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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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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DOI: https://doi.org/10.1007/s00425-005-0167-y