Skip to main content
Log in

Downregulation of terpenoid indole alkaloid biosynthetic pathway by low temperature and cloning of a AP2 type C-repeat binding factor (CBF) from Catharanthus roseus (L). G. Don

  • Physiology and Biochemistry
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

Plants produce secondary metabolites in response to various external signals. Coordinated transcriptional control of biosynthetic genes emerges as a major mechanism dictating the accumulation of secondary metabolites in plant cells. However, information about stress regulation of secondary metabolites and the molecular mechanisms regulating these specialized pathways are poorly understood. Here, we show that terpenoid indole alkaloid (TIA) biosynthetic pathway is differentially regulated in response to different abiotic stresses in Catharanthus roseus, a model medicinal plant producing important anticancer and antihypertensive drugs. Semiquantitative RT-PCR analysis of TIA and related primary pathway genes in response to dehydration, low temperature, salinity, UV-light and wounding revealed their negative regulation in response to low temperature. HPLC analysis further supports the notion that TIA biosynthetic pathway is negatively controlled by low temperature stress. Furthermore, we report the cloning of a C-repeat binding transcription factor from C. roseus (CrCbf), belonging to AP2 class of transcription factor and possessed the NLS and CBF signature s equence characteristic of CBFs. CrCbf was found to be similar to Brassica Cbfs, whereas it was distant to monocot Cbfs. Southern analysis of CrCbf revealed the presence of more than one copy of CrCbf gene or other Cbf homologues in C. roseus genome. The transcription of CrCbf was found to be constitutive in response to low temperature but it showed differential distribution. The need for identifying novel transcription factors in understanding secondary metabolite biosynthesis is discussed.

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
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Abbreviations

AP2:

Apetala 2

CBF:

c-Repeat binding factor

ERF:

Ethylene responsive factor

HPLC:

High performance liquid chromatography

MeJA:

Methyl ester of jasmonic acid

MEP:

2-C-methyl-d-erythritol-4-phosphate

ORCA:

Octadecanoid-derivative responsive Catharanthus AP2-domain protein

RT-PCR:

Reverse transcriptase-polymerase chain reaction

TIA:

Terpenoid indole alkaloid

References

  • Altschul SF, Madden TL, Schaffe AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  PubMed  CAS  Google Scholar 

  • Batra J, Dutta A, Singh D, Kumar S, Sen J (2004) Growth and terpenoid indole alkaloid production in Catharanthus roseus hairy root clones in relation to left- and right-termini-linked Ri T-DNA gene integration. Plant Cell Rep 23:148–154

    Article  PubMed  CAS  Google Scholar 

  • Choi DW, Rodriguez EM, Close TJ (2002) Barley Cbf3 gene identification, expression pattern, and map location. Plant Physiol 129:1781–1788

    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 

  • Dutta A, Batra J, Pandey Rai S, Singh D, Kumar S, Sen J (2005) Expression of terpenoid indole alkaloid biosynthetic pathway genes corresponds to accumulation of related alkaloids in Catharanthus roseus (L.) G. Don Planta 220:376–383

    Article  PubMed  CAS  Google Scholar 

  • Dutta A, Singh D, Kumar S, Sen J (2007) Transcript profiling of terpenoid indole alkaloid pathway genes and regulators reveals strong expression of repressors in Catharanthus roseus cell cultures. Plant Cell Rep (in press)

  • Eisen MB, Spellman PT, Brown PO, Botstein D (1998) Cluster analysis and display of genome-wide expression patterns. Proc Natl Acad Sci USA 97:4985–4990

    Google Scholar 

  • Fukao T, Xu K, Ronald PC, Bailey-Serres J (2006) A variable cluster of ethylene response factor-like genes regulates metabolic and developmental acclimation responses to submergence in rice. Plant Cell 18:2021–2034

    Article  PubMed  CAS  Google Scholar 

  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158

    Article  PubMed  CAS  Google Scholar 

  • Gao MJ, Allard G, Byass L, Flanagan AM, Singh J (2002) Regulation and characterization of four CBF transcription factors from Brassica napus. Plant Mol Biol 49:459–471

    Article  PubMed  CAS  Google Scholar 

  • Jaglo KR, Kleff S, Amundsen KL, Zhang X, Haake V, Zhang JZ, Deits T, Thomashow MF (2001) 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

    Article  PubMed  CAS  Google Scholar 

  • Kumar S, Tamura K, Jakobsen IB, Nei M (2002) Molecular evolutionary genetics analysis software. Bioinform Appl Notes 17:1244–1245

    Google Scholar 

  • Marone M, Mozzetti S, De Ritis D, Pierelli L, Scambia G (2001) Semiquantitative RT-PCR analysis to assess the expression levels of multiple transcripts from the same sample. Biol Proced Online 3:19–25

    Article  PubMed  CAS  Google Scholar 

  • Menke FL, Parchmann S, Mueller MJ, Kijne JW, Memelink J (1999a) Involvement of the octadecanoid pathway and protein phosphorylation in fungal elicitor-induced expression of terpenoid indole alkaloid biosynthetic genes in Catharanthus roseus. Plant Physiol 119:1289–1296

    Article  PubMed  CAS  Google Scholar 

  • Menke FL, Champion A, Kijne JW, Memelink J (1999b) A novel jasmonate- and elicitor-responsive element in the periwinkle secondary metabolite biosynthetic gene Str interacts with a jasmonate- and elicitor-inducible AP2-domain transcription factor, ORCA2. EMBO J 18:4455–4463

    Article  PubMed  CAS  Google Scholar 

  • Nimchuk Z, Eulgem T, Holt III BF, Dangl JF (2003) Recognition and response in the plant immune system. Annu Rev Genet 37:579–609

    Article  PubMed  CAS  Google Scholar 

  • Ouwerkerk PB, Hallard D, Verpoorte R, Memelink J (1999) Identification of UV-B light-responsive regions in the promoter of the tryptophan decarboxylase gene from Catharanthus roseus. Plant Mol Biol 41: 491–503

    Article  PubMed  CAS  Google Scholar 

  • Pasquali G, Goddijn OJ, de Waal A, Verpoorte R, Schilperoort RA, Hoge JH, Memelink J (1992) Coordinated regulation of two indole alkaloid biosynthetic genes from Catharanthus roseus by auxin and elicitors. Plant Mol Biol 18:1121–1131

    Article  PubMed  CAS  Google Scholar 

  • Pauw B, Hilliou FA, Martin VS, Chatel G, de Wolf CJ, Champion A, Pre M, van Duijn B, Kijne JW, van der Fits, Memelink J (2004) Zinc finger proteins act as transcriptional repressors of alkaloid biosynthesis genes in Catharanthus roseus. J.Biol Chem 279:52940–52948

    Article  PubMed  CAS  Google Scholar 

  • Reichmann JL, Mayerowitz EM (1998) The AP2/EREBP family of transcription factors. J Biol Chem 379:633–649

    Article  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 

  • Siberil Y, Benhamron S, Memelink J, Giglioli-Guivarc’h N, Thiersault M, Boisson B, Doireau P, Gantet P (2001) Catharanthus roseus G-box binding factors 1 and 2 act as repressors of strictosidine synthase gene expression in cell cultures. Plant Mol Biol 45:477–488

    Article  PubMed  CAS  Google Scholar 

  • Stockinger EJ, Gilmour SJ, Thomashow MF (1997) Arabidopsis thaliana CBF1 encodes an AP2 domain—containing transcription activator that bind to the C-repeats/ DRE, a cis-acting DNA regulatory elements that stimulate transcription in response to low temperature and water deficit. Proc Natl Acad Sci USA 94:1035–1040

    Article  PubMed  CAS  Google Scholar 

  • Thomashow MF (2001) So what’s new in the field of plant cold acclimation? Lots! Plant Physiol 125:89–93

    Article  PubMed  CAS  Google Scholar 

  • van der Fits L, Memelink J (2000) ORCA3, a jasmonate-responsive transcriptional regulator of plant primary and secondary metabolism. Science 289:295–297

    Article  PubMed  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Zhang X, Fowler SG, Cheng H, Lou Y, Rhee SY, Stockinger EJ, Thomashow MF (2004) 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

    Article  PubMed  CAS  Google Scholar 

  • Zhao T-J, Sun S, Liu Y, Liu JM, Liu Q, Yan YB, Zhou HM (2006) Regulating the drought-responsive element (DRE)-mediated signaling pathway by synergic functions of trans-active and trans-inactive DRE binding factors in Brassica napus. J Biol Chem 281:10752–10759

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgement

AD acknowledges Council of Scientific and Industrial Research (CSIR), Government of India for the award of senior research fellowship. This work was supported by the Department of Biotechnology, Government of India. AD pay her tribute and dedicate this paper to Dr Jayanti Sen, who unfortunately passed away before this work was published.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Renu Deswal.

Additional information

Communicated by P. Lakshmanan.

This paper is dedicated to inspirational memory of Dr. Jayanti Sen. The cDNA sequence of CrCbf have been submitted to Genbank under the accession number DQ989629.

Electronic supplementary material

Below is the link to the electronic supplementary material.

(DOC 49 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dutta, A., Sen, J. & Deswal, R. Downregulation of terpenoid indole alkaloid biosynthetic pathway by low temperature and cloning of a AP2 type C-repeat binding factor (CBF) from Catharanthus roseus (L). G. Don. Plant Cell Rep 26, 1869–1878 (2007). https://doi.org/10.1007/s00299-007-0383-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-007-0383-y

Keywords

Navigation