Molecular Breeding

, Volume 30, Issue 4, pp 1611–1620 | Cite as

Cold-induced modulation of CbICE53 gene activates endogenous genes to enhance acclimation in transgenic tobacco

  • Mingqi Zhou
  • Lihua Wu
  • Jing Liang
  • Chen Shen
  • Juan Lin


Inducers of C-repeat binding factor (CBF) expression [inducer of CBF expression 1 (ICE1)-like], being MYC-type transcription factors, play an important role in plant tolerance to low temperatures and other abiotic environmental stresses. These ICEs are the key activators binding to C-repeat/dehydration-responsive elements, triggering the response of CBF signaling and other cold-related genes under cold stress. Our previous report documented a novel ICE transcription factor from Capsella bursa-pastoris, named CbICE53 (GenBank accession no. AY506804), which is responsive to cold, salt stress, and exogenous IAA, GA3, ABA, MeJA, and SA. In the present study, subcellular localization analyses reveal that CbICE53 is specifically localized to the nucleus. Similarly to AtICE1, CbICE53 exhibited slightly increased transcript level under cold induction. Compared with wild-type plants, transformants overexpressing CbICE53 showed increased tolerance to both chilling and freezing temperature according to the index of electrolyte leakage, relative water content and glucose content, and survival rate assay. The expressions of endogenous cold-responsive genes in transgenic tobacco (NtDREB1, NtDREB3, NtERD10a, and NtERD10b) were obviously upregulated only in cold condition. These results suggest that CbICE53 increases plant tolerance to low temperature via activating downstream cold-responsive pathways, which can be considered as a potential candidate for transgenic engineering in plant breeding for cold-tolerant crops.


Capsella bursa-pastoris CbICE53 Cold-responsive genes Cold acclimation Transgenic tobacco 



We acknowledge the financial support from the Natural Science Foundation of China (31170287), the Major Program for the Fundamental Research of Shanghai, China (09JC1401700), the National High Technology Research and Development Program of China (863 Program) (2008AA10Z105), and the National Key Technology R&D Program (2009BADA8B04).

Supplementary material

11032_2012_9744_MOESM1_ESM.docx (298 kb)
Supplementary material 1 (DOCX 299 kb)


  1. Badawi M, Reddy YV, Agharbaoui Z, Tominaga Y, Danyluk J, Sarhan F, Houde M (2008) Structure and functional analysis of wheat ICE (Inducer of CBF Expression) genes. Plant Cell Physiol 49:1237–1249PubMedCrossRefGoogle Scholar
  2. Breton G, Danyluk J, Charron JB, Sarhan F (2003) Expression profiling and bioinformatic analyses of a novel stress-regulated multispanning transmembrane protein family from cereals and Arabidopsis. Plant Physiol 132:64–74PubMedCrossRefGoogle Scholar
  3. Chinnusamy V, Ohta M, Kanrar S, Lee BH, Hong XH, Agarwal M, Zhu JK (2003) ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Gene Dev 17:1043–1054PubMedCrossRefGoogle Scholar
  4. Chinnusamy V, Zhu J, Zhu JK (2007) Cold stress regulation of gene expression in plants. Trends Plant Sci 12:444–451PubMedCrossRefGoogle Scholar
  5. Gilmour SJ, Fowler SG, Thomashow MF (2004) Arabidopsis transcriptional activators CBF1, CBF2, and CBF3 have matching functional activities. Plant Mol Biol 54:767–781PubMedCrossRefGoogle Scholar
  6. Hsieh TH, Lee JT, Yang PT, Chiu LH, Charng YY, Wang YC, Chan MT (2002) Heterology expression of the Arabidopsis C-repeat/dehydration response element binding factor 1 gene confers elevated tolerance to chilling and oxidative stresses in transgenic tomato. Plant Physiol 129:1086–1094PubMedCrossRefGoogle Scholar
  7. Jaglo-Ottosen KR, Gilmour SJ, Zarka DG, Schabenberger O, Thomashow MF (1998) Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance. Science 280:104–106PubMedCrossRefGoogle Scholar
  8. 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 17:287–291PubMedCrossRefGoogle Scholar
  9. 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 45:346–350PubMedCrossRefGoogle Scholar
  10. Miura K, Jin JB, Lee JY, Yoo CY, Stirm V, Miura T, Ashworth EN, Bressan RA, Yun DJ, Hasegawa PM (2007) SIZ1-mediated sumoylation of ICE1 controls CBF3/DREB1A expression and freezing tolerance in Arabidopsis. Plant Cell 19:1403–1414PubMedCrossRefGoogle Scholar
  11. Okawa K, Nakayama K, Kakizaki T, Yamashita T, Inaba T (2008) Identification and characterization of Cor413im proteins as novel components of the chloroplast inner envelope. Plant, Cell Environ 31:1470–1483CrossRefGoogle Scholar
  12. Park JM, Park CJ, Lee SB, Ham B, Shin R, Paek K (2001) Overexpression of the tobacco Tsi1 gene encoding an EREBP/AP2-type transcription factor enhances resistance against pathogen attack and osmotic stress in tobacco. Plant Cell 13:1035–1046PubMedGoogle Scholar
  13. Riechmann JL, Meyerowitz EM (1998) The AP2/EREBP family of plant transcription factor. J Biol Chem 379:633–646Google Scholar
  14. Thomashow MF (1999) Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Annu Rev Plant Physiol Plant Mol Biol 50:571–599PubMedCrossRefGoogle Scholar
  15. Thomashow MF (2001) So what’s new in the field of plant cold acclimation? Lots! Plant Physiol 125:89–93PubMedCrossRefGoogle Scholar
  16. Venketesh S (2008) Properties, potentials, and prospects of antifreeze proteins. Crit Rev Biotechnol 28:57–82PubMedCrossRefGoogle Scholar
  17. Wang XL, Sun XQ, Liu SX, Liu L, Liu XJ, Sun XF, Tang KX (2005) Molecular cloning and characterization of a novel ice gene from Capsella bursa-pastoris. Mol Biol 39:18–25CrossRefGoogle Scholar
  18. Yang W, Liu XD, Chi XJ, Wu CA, Li YZ, Song LL, Liu XM, Wang YF, Wang FW, Zhang C, Liu Y, Zong JM, Li HY (2010) Dwarf apple MbDREB1 enhances plant tolerance to low temperature, drought, and salt stress via both ABA-dependent and ABA-independent pathways. Planta 233:219–229PubMedCrossRefGoogle Scholar
  19. Zhang X, Fowler SG, Cheng HM, Lou YG, Rhee SY, Stockinger EJ, Tomashow MF (2004) Freezing-sensitive tomato has a functional CBF cold response pathway, but a CBF regulon that differs from that of freezing-tolerant Arabidopsis. Plant J 39:905–919PubMedCrossRefGoogle Scholar
  20. Zhou MQ, Wu LH, Shen C, Lin J (2010) Regulation of cold-responsive genes in CBF signaling pathway from Capsella bursa-pastoric induced by ABA, MeJA and SA. J Agric Sci Tech 12:75–80Google Scholar
  21. Zhou MQ, Shen C, Wu LH, Tang KX, Lin J (2011a) CBF-dependent signaling pathway: a key responder to low temperature stress in plants. Crit Rev Biotechnol 31:186–192PubMedCrossRefGoogle Scholar
  22. Zhou MQ, Wu LH, Shen C, Lin J (2011b) A study on the regulation of the expression of cold-responsive genes in CBF signaling pathway from Capsella bursa-pastoric induced by IAA and GA3. J Sichuan Univ (Natural Science Edition) 48:201–205Google Scholar
  23. Zhou MQ, Wu LH, Liang J, Shen C, Lin J (2012) Expression analysis and functional characterization of a novel cold-responsive gene CbCOR15a from Capsella bursa-pastoris. Mol Biol Rep 39:5169–5179PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Mingqi Zhou
    • 1
  • Lihua Wu
    • 1
  • Jing Liang
    • 1
  • Chen Shen
    • 1
  • Juan Lin
    • 1
  1. 1.State Key Laboratory of Genetic Engineering, Institute of Plant Biology, School of Life SciencesFudan UniversityShanghaiChina

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