Skip to main content
Log in

Overexpression of a ItICE1 gene from Isatis tinctoria enhances cold tolerance in rice

  • Published:
Molecular Breeding Aims and scope Submit manuscript

Abstract

ItICE1, a ICE1-like gene, was isolated from a cDNA library from cold-treated woad (Isatis tinctoria L.) tissues. Expression analysis revealed that the ItICE1 gene was expressed constitutively and was predominant in the leaves of woad seedlings and that its mRNA accumulation was altered by salt stress and abscisic acid application, but not by dehydration and cold stresses. The transgenic rice lines overexpressing ItICE1 showed no growth retardation under normal growth conditions as well as enhanced tolerance to cold stress. Physiological assays showed that ItICE1 not only increased the accumulation of free proline and chlorophyll in transgenic rice lines under cold stress, but also reduced malondialdehyde content and electrolyte leakage. The analysis of gene expression in transgenic rice lines indicated that the maize ubiquitin promoter could respond to cold stress and upregulate ItICE1 gene expression level under its control. Under cold stress conditions, transgenic lines had a remarkably increased expression of OsDREB1A, J013078A14, 001-125-G03, 001-023-B08 and J023042N13 compared to wild-type plants (P < 0.05), implying that ItICE1 functions in the CBF/DREB1 cold-response pathway. These results demonstrate that ItICE1 plays an important regulatory role in the improvement of tolerance to cold stress in rice and is potentially useful for improving the cold tolerance of other plants.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • 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(8):1237–1249

    Article  PubMed  CAS  Google Scholar 

  • Baker SS, Wilhem KS, Thomashow MF (1994) The 50-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  PubMed  CAS  Google Scholar 

  • Chen JQ, Meng XP, Zhang Y, Xia M, Wang XP (2008) Overexpression of OsDREB genes lead to enhanced drought tolerance in rice. Biotechnol Lett 30:2191–2198

    Article  PubMed  Google Scholar 

  • Chinnusamy V, Ohta M, Kanrar S, Lee BH, Hong X, Agarwal M, Zhu JK (2003) ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Genes Dev 17:1043–1054

    Article  PubMed  CAS  Google Scholar 

  • Chinnusamy V, Zhu JK, Sunkar R (2010) Gene regulation during cold stress acclimation in plants. Methods Mol Biol 639:39–55

    Article  PubMed  CAS  Google Scholar 

  • Dong CH, Agarwal M, Zhang Y, Xie Q, Zhu JK (2006) The negative regulator of plant cold responses, HOS1, is a RING E3 ligase that mediates the ubiquitination and degradation of ICE1. Proc Natl Acad Sci USA 103:8281–8286

    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 

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

    Article  CAS  Google Scholar 

  • Hiei Y, Komari T, Kubo T (1997) Transformation of rice mediated by Agrobacterium tumefaciens. Plant Mol Biol 35:205–218

    Article  PubMed  CAS  Google Scholar 

  • Hodges DM, DeLong JM, Forney CF, Prange RK (1999) Improving the thiobarbituric acid-reactive-substances assay for estimating lipid peroxidation in plant tissues containing anthocyanin and other interfering compounds. Planta 207:604–611

    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  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  PubMed  CAS  Google Scholar 

  • Jiang FL, Wang F, Wu Z, Li Y, Shi GJ, Hu JD, Hou XL (2011) Components of the Arabidopsis CBF cold responsive pathway are conserved in non-heading Chinese Cabbage. Plant Mol Biol Rep 29:525–532

    Article  CAS  Google Scholar 

  • Karam BS, Rhonda CF, Luis OS (2002) Transcription factors in plant defense and stress responses. Curr Opin Plant Biol 5:430–436

    Article  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  PubMed  CAS  Google Scholar 

  • Knight H, Veale EL, Warren GJ, Knight MR (1999) The sfr6 mutation in Arabidopsis suppresses low-temperature induction of genes dependent on the CRT/DRE sequence motif. Plant Cell 11:875–886

    PubMed  CAS  Google Scholar 

  • Li F, Guo SY, Zhao YA, Chen DZ, Chong K, Xu YY (2010) Overexpression of a homopeptide repeat-containing bHLH protein gene (OrbHLH001) from Dongxiang Wild Rice confers freezing and salt tolerance in transgenic Arabidopsis. Plant Cell Rep 29:977–986

    Article  PubMed  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

    PubMed  CAS  Google Scholar 

  • Medina J, Bargues M, Terol J, Perez-Alonso M, Salinas J (1999) The Arabidopsis CBF gene family is composed of three genes encoding AP2 domain-containing proteins whose expression Is regulated by low temperature but not by abscisic acid or dehydration. Plant Physiol 119:463–470

    Article  PubMed  CAS  Google Scholar 

  • Minty A, Dumont X, Kaghad M, Caput D (2000) Covalent modification of p73 by SUMO-1. Two-hybrid screening with p73 identifies novel SUMO-1-interacting proteins and a SUMO-1 interaction motif. J Biol Chem 275:36316–36323

    Article  PubMed  CAS  Google Scholar 

  • Miura K, Jin JB, Lee J, 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 19:1403–1414

    Article  PubMed  CAS  Google Scholar 

  • Morsy MR, Almutairi AM, Gibbons J, Yun SJ, de Los Reyes BG (2005) The OsLti6 genes encoding low-molecular weight membrane proteins are differentially expressed in rice cultivars with contrasting sensitivity to low temperature. Gene 344:171–180

    Article  PubMed  CAS  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 

  • Nanjo T, Kobayashi M, Yoshiba Y, Kakubari Y, Yamaguchi-Shinozaki K, Shinozaki K (1999) Antisense suppression of proline degradation improves tolerance to freezing and salinity in Arabidopsis thaliana. FEBS Lett 461:205–210

    Article  PubMed  CAS  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  PubMed  CAS  Google Scholar 

  • Park HC, Kim H, Koo SC, Park HJ, Cheong MS, Hong H, Baek D, Chung WS, Kim DH, Bressan RA, Lee SY, Bohnert HJ, Yun DJ (2010) Functional characterization of the SIZ/PIAS-type SUMO E3 ligases, OsSIZ1 and OsSIZ2 in rice. Plant, Cell Environ 33:1923–1934

    Article  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Perales L, Penarrubiab L, Cornejoa MJ (2008) Induction of a polyubiquitin genepromoter by dehydration stresses in transformed rice cells. J Plant Physiol 165(2):159–171

    Article  PubMed  CAS  Google Scholar 

  • Santos E, Remy S, Thiry E, Windelinckx S, Swennen R, Sagi L (2009) Characterization and isolation of a T-DNA tagged banana promoter active during in vitro culture and low temperature stress. BMC Plant Biol 9:77

    Article  PubMed  Google Scholar 

  • Sharma P, Sharma N, Deswal R (2005) The molecular biology of the low-temperature response in plants. BioEssays 27:1048–1059

    Article  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 Biophy Res Commun 250:161–170

    Article  CAS  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 

  • Takahashi T, Gasch A, Nishizawa N, Chua NH (1995) The DIMINUTO gene of Arabidopsis is involved in regulating cell elongation. Genes Dev 9:97–107

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Tian H, Li XP, Zhou HL, Zhang JS, Gong ZZ (2005) Chen SY (2005) OsDREB4 genes rice encode AP2-containing proteins that bind specifically to the dehydration responsive element. J Integr Plant Biol 47(4):467–476

    Article  CAS  Google Scholar 

  • Tian Y, Zhang HW, Pan XW, Chen XL, Zhang ZJ, Lu XY, Huang RF (2011) Overexpression of ethylene response factor TERF2 confers cold tolerance in rice seedlings. Transgenic Res 20:857–866

    Article  PubMed  CAS  Google Scholar 

  • Toledo-Ortiz G, Huq E, Quail PH (2003) The Arabidopsis basic/helix-loop-helix transcription factor family. Plant Cell 15:1749–1770

    Article  PubMed  CAS  Google Scholar 

  • Wang QY, Guan YC, Wu YR, Chen HL, Chen F, Chu CC (2008) Overexpression of a rice OsDREB1F gene increases salt, drought, and low temperature tolerance in both Arabidopsis and rice. Plant Mol Biol 67:589–602

    Article  PubMed  CAS  Google Scholar 

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

    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 

Download references

Acknowledgments

This study was supported by the National Natural Science Foundation of China (No. 30971845) and the National Science and Technology Support Program for the 12th five year plan of China (No. 2011BAD35B02).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Zhengjin Xu.

Additional information

Dianjun Xiang, Lili Man, and Kuide Yin contributed equally to this article.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 2031 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Xiang, D., Man, L., Yin, K. et al. Overexpression of a ItICE1 gene from Isatis tinctoria enhances cold tolerance in rice. Mol Breeding 32, 617–628 (2013). https://doi.org/10.1007/s11032-013-9894-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11032-013-9894-0

Keywords

Navigation