Biologia plantarum 56:484-492, 2012 | DOI: 10.1007/s10535-012-0059-5

Isolation and expression of a cold-responsive gene PtCBF in Poncirus trifoliata and isolation of citrus CBF promoters

L. G. He1, H. L. Wang1, D. C. Liu1, Y. J. Zhao1, M. Xu2, M. Zhu1, G. Q. Wei1, Z. H. Sun1,2,*
1 College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan, P.R. China
2 Institute of Fruit and Tea, Hubei Academy of Agricultural Sciences, Wuhan, P.R. China

C-repeat/dehydration-responsive element binding factor (CBF) plays important roles in cold response network in plants. Here, one member of CBF coding gene family in trifoliate orange (Poncirus trifoliata), designated as PtCBF, was isolated. Semi-quantitative reverse transcription-polymerase chain reactions showed up-regulation of PtCBF not only under low temperature but also induced by abscisic acid. Additionally, the CBF genomic fragments in four citrus species including trifoliate orange, sweet orange (Citrus sinensis), pummel (Citrus grandis) and rough lemon (Citrus jambhiri) were isolated with complete open reading frames. According to the results of alignment analysis between full length cDNA and genomic DNA sequences in trifoliate orange, there were no introns in PtCBF. Moreover, the results of multiple sequence alignment analysis and phylogenetic analysis on putative protein sequences suggested that the AP2 DNA binding domains and CBF signature sequences were highly conserved in four citrus CBF proteins. Finally, the CBF promoters in above citrus species were isolated, which provides some information concerning promoter function.

Keywords: abscisic acid; Citrus grandis; Citrus jambhiri; Citrus sinensis; low temperature; trifoliate orange
Subjects: abscisic acid; cold-responsive genes; PCR; CBF/DREB; gene construct; temperature - low; cis-acting elements; phylogenetic tree; promoters
Species: Citrus grandis; Citrus jambhiri; Citrus sinensis; Poncirus trifoliata

Received: January 20, 2010; Accepted: March 30, 2010; Published: September 1, 2012Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
He, L.G., Wang, H.L., Liu, D.C., Zhao, Y.J., Xu, M., Zhu, M., Wei, G.Q., & Sun, Z.H. (2012). Isolation and expression of a cold-responsive gene PtCBF in Poncirus trifoliata and isolation of citrus CBF promoters. Biologia plantarum, 56(3), 484-492. doi: 10.1007/s10535-012-0059-5.
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. Badawi, M., Danyluk, J., Boucho, B., Houde, M., Sarhan, F.: The CBF gene family in hexaploid wheat and its relationship to the phylogenetic complexity of cereal CBFs. - Mol. Genet. Genomics 277: 533-554, 2007. 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. Benedict, C., Skinner, J.S., Meng, R., Chang, Y., Bhalerao, R., Huner, N.P., Finn, C.E., Chen, T.H., Hurry, V.: The CBF1-dependent low temperature signalling pathway, regulon, and increase in freeze tolerance are conserved in Populus spp.. - Plant Cell Environ. 29: 1259-1272, 2006. Go to original source...
    4. Cai, Q.Y., Moore, G.A., Guy, C.L.: An unusual Group 2 LEA gene family in citrus responsive to low temperature. - Plant mol. Biol. 29: 11-23, 1995. Go to original source...
    5. 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. Go to original source...
    6. Cheng, Y.J., Guo, W.W., Yi, H.L., Pang, X.M., Deng, X.X.: An efficient protocol for genomic DNA extraction from Citrus species. - Plant mol. Biol. Rep. 21(Suppl.): 177a-177g, 2003. Go to original source...
    7. El Kayal, W., Navarro, M., Marque, G., Keller, G., Marque, C., Teulieres, C.: Expression profile of CBF-like transcriptional factor genes from Eucalyptus in response to cold. - J. exp. Bot. 57: 2455-2469, 2006. Go to original source...
    8. Fowler, S.G., Cook, D., Thomashow, M.F.: Low temperature induction of Arabidopsis CBF1, 2 and 3 is gated by the circadian clock. - Plant Physiol. 137: 961-968, 2005. Go to original source...
    9. Gao, M.J., Allard, G., Byass, L., Flanagan, A.M., Singh, J.: Regulation and characterization of four CBF transcription factors from Brassica napus. - Plant Mol. Biol. 49: 459-471, 2002. Go to original source...
    10. Gilliland, L.U., Pawloski, L.C., Kandasamy, K., Meaghe, R.B.: Arabidopsis actin gene ACT7 plays an essential role in germination and root growth. - Plant J. 33: 319-328, 2003. Go to original source...
    11. Gilmour, S.J., Fowler, S.G., Thomashow, M.F.: Arabidopsis transcriptional activators CBF1, CBF2, and CBF3 have matching functional activities. - Plant mol. Biol. 54: 767-781, 2004. Go to original source...
    12. 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.
    13. Gutha, L.R., Reddy, A.R.: Rice DREB1B promoter shows distinct stress-specific responses, and the overexpression of cDNA in tobacco confers improved abiotic and biotic stress tolerance. - Plant mol. Biol. 68: 533-555, 2008. Go to original source...
    14. Hellens, R.P., Allan, A.C., Friel, E.N., Bolitho, K., Grafton, K., Templeton, M.D., Karunairetnam, S., Gleave, A.P., Laing, W.A.: Transient expression vectors for functional genomics, quantification of promoter activity and RNA silencing in plants. - Plant Methods 1: 13, 2005. Go to original source...
    15. 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...
    16. 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. - Natur. Biotechnol. 17: 287-291, 1999. Go to original source...
    17. Kim, H.J., Kim, Y.K., Park, J.Y., Kim, J.: Light signaling mediated by phytochrome plays an important role in coldinduced gene expression through the C-repeat/dehydration responsive element (C/DRE) in Arabidopsis thaliana. - Plant J. 29: 693-704, 2002. Go to original source...
    18. Knight, H., Zarka, D.G., Okamoto, H., Thomashow, M.F., Knight, M.R.: Abscisic acid induces CBF gene transcription and subsequent induction of cold-regulated genes via the CRT promoter element. - Plant Physiol. 135: 1710-1717, 2004. Go to original source...
    19. Liu, Q., Xu, J., Liu, Y.Z., Zhao, X.L., Deng, X.X., Guo, L.L., Gu, J.Q.: A novel bud mutation that confers abnormal patterns of lycopene accumulation in sweet orange fruit (Citrus sinensis L. Osbeck). - J. exp. Bot. 58: 4161-4171, 2007. Go to original source...
    20. Meng, S.S., Dane, F., Si, Y., Ebel, R., Zhang, C.K.: Gene expression analysis of cold treated versus cold acclimated Poncirus trifoliata. - Euphytica 164: 209-219, 2008. Go to original source...
    21. Novillo, F., Medina, J., Salinas, J.: Arabidopsis CBF1 and CBF3 have a different function than CBF2 in cold acclimation and define different gene classes in the CBF regulon. - Proc. nat. Acad. Sci. USA 104: 21002-21007, 2007. Go to original source...
    22. Qin, F., Sakuma, Y., Li, J., Liu, Q., Li, Y.Q., Shinozaki, K., Yamaguchi-Shinozaki, K.: Cloning and functional analysis of a novel DREB1/CBF transcription factor involved in cold-responsive gene expression in Zea mays L.. - Plant Cell Physiol. 45: 1042-1052, 2004. Go to original source...
    23. Seo, E., Lee, H., Jeon, J., Park, H., Kim, J., Noh, Y.S., Lee, I.: Crosstalk between cold response and flowering in ARABIDOPSIS is mediated through the flowering-time gene SOC1 and its upstream negative regulator FLC. - Plant Cell 21: 3185-3197, 2009. Go to original source...
    24. Svensson, J.T., Crosatti, C., Campoli, C., Bassi, R., Stanca, A.M., Close, T.J., Cattivelli, L.: Transcriptome analysis of cold acclimation in barley albina and xantha mutants. - Plant Physiol. 141: 257-270, 2006. Go to original source...
    25. Talon, M., Gmitter, F.G.: Citrus genomics. - Int. J. Plant Genomics 2008: 1-17, 2008. Go to original source...
    26. Terauchi, R., Kahl, G.: Rapid isolation of promoter sequences by TAIL-PCR: the 5'-flanking regions of Pal and Pgi genes from yams (Dioscorea). - Mol. gen. Genet. 263: 554-560, 2000. Go to original source...
    27. Thomashow, M.F., Gilmour, S.J., Stockinger, E.J., Jaglo-Ottosen, K.R., Zarka, D.G.: Role of the Arabidopsis CBF transcriptional activators in cold acclimation. - Physiol. Plant. 112: 171-175, 2001. Go to original source...
    28. Wang, H.L., Tao, J.J., He, L.G., Zhao, Y.J., Xu, M, Liu, D.C., Sun, Z.H.: cDNA cloning and expression analysis of a Poncirus trifoliata CBF gene. - Biol. Plant. 53: 625-630, 2009. Go to original source...
    29. Wang, Q.Y., Guan, Y.C., Wu, Y.R., Chen, H.L., Chen, F., Chu, C.C. Overexpression of a rice OsDREB1F gene increases salt, drought, and low temperature tolerance in both Arabidopsis and rice. - Plant mol. Biol. 67: 589-602, 2008. Go to original source...
    30. Xiao, H.G., Siddiqua, M., Braybrook, S., Nassuth, A.: Three grape CBF/DREB1 genes respond to low temperature, drought and abscisic acid. - Plant Cell Environ. 29: 1410-1421, 2006. Go to original source...
    31. Xue, G.P.: Characterisation of the DNA-binding profile of barley HvCBF1 using an enzymatic method for rapid, quantitative and high-throughput analysis of the DNAbinding activity. - Nucl. Acids Res. 30(Suppl.): e77, 2002. Go to original source...
    32. Zhang, C.K., Lang, P., Dane, F., Ebel, R.C., Singh, N.K., Locy, R.D., Dozier, W.A.: Cold acclimation induced genes of trifoliate orange (Poncirus trifoliata). - Plant Cell Rep. 23: 764-769, 2005. Go to original source...
    33. Zhang, X., Fowler, S.G., Cheng, H.M., Lou, Y.G., Rhee, S.Y., Stockinger, E.J., Thomashow, M.F.: 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: 905-919, 2004. Go to original source...

    Return to the content