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Epidermis is a pivotal site of at least four secondary metabolic pathways in Catharanthus roseus aerial organs

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Abstract

Catharanthus roseus produces a wide range of secondary metabolites, some of which present high therapeutic values such as antitumoral monoterpenoid indole alkaloids (MIAs), vinblastine and vincristine, and the hypotensive MIA, ajmalicine. We have recently shown that a complex multicellular organisation of the MIA biosynthetic pathway occurred in C. roseus aerial organs. In particular, the final steps of both the secoiridoid–monoterpene and indole pathways specifically occurred in the epidermis of leaves and petals. Chorismate is the common precursor of indole and phenylpropanoid pathways. In an attempt to better map the spatio-temporal organisation of diverse secondary metabolisms in Catharanthus roseus aerial organs, we studied the expression pattern of genes encoding enzymes of the phenylpropanoid pathway (phenylalanine ammonia-lyase [PAL, E.C. 4.3.1.5], cinnamate 4-hydroxylase [C4H, E.C. 1.14.13.11] and chalcone synthase [CHS, E.C. 2.3.1.74]). In situ hybridisation experiments revealed that CrPAL and CrC4H were specifically localised to lignifying xylem, whereas CrPAL, CrC4H and CrCHS were specifically expressed in the flavonoid-rich upper epidermis. Interestingly, these three genes were co-expressed in the epidermis (at least the upper, adaxial one) together with three MIA-related genes, indicating that single epidermis cells were capable of concomitantly producing a wide range of diverse secondary metabolites (e.g. flavonoïds, indoles, secoiridoid–monoterpenes and MIAs). These results, and data showing co-accumulation of flavonoids and alkaloids in single cells of C. roseus cell lines, indicated the spatio-temporal feasibility of putative common regulation mechanisms for the expression of these genes involved in at least four distinct secondary metabolisms.

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Abbreviations

MIA(s):

Monoterpenoid indole alkaloid(s)

PAL:

Phenylalanine ammonia-lyase

C4H:

Cinnamate 4-hydroxylase

CHS:

Chalcone synthase

MEP:

2C-methyl-d-erythritol 4-phosphate

DXS:

1-deoxy-d-xylulose 5-phosphate (DXP) synthase

DXR:

DXP reductoisomerase

MECS:

2C-methyl-d-erythritol 2,4-cyclodiphosphate (MEC) synthase

G10H:

Geraniol 10-hydroxylase

SLS:

Secologanin synthase

TDC:

Tryptophan decarboxylase

STR:

Strictosidine synthase

D4H:

Desacetoxyvindoline 4-hydroxylase

DAT:

Deacetylvindoline 4-O-acetyltransferase

2,4-D:

2,4-dichlorophenoxyacetic acid

MM:

Maintenance medium

PM:

Production medium

FAA:

Formaldehyde acetic acid ethyl alcohol

2-APB:

2-aminoethyldiphenyl borinate

AP:

Alkaline phosphatase

BCIP:

5-Bromo-4-chloro-3-indolyl phosphate

NBT:

Nitro blue tetrazolium chloride

ORCA:

Octadecanoid-responsive Catharanthus AP2 transcription factor

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Acknowledgements

This research was financially supported by the Ministère de l’Education Nationale, de la Recherche et de la Technologie (MENRT, France) and by the Ligue Nationale contre le Cancer (comité d’indre and comité d’indre et loire). S. M. was financed by a “bourse de cooperation franco-algérienne”. We thank Dr. Kiyota (National Institute of Agrobiological Sciences, Tsukuba Ibaraki, Japan) for the gift of C. roseus PAL cDNA and Prof. J. Schröder (University of Freiburg, Germany) who kindly provided C. roseus C4H and CHS cDNAs. We also thank Dr. Facchini (University of Calgary, Canada) for careful revision of the manuscript.

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Authors and Affiliations

  1. Université François-Rabelais de Tours, EA 2106 “Biomolécules et Biotechnologies Végétales” UFR Sciences et Techniques, Parc de Grandmont, 37200, Tours, France

    Samira Mahroug, Vincent Courdavault, Martine Thiersault, Benoit St-Pierre & Vincent Burlat

  2. Université François-Rabelais de Tours, Unité sous Contrat reconnue par l’INRA “Facteurs de transcription et ingénierie métabolique végétale”, Tours, France

    Samira Mahroug, Martine Thiersault, Benoit St-Pierre & Vincent Burlat

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  1. Samira Mahroug
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  2. Vincent Courdavault
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Correspondence to Vincent Burlat.

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Mahroug, S., Courdavault, V., Thiersault, M. et al. Epidermis is a pivotal site of at least four secondary metabolic pathways in Catharanthus roseus aerial organs. Planta 223, 1191–1200 (2006). https://doi.org/10.1007/s00425-005-0167-y

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