Plant Molecular Biology

, Volume 65, Issue 1–2, pp 13–30 | Cite as

Spatial distribution and hormonal regulation of gene products from methyl erythritol phosphate and monoterpene-secoiridoid pathways in Catharanthus roseus

  • Audrey Oudin
  • Samira Mahroug
  • Vincent Courdavault
  • Nadège Hervouet
  • Charles Zelwer
  • Manuel Rodríguez-Concepción
  • Benoit St-Pierre
  • Vincent Burlat


The monoterpene indole alkaloids (MIAs) from Madagascar periwinkle (Catharanthus roseus) are secondary metabolites of high interest due to their therapeutical values. Secologanin, the monoterpenoid moiety incorporated into MIAs, is derived from the plastidial methyl-d-erythritol 4-phosphate (MEP) pathway. Here, we have cloned a cDNA encoding hydroxymethylbutenyl diphosphate synthase (HDS), a MEP pathway enzyme, and generated antibodies to investigate the distribution of transcripts and protein in MIA-producing aerial tissues. Consistent with our earlier work, transcripts for the genes encoding the so-called early steps in monoterpenoid biosynthesis (ESMB) enzymes (HDS, others MEP pathway enzymes and geraniol 10-hydroxylase) were preferentially co-localized to internal phloem associated parenchyma (IPAP) cells. By contrast, transcripts for the enzyme catalysing the last biosynthetic step to secologanin, secologanin synthase, were found in the epidermis. A coordinated response of ESMB genes was also observed in cell cultures stimulated to synthesise MIAs by hormone treatment, whereas no changes in SLS expression were detected under the same experimental conditions. Immunocytolabelling studies with the HDS-specific serum demonstrated the localisation of HDS to the plastid stroma and revealed that HDS proteins were most abundant in IPAP cells but could also be found in other cell types, including epidermal and mesophyll cells. Besides showing the existence of post-transcriptional mechanisms regulating the levels of HDS in C. roseus cells, our results support that intercellular translocation likely plays an important role during monoterpene-secoiridoid assembly.


Catharanthus roseus Compartmentation Coordinated regulation Hydroxymethylbutenyl 4-diphosphate synthase Methyl erythritol phosphate pathway Monoterpene indole alkaloids 



1-Deoxy-d-xylulose 5-phosphate synthase


1-Deoxy-d-xylulose 5-phosphate reductoisomerase


Early steps in monoterpenoid biosynthesis


Geraniol 10-hydroxylase


Hydroxymethylbutenyl 4-diphosphate synthase


Hydroxymethylbutenyl 4-diphosphate


Inducing medium


Internal phloem associated parenchyma


2C-methyl-d-erythritol 2,4-diphosphate synthase




Methyl-d-erythritol 4-phosphate


Monoterpene indole alkaloid


Maintenance medium


Production medium


Secologanin synthase


Strictosidine synthase


Tabersonine 16-hydroxylase


Transmission electron microscopy



This research was financially supported by the Ministère de l’Education Nationale, de la Recherche et de la Technologie (MENRT, France), by Biotechnocentre, by the Conseil Régional du Centre, by the Ligue contre le Cancer (comité d’Indre et Loire and comité de l’Indre) and by the Spanish Ministerio de Educacion y Ciencia and FEDER (grant BIO2005–00367 to MR-C). We thank Dr. J. Memelink (University of Leiden, the Netherlands) who kindly provided C. roseus G10H cDNA and the oriented cDNA library, and Dr. N. Campos (Universitat de Barcelona) for the EcAB3-3 strain. We also thank Dr. B. Arbeille and the staff of the Electron Microscopy Platform (CHU Bretonneau, Tours, France) for providing excellent working conditions for ultramicrotomy and TEM studies.

Supplementary material


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Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Audrey Oudin
    • 1
  • Samira Mahroug
    • 2
    • 5
  • Vincent Courdavault
    • 2
  • Nadège Hervouet
    • 3
  • Charles Zelwer
    • 3
  • Manuel Rodríguez-Concepción
    • 4
    • 6
  • Benoit St-Pierre
    • 2
    • 5
  • Vincent Burlat
    • 2
    • 5
  1. 1.EA 2106 “Biomolécules et Biotechnologies Végétales”, UFR des Sciences PharmaceutiquesUniversité François Rabelais de ToursToursFrance
  2. 2.EA 2106 “Biomolécules et Biotechnologies Végétales”, UFR des Sciences et TechniquesUniversité François Rabelais de ToursToursFrance
  3. 3.Centre de Biophysique MoléculaireUPR 4301, CNRSOrleans cedex 02France
  4. 4.Departament de Bioquímica i Biologia Molecular, Facultat de BiologiaUniversitat de BarcelonaBarcelonaSpain
  5. 5.Unité sous Contrat reconnue par l’INRA “Facteurs de transcription et ingénierie métabolique végétale”Université François Rabelais de ToursToursFrance
  6. 6.Consorci CSIC-IRTA de Genetica Molecular VegetalBarcelonaSpain

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