Spatial organization of the vindoline biosynthetic pathway in Catharanthus roseus

Guirimand, Grégory; Guihur, Anthony; Poutrain, Pierre; Héricourt, François; Mahroug, Samira; St-Pierre, Benoit; Burlat, Vincent; Courdavault, Vincent

doi:10.1016/j.jplph.2010.08.018

Cited by 79 publications

(74 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…5,E and F). These results are consistent with the occurrence of four CYP71D351 ESTs in the C. roseus EST epidermome database that were previously incorrectly annotated as CYP71D12 (Supplemental Table S1; Murata et al, 2008) and are also in agreement with the postulated localization of the first two steps of vindoline biosynthesis in the epidermis (Murata and De Luca, 2005;Murata et al, 2008;Guirimand et al, 2011b).…”

Section: Cyp71d351 (T16h2) Transcripts Are Localized In the Epidermissupporting

confidence: 90%

“…FJ647194) could be efficiency amplified by PCR using complementary DNAs (cDNAs) prepared from C. roseus cell cultures (Guirimand et al, 2011b), all our attempts to carry out this amplification using reverse-transcribed RNAs extracted from young leaves failed. As a consequence, additional amplifications of a partial internal sequence were attempted using young leaf cDNAs and the previously described CrT16H-RT01 and CrRT16H-RT02 primers (Murata and De Luca, 2005).…”

Section: Isolation Of Cyp71d351 Complementary Dnamentioning

confidence: 99%

“…While transcripts and enzyme activities of both D4H and DAT have been localized to laticifers/idioblasts, adaxial and abaxial epidermal cells have been shown to host the starting steps of the biosynthetic pathway. The epidermal localization of 16OMT transcripts was first established using epidermis-specific mRNA enrichment procedures performed by carborundum abrasion or epidermis laser-capture microdissection/EST sequencing (Murata and De Luca, 2005;Levac et al, 2008;Murata et al, 2008) and was further confirmed by RNA in situ hybridization (Guirimand et al, 2011b). By contrast, the situation is less clear for T16H, whose only known coding sequence, CYP71D12, was cloned from C. roseus cell cultures (Schröder et al, 1999;Guirimand et al, 2011b).…”

mentioning

confidence: 99%

See 2 more Smart Citations

A Pair of Tabersonine 16-Hydroxylases Initiates the Synthesis of Vindoline in an Organ-Dependent Manner inCatharanthus roseus

et al. 2013

Self Cite

View full text Add to dashboard Cite

Hydroxylation of tabersonine at the C-16 position, catalyzed by tabersonine 16-hydroxylase (T16H), initiates the synthesis of vindoline that constitutes the main alkaloid accumulated in leaves of Catharanthus roseus. Over the last decade, this reaction has been associated with CYP71D12 cloned from undifferentiated C. roseus cells. In this study, we isolated a second cytochrome P450 (CYP71D351) displaying T16H activity. Biochemical characterization demonstrated that CYP71D12 and CYP71D351 both exhibit high affinity for tabersonine and narrow substrate specificity, making of T16H, to our knowledge, the first alkaloid biosynthetic enzyme displaying two isoforms encoded by distinct genes characterized to date in C. roseus. However, both genes dramatically diverge in transcript distribution in planta. While CYP71D12 (T16H1) expression is restricted to flowers and undifferentiated cells, the CYP71D351 (T16H2) expression profile is similar to the other vindoline biosynthetic genes reaching a maximum in young leaves. Moreover, transcript localization by carborundum abrasion and RNA in situ hybridization demonstrated that CYP71D351 messenger RNAs are specifically located to leaf epidermis, which also hosts the next step of vindoline biosynthesis. Comparison of high-and low-vindoline-accumulating C. roseus cultivars also highlights the direct correlation between CYP71D351 transcript and vindoline levels. In addition, CYP71D351 down-regulation mediated by virus-induced gene silencing reduces vindoline accumulation in leaves and redirects the biosynthetic flux toward the production of unmodified alkaloids at the C-16 position. All these data demonstrate that tabersonine 16-hydroxylation is orchestrated in an organ-dependent manner by two genes including CYP71D351, which encodes the specific T16H isoform acting in the foliar vindoline biosynthesis.

show abstract

Section: Cyp71d351 (T16h2) Transcripts Are Localized In the Epidermissupporting

confidence: 90%

Section: Isolation Of Cyp71d351 Complementary Dnamentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

A Pair of Tabersonine 16-Hydroxylases Initiates the Synthesis of Vindoline in an Organ-Dependent Manner inCatharanthus roseus

et al. 2013

Self Cite

View full text Add to dashboard Cite

show abstract

“…The great pharmacological importance of TIAs, associated with their low abundance in the plant (approximately 0.0005% dry weight), stimulated intense research on the TIA pathway, and C. roseus has become one of the most studied medicinal plants (van der Heijden et al, 2004;Verpoorte et al, 2007;Costa et al, 2008). TIA biosynthesis shows multicellular compartmentation in C. roseus leaves, with early steps occurring in epidermal cells and late steps occurring in laticifer and mesophyll idioblast cells, predicting intercellular translocation of TIA intermediates (St-Pierre et al, 1999;Murata et al 2008;Guirimand et al, 2011). Moreover, the TIA pathway also shows a complex subcellular organization, with different parts of the pathway being localized in the plastids, the vacuole, the cytosol, and the endoplasmic reticulum (ER), predicting further transport events (Mahroug et al, 2007;Guirimand et al, 2011).…”

mentioning

confidence: 99%

“…TIA biosynthesis shows multicellular compartmentation in C. roseus leaves, with early steps occurring in epidermal cells and late steps occurring in laticifer and mesophyll idioblast cells, predicting intercellular translocation of TIA intermediates (St-Pierre et al, 1999;Murata et al 2008;Guirimand et al, 2011). Moreover, the TIA pathway also shows a complex subcellular organization, with different parts of the pathway being localized in the plastids, the vacuole, the cytosol, and the endoplasmic reticulum (ER), predicting further transport events (Mahroug et al, 2007;Guirimand et al, 2011). In spite of the importance of all those TIA * Corresponding author; e-mail msottoma@ibmc.up.pt.…”

mentioning

confidence: 99%

Vacuolar Transport of the Medicinal Alkaloids from Catharanthus roseus Is Mediated by a Proton-Driven Antiport

et al. 2013

View full text Add to dashboard Cite

Catharanthus roseus is one of the most studied medicinal plants due to the interest in their dimeric terpenoid indole alkaloids (TIAs) vinblastine and vincristine, which are used in cancer chemotherapy. These TIAs are produced in very low levels in the leaves of the plant from the monomeric precursors vindoline and catharanthine and, although TIA biosynthesis is reasonably well understood, much less is known about TIA membrane transport mechanisms. However, such knowledge is extremely important to understand TIA metabolic fluxes and to develop strategies aimed at increasing TIA production. In this study, the vacuolar transport mechanism of the main TIAs accumulated in C. roseus leaves, vindoline, catharanthine, and a-39,49-anhydrovinblastine, was characterized using a tonoplast vesicle system. Vindoline uptake was ATP dependent, and this transport activity was strongly inhibited by NH 4 + and carbonyl cyanide m-chlorophenyl hydrazine and was insensitive to the ATP-binding cassette (ABC) transporter inhibitor vanadate. Spectrofluorimetry assays with a pH-sensitive fluorescent probe showed that vindoline and other TIAs indeed were able to dissipate an H + gradient preestablished across the tonoplast by either vacuolar H + -ATPase or vacuolar H + -pyrophosphatase. The initial rates of H + gradient dissipation followed Michaelis-Menten kinetics, suggesting the involvement of mediated transport, and this activity was species and alkaloid specific. Altogether, our results strongly support that TIAs are actively taken up by C. roseus mesophyll vacuoles through a specific H + antiport system and not by an ion-trap mechanism or ABC transporters.

show abstract

Structure and Synthesis of Vindolicine and Derivatives

Asia,

Sammer,

Vendier

et al. 2024

Chemistry & Biodiversity

View full text Add to dashboard Cite

This article describes the reaction of vindoline with formaldehyde and trimethyl orthoformate to prepare vindolicine, tris‐vindolicinyl methane and higher molecular weight homologues. The synthesis of 10‐formyl vindoline as an intermediate allowed further exploration of its chemistry, in particular the reaction with acetone which yielded a symmetrical dimer, which was further reacted with vindoline to give molecules containing three and four vindoline units. These molecules were characterized by NMR and for some of them (vindolicine, 10‐formyl vindoline, 10‐(1’‐(but‐1’‐en‐3’‐one))‐vindoline) by X‐ray crystallography. Depending on the substitution and on the absence of axes of symmetry, the NMR spectra displayed non‐equivalent spin systems for the vindoline moieties. The dimer formed from the double condensation of 10‐formyl vindoline with acetone showed cytotoxic activity in the micromolar range.

show abstract

Spatial organization of the vindoline biosynthetic pathway in Catharanthus roseus

Cited by 79 publications

References 33 publications

A Pair of Tabersonine 16-Hydroxylases Initiates the Synthesis of Vindoline in an Organ-Dependent Manner inCatharanthus roseus

A Pair of Tabersonine 16-Hydroxylases Initiates the Synthesis of Vindoline in an Organ-Dependent Manner inCatharanthus roseus

Vacuolar Transport of the Medicinal Alkaloids from Catharanthus roseus Is Mediated by a Proton-Driven Antiport

Structure and Synthesis of Vindolicine and Derivatives

Contact Info

Product

Resources

About