biologia plantarum

International journal on Plant Life established by Bohumil Nìmec in 1959

Biologia plantarum 55:653, 2011 | DOI: 10.1007/s10535-011-0164-x

Inducible and constitutive expression of HvCBF4 in rice leads to differential gene expression and drought tolerance

T. Lourenço1, N. Saibo1, R. Batista1,2, C. Pinto Ricardo1, M. M. Oliveira1,*
1 ITQB-Universidade Nova de Lisbo, Laboratório de Genómica de Plantas em Stress, Oeiras, Portugal
2 Instituto Nacional de Saúde Dr. Ricardo Jorge, Lisboa, Portugal

The effects of the ectopic expression of a barley transcription factor (HvCBF4) under the control of a constitutive (maize Ubi1) or a stress-inducible (Arabidopsis RD29A) promoter in the abiotic stress response in rice (Oryza sativa L.) was investigated. The transformed plants were analyzed both at molecular and physiological level and the AtRD29A::HvCBF4 plants were further analyzed using the GeneChip® rice genome array under control conditions. Only the plants constitutively expressing HvCBF4 have shown increased survival to drought stress, but not to cold or high-salinity. These plants have also shown better photosynthetic capacity, as determined by chlorophyll fluorescence. Plants expressing AtRD29A::HvCBF4 did not show increased survival to any of the stresses applied. However in the GeneChip® microarray, these plants have shown up-regulation of many stress-responsive genes (> 400) as compared to non-transformed plants. Interestingly, RT-PCR analysis revealed not only differential gene expression between roots and shoots, but also between transgenic lines with the different promoters. Our results indicate that different HvCBF4 expression levels resulted in different transcriptomes and drought tolerance. Given that AtRD29A::HvCBF4 plants did not show increased tolerance to any of the imposed stresses, we may conclude that this promoter may be inappropriate for rice transformation aiming for enhanced abiotic stress tolerance.

Keywords: abiotic stress; chlorophyll fluorescence; microarray analysis; Oryza sativa; transcription factors; transformation

Received: April 29, 2010; Accepted: August 16, 2010; Published: December 1, 2011  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Lourenço, T., Saibo, N., Batista, R., Pinto Ricardo, C., & Oliveira, M.M. (2011). Inducible and constitutive expression of HvCBF4 in rice leads to differential gene expression and drought tolerance. Biologia plantarum, 55(4), 653. doi: 10.1007/s10535-011-0164-x
Share...
Download citation
  • BibTeX (.bib)
  • Bookends (.ris)
  • EasyBib (.ris)
  • EndNote (.enw)
  • EndNote 8 (.xml)
  • ISI WoS (.isi)
  • Medlars (.medlars)
  • Mendeley (.ris)
  • MODS (.xml)
  • Papers (.ris)
  • RefWorks (.txt)
  • RefManager (.ris)
  • RIS (.ris)
  • MS Word (.xml)
  • Zotero (.ris)
    • Open full article

    References

    1. Agarwal, P.K., Jha, B.: Transcription factors in plants and ABA dependent and independent abiotic stress signalling. - Biol. Plant. 54: 201-212, 2010. Go to original source...
    2. Baker, S.S., Wilhelm, K.S., Thomashow, M.F.: 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, 1994. Go to original source...
    3. Barr, H.D., Weatherley, P.E.: A re-examination of the relative turgidity technique for estimating water deficit in leaves. - Aust. J. biol. Sci. 15: 413-428, 1962. Go to original source...
    4. Behnam, B., Kikuchi, A., Celebi-Toprak, F., Kasuga, M., Yamaguchi-Shinozaki, K., Watanabe, K.N.: Arabidopsis rd29A::DREB1A enhances freezing tolerance in transgenic potato. - Plant Cell Rep. 26: 1275-1282, 2007. Go to original source...
    5. Bhatnagar-Mathur, P., Devi, M.J., Reddy, D.S., Lavanya, M., Vadez, V., Serraj, R., Yamaguchi-Shinozaki, K., Sharma, K.K.: Stress-inducible expression of At DREB1A in transgenic peanut (Arachis hypogaea L.) increases transpiration efficiency under water-limiting conditions. - Plant Cell Rep. 26: 2071-2082, 2007. Go to original source...
    6. Boyer, J.S.: Plant productivity and environment. - Science 218: 443-448, 1982. Go to original source...
    7. Choi, D.W., Rodriguez, E.M., Close, T.J.: Barley Cbf3 gene identification, expression pattern, and map location. - Plant Physiol. 129: 1781-1787, 2002. Go to original source...
    8. Christensen, A.H., Quail, P.H.: Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants. - Transgenic Res. 5: 213-218, 1996. Go to original source...
    9. Cullings, K.W.: Design and testing of a plant-specific PCR primer for ecological and evolutionary studies. - Mol. Ecol. 1: 233-240, 1992. Go to original source...
    10. Doyle, J.J., Doyle, J.L.: A rapid DNA isolation procedure for small quantities of fresh leaf tissue. - Phytochem. Bull. 19: 11-15, 1987.
    11. Dubouzet, J.G., Sakuma, Y., Ito, Y., Kasuga, M., Dubouzet, E.G., Miura, S., Seki, M., Shinozaki, K., Yamaguchi-Shinozaki, K.: 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, 2003. Go to original source...
    12. Fryer, M.J., Andrews, J.R., Oxborough, K., Blowers, D.A., Baker, N.R.: Relationship between CO2 assimilation, photosynthetic electron transport, and active O2 metabolism in leaves of maize in the field during periods of low temperature. - Plant Physiol. 116: 571-580, 1998. Go to original source...
    13. Garg, A.K., Kim, J.K., Owens, T.G., Ranwala, A.P., Choi, Y.D., Kochian, L.V., Wu, R.J.: Trehalose accumulation in rice plants confers high tolerance levels to different abiotic stresses. - Proc nat. Acad. Sci. USA 99: 15898-15903, 2002. Go to original source...
    14. Gilmour, S.J., Sebolt, A.M., Salazar, M.P., Everard, J.D., Thomashow, M.F.: Overexpression of the Arabidopsis CBF3 transcriptional activator mimics multiple biochemical changes associated with cold acclimation. - Plant Physiol. 124: 1854-1865, 2000. Go to original source...
    15. Gilmour, S.J., Zarka, D.G., Stockinger, E.J., Salazar, M.P., Houghton, J.M., Thomashow, M.F.: 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, 1998. Go to original source...
    16. Haake, V., Cook, D., Riechmann, J.L., Pineda, O., Thomashow, M.F., Zhang, J.Z.: Transcription factor CBF4 is a regulator of drought adaptation in Arabidopsis. - Plant Physiol. 130: 639-648, 2002. Go to original source...
    17. Hiei, Y., Ohta, S., Komari, T., Kumashiro, T.: Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. - Plant J. 6: 271-282, 1994. Go to original source...
    18. Hsieh, T.H., Lee, J.T., Charng, Y.Y., Chan, M.T.: Tomato plants ectopically expressing Arabidopsis CBF1 show enhanced resistance to water deficit stress. - Plant Physiol. 130: 618-626, 2002. Go to original source...
    19. Hu, X.J., Zhang, Z.B., Xu, P., Fu, Z.Y., Hu, S.B., Song, W.Y.: Multifunctional genes: the cross-talk among the regulation networks of abiotic stress responses. - Biol. Plant. 54: 213-223, 2010. Go to original source...
    20. Ito, Y., Katsura, K., Maruyama, K., Taji, T., Kobayashi, M., Seki, M., Shinozaki, K., Yamaguchi-Shinozaki, K.: Functional analysis of rice DREB1/CBF-type transcription factors involved in cold-responsive gene expression in transgenic rice. - Plant Cell Physiol. 47: 141-153, 2006. Go to original source...
    21. Jaglo, K.R., Kleff, S., Amundsen, K.L., Zhang, X., Haake, V., Zhang, J.Z., Deits, T., Thomashow, M.F.: 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, 2001. Go to original source...
    22. Jaglo-Ottosen, K.R., Gilmour, S.J., Zarka, D.G., Schabenberger, O., Thomashow, M.F.: Arabidopsis CBF1 overexpression induces COR genes and enhances freezing tolerance. - Science 280: 104-106, 1998. Go to original source...
    23. Kasuga, M., Liu, Q., Miura, S., Yamaguchi-Shinozaki, K., Shinozaki, K.: Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor. - Nat. Biotechnol. 17: 287-291, 1999. Go to original source...
    24. Kasuga, M., Miura, S., Shinozaki, K., Yamaguchi-Shinozaki, K.: 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, 2004. Go to original source...
    25. Liu, Q., Kasuga, M., Sakuma, Y., Abe, H., Miura, S., Yamaguchi-Shinozaki, K., Shinozaki, K.: 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, 1998. Go to original source...
    26. Maxwell, K., Johnson, G.N.: Chlorophyll fluorescence - a practical guide. - J. exp. Bot. 51: 659-668, 2000. Go to original source...
    27. Nakashima, K., Yamaguchi-Shinozaki, K.: Regulons involved in osmotic stress-responsive and cold stress-responsive gene expression in plants. - Physiol. Plant. 126: 62-71, 2006. Go to original source...
    28. Oh, S.J., Kwon, C.W., Choi, D.W., Song, S.I., Kim, J.K.: Expression of barley HvCBF4 enhances tolerance to abiotic stress in transgenic rice. - Plant Biotechnol. J. 5: 646-656, 2007. Go to original source...
    29. Oh, S.J., Song, S.I., Kim, Y.S., Jang, H.J., Kim, S.Y., Kim, M., Kim, Y.K., Nahm, B.H., Kim, J.K.: Arabidopsis CBF3/DREB1A and ABF3 in transgenic rice increased tolerance to abiotic stress without stunting growth. - Plant Physiol. 138: 341-351, 2005. Go to original source...
    30. Ono, A., Izawa, T., Chua, N.H., Shimamoto, K.: The rab16B promoter of rice contains two distinct abscisic acidresponsive elements. - Plant Physiol. 112: 483-491, 1996. Go to original source...
    31. Pellegrineschi, A., Reynolds, M., Pacheco, M., Brito, R.M., Almeraya, R., Yamaguchi-Shinozaki, K., Hoisington, D.: Stress-induced expression in wheat of the Arabidopsis thaliana DREB1A gene delays water stress symptoms under greenhouse conditions. - Genome 47: 493-500, 2004. Go to original source...
    32. Pino, M.T., Skinner, J.S., Park, E.J., Jeknic, Z., Hayes, P.M., Thomashow, M.F., Chen, T.H.: Use of a stress inducible promoter to drive ectopic AtCBF expression improves potato freezing tolerance while minimizing negative effects on tuber yield. - Plant Biotechnol. J. 5: 591-604, 2007. Go to original source...
    33. Rueb, S., Leneman, M., Schilperoort, R.A., Hensgens, L.A.M.: Efficient plant regeneration through somatic embryogenesis from callus induced on mature rice embryos (Oryza sativa L.). - Plant Cell Tissue Organ Cult. 36: 259-264, 1994. Go to original source...
    34. Sakamoto, H., Maruyama, K., Sakuma, Y., Meshi, T., Iwabuchi, M., Shinozaki, K., Yamaguchi-Shinozaki, K.: Arabidopsis Cys2/His2-type zinc-finger proteins function as transcription repressors under drought, cold, and highsalinity stress conditions. - Plant Physiol. 136: 2734-2746, 2004. Go to original source...
    35. Savitch, L.V., Allard, G., Seki, M., Robert, L.S., Tinker, N.A., Huner, N.P., Shinozaki, K., Singh, J.: The effect of overexpression of two Brassica CBF/DREB1-like transcription factors on photosynthetic capacity and freezing tolerance in Brassica napus. - Plant Cell Physiol. 46: 1525-1539, 2005. Go to original source...
    36. Seki, M., Narusaka, M., Abe, H., Kasuga, M., Yamaguchi-Shinozaki, K., Carninci, P., Hayashizaki, Y., Shinozaki, K.: Monitoring the expression pattern of 1300 Arabidopsis genes under drought and cold stresses by using a full-length cDNA microarray. - Plant Cell 13: 61-72, 2001. Go to original source...
    37. Skinner, J.S., Von Zitzewitz, J., Szucs, P., Marquez-Cedillo, L., Filichkin, T., Amundsen, K., Stockinger, E.J., Thomashow, M.F., Chen, T.H., Hayes, P.M.: Structural, functional, and phylogenetic characterization of a large CBF gene family in barley. - Plant mol. Biol. 59: 533-551, 2005. Go to original source...
    38. Stockinger, E.J., Gilmour, S.J., Thomashow, M.F.: 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. nat. Acad. Sci. USA 94: 1035-1040, 1997. Go to original source...
    39. Tamura, K., Dudley, J., Nei, M., Kumar, S.: MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. - Mol. Biol. Evol. 24: 1596-1599, 2007. Go to original source...
    40. Thomashow, M.F.: plant cold acclimation: freezing tolerance genes and regulatory mechanisms. - Annu. Rev. Plant Physiol. Plant mol. Biol. 50: 571-599, 1999. Go to original source...
    41. Umezawa, T., Fujita, M., Fujita, Y., Yamaguchi-Shinozaki, K., Shinozaki, K.: Engineering drought tolerance in plants: discovering and tailoring genes to unlock the future. - Curr. Opin. Biotechnol. 17: 113-122, 2006. Go to original source...
    42. Vogel, J.T., Zarka, D.G., Van Buskirk, H.A., Fowler, S.G., Thomashow, M.F.: Roles of the CBF2 and ZAT12 transcription factors in configuring the low temperature transcriptome of Arabidopsis. - Plant J. 41: 195-211, 2005. Go to original source...
    43. Wang, W., Vinocur, B., Altman, A.: Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. - Planta 218: 1-14, 2003. Go to original source...
    44. Xue, G.P.: The DNA-binding activity of an AP2 transcriptional activator HvCBF2 involved in regulation of lowtemperature responsive genes in barley is modulated by temperature. - Plant J. 33: 373-383, 2003. Go to original source...
    45. Yamaguchi-Shinozaki, K., Shinozaki, K.: 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, 1994. Go to original source...
    46. Yoshida, S., Forno, D.A., Cock, J.H., Gomez, K.A.: Laboratory Manual for Physiological Studies of Rice. - International Rice Research Institute, Manila 1976.
    47. Zhang, J.Z., Creelman, R.A., Zhu, J.K.: From laboratory to field. Using information from Arabidopsis to engineer salt, cold, and drought tolerance in crops. - Plant Physiol. 135: 615-621, 2004. Go to original source...

    Return to the content