Plant Molecular Biology

, Volume 65, Issue 1, pp 13–30

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
Article

DOI: 10.1007/s11103-007-9190-7

Cite this article as:
Oudin, A., Mahroug, S., Courdavault, V. et al. Plant Mol Biol (2007) 65: 13. doi:10.1007/s11103-007-9190-7

Abstract

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.

Keywords

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

Abbreviations

DXS

1-Deoxy-d-xylulose 5-phosphate synthase

DXR

1-Deoxy-d-xylulose 5-phosphate reductoisomerase

ESMB

Early steps in monoterpenoid biosynthesis

G10H

Geraniol 10-hydroxylase

HDS

Hydroxymethylbutenyl 4-diphosphate synthase

HMBPP

Hydroxymethylbutenyl 4-diphosphate

IM

Inducing medium

IPAP

Internal phloem associated parenchyma

MECS

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

MeJa

Methyljasmonate

MEP

Methyl-d-erythritol 4-phosphate

MIA

Monoterpene indole alkaloid

MM

Maintenance medium

PM

Production medium

SLS

Secologanin synthase

STR

Strictosidine synthase

T16H

Tabersonine 16-hydroxylase

TEM

Transmission electron microscopy

Supplementary material

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