Skip to main content
Log in

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

  • Published:
Biologia Plantarum

Abstract

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.

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.

Similar content being viewed by others

Abbreviations

ABA:

abscisic acid

CBF:

C-repeat/dehydration-responsive element binding factor

RT-PCR:

semi-quantitative reverse transcription-polymerase chain reaction

References

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  Google Scholar 

  • 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.

    Article  PubMed  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • Talon, M., Gmitter, F.G.: Citrus genomics. — Int. J. Plant Genomics 2008: 1–17, 2008.

    Article  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

  • 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.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Z. H. Sun.

Additional information

Acknowledgements: This research was supported by the National Natural Science Foundation of China (Nos. 30571289 and 31071786) and National Indoor Conservation Center of Virus-free Germplasms of Fruit Crops for plant material collection.

Rights and permissions

Reprints and permissions

About this article

Cite this article

He, L.G., Wang, H.L., Liu, D.C. et al. Isolation and expression of a cold-responsive gene PtCBF in Poncirus trifoliata and isolation of citrus CBF promoters. Biol Plant 56, 484–492 (2012). https://doi.org/10.1007/s10535-012-0059-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10535-012-0059-5

Additional key words

Navigation