Advertisement

Biologia Plantarum

, 53:625 | Cite as

cDNA cloning and expression analysis of a Poncirus trifoliata CBF gene

  • H. L. Wang
  • J. J. Tao
  • L. G. He
  • Y. J. Zhao
  • M. Xu
  • D. C. Liu
  • Z. H. Sun
Original Papers

Abstract

The transcription factors CBF/DREB play an important role during low temperature, drought and high-salt stress in higher plants. A new CBF (CRT/DRE binding factor) gene was cloned from trifoliate orange [Poncirus trifoliata (L.) Raf.] by RT-PCR with degenerate primers and rapid amplification of cDNA ends (RACE) techniques. The full-length cDNA of CBF gene from trifoliate orange (designated as Ptcbfb) was 847 bp containing a 732 bp open reading frame (ORF), encoding a 243 amino acid protein. The predicted protein (designated as PtCBFb) had over 60 % identity to CBFs from some other plant species. Bioinformatical analysis showed that PtCBFb contained N-terminal bipartite nuclear targeting sequence, potential C-terminal acid domain and high conserved AP2 domain. Some other loci such as phosphorylation sites of several protein kinases, N-myristoylation site, tyrosine sulfation site and amidation site were also conserved in PtCBFb. Predicted three-dimentional structure of PtCBFb was similar to CBF from Arabidopsis thaliana. Expression pattern analysis revealed Ptcbfb expression in every tested organ, and Ptcbfb was cold induced.

Additional key words

full-length cDNA PtCBFb RACE trifoliate orange 

Abbreviations

CBF/DREB

CRT/DRE binding factor

ORF

open reading frame

RACE

rapid amplification of cDNA ends

TMHs

transmembrane helix

NLS

nuclear localization signal

RT

reverse transcript

References

  1. Allen, M.D., Yamasaki, K., Ohme-Takagi, M.: A novel mode of DNA recognition by a beta-sheet revealed by the solution structure of the GCC-box binding domain in complex with DNA. — EMBO J. 17: 5484–5496, 1998.CrossRefPubMedGoogle Scholar
  2. Champ, K.I., Febres, V.J., Moore, G.A.: The role of CBF transcriptional activators in two Citrus species (Poncirus and Citrus) with contrasting levels of freezing tolerance. — Physiol. Plant. 129: 529–541, 2007.CrossRefGoogle Scholar
  3. Choi, D.W., Rodriguez, E.M., Close, T.J.: Barley Cbf3 gene identification, expression pattern, and map location. — Plant Physiol. 129: 1781–1787, 2002.CrossRefPubMedGoogle Scholar
  4. Francia, E., Rizza, F., Cattivelli, L., Stanca, A.M., Galiba, G., Toth, B., Hayes, P.M., Skinner, J.S., Pecchioni, N.: Two loci on chromosome 5H determine low-temperature tolerance in a ‘Nure’ (winter) × ‘Tremois’ (spring) barley map. — Theor. appl. Genet. 108: 670–680, 2004.CrossRefPubMedGoogle Scholar
  5. Gamboa, M.C., Rasmussen-Poblete, S.: Isolation and characterization of a cDNA encoding a CBF transcription factor from E. globulus. — Plant Physiol. Biochem. 45: 1–5, 2007.CrossRefPubMedGoogle Scholar
  6. 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.CrossRefPubMedGoogle Scholar
  7. Liu, J.J., Goh, C.J., Loh, C.S., Liu, P., Pua, E.C.: A method for isolation of total RNA from fruit tissues of banana. — Plant mol. Biol. Rep. 16: 1–6, 1998a.CrossRefGoogle Scholar
  8. 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, 1998b.CrossRefPubMedGoogle Scholar
  9. Shinwari, Z.K., Nakashima, K., Miura, S., Kasuga, M., Seki, M., Yamaguchi-Shinozaki, K., Shinozaki, K.: An Arabidopsis gene family encoding DRE/CRT binding proteins involved in low-temperature-responsive gene expression. — Biochem. biophys. Res. Commun. 250: 161–170, 1998.CrossRefPubMedGoogle Scholar
  10. Thomashow, M.F.: Molecular genetics of cold acclimation in higher plants. — Adv Genet. 28: 99–131, 1990.CrossRefGoogle Scholar
  11. Thomashow, M.F.: So What’s new in the field of plant cold acclimation? Lots! — Plant Physiol. 125: 89–93, 2001.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • H. L. Wang
    • 1
  • J. J. Tao
    • 1
  • L. G. He
    • 1
  • Y. J. Zhao
    • 1
  • M. Xu
    • 1
  • D. C. Liu
    • 1
  • Z. H. Sun
    • 1
  1. 1.Key Laboratory of Horticultural Plant Biology, Ministry of Education, Citrus Research InstituteHuazhong Agricultural UniversityWuhanP.R. China

Personalised recommendations