aestivum L.) gene wcor14 identified in a winter-hardy cultivar `Mironovska 808'. Genes Genet Syst 75:49–57

    Article  CAS  PubMed  Google Scholar 

  • Walker K, Croteau R (2000) Taxol biosynthesis-molecular cloning of a benzoyl-CoA-taxane 2a-O-benzoyltransferase cDNA from Taxus and functional expression in Escherichia coli. Proc Natl Acad Sci USA 97:13591–13596

    Article  CAS  PubMed  Google Scholar 

  • Weatherley PE (1950) Studies in the water relations of cotton plants: 1. The field measurement of water deficit in leaves. New Phytol 49:81–97

    Article  Google Scholar 

  • White TC, Simmonds D, Donaldson P, Singh J (1994) Regulation of BN115, a low-temperature-responsive gene from winter Brassica napus. Plant Physiol 106:917–928

    Article  CAS  PubMed  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 

Download references

Acknowledgments

This study has been supported by the National Outstanding Youth Foundation of China (30625008), the National High Technology Research and Development Program (863 Program) (2007AA021401), and the National Basic Research Program of China (973 Program) (2007CB108902). We thank Dr. Philip Hall for his elaborate reviews.

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Authors and Affiliations

  1. Key Laboratory of Arid and Grassland Agroecology of Ministry of Education, School of Life Sciences, Lanzhou University, Lanzhou, 730000, China

    Jing Si, Hua Zhang, Ya-jie Liu & Li-zhe An

  2. Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou, 730000, China

    Jian-hui Wang

  3. College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou, 730000, China

    Li-jing Zhang

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  1. Jing Si
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  2. Jian-hui Wang
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Correspondence to Li-zhe An.

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Si, J., Wang, Jh., Zhang, Lj. et al. CbCOR15, A Cold-Regulated Gene from Alpine Chorispora bungeana, Confers Cold Tolerance in Transgenic Tobacco. J. Plant Biol. 52, 593–601 (2009). https://doi.org/10.1007/s12374-009-9077-z

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