methyl-jasmonate and catharanthine

Long-term stable cell growth and production of vindoline, catharanthine, and ajmalicine of cambial meristematic cells (CMCs) from Catharanthus roseus were observed after 2 years of culture. C. roseus CMCs were treated with β-cyclodextrin (β-CD) and methyl jasmonate (MeJA) individually or in combination and were cultured both in conventional Erlenmeyer flasks (100, 250, and 500 mL) and in a 5-L stirred hybrid airlift bioreactor. CMCs of C. roseus cultured in the bioreactor showed higher yields of vindoline, catharanthine, and ajmalicine than those cultured in flasks. CMCs of C. roseus cultured in the bioreactor and treated with 10 mM β-CD and 150 μM MeJA gave the highest yields of vindoline (7.45 mg/L), catharanthine (1.76 mg/L), and ajmalicine (58.98 mg/L), concentrations that were 799, 654, and 426 % higher, respectively, than yields of CMCs cultured in 100-mL flasks without elicitors. Quantitative reverse transcription (RT)-PCR showed that β-CD and MeJA upregulated transcription levels of genes related to the biosynthesis of terpenoid indole alkaloids (TIAs). This is the first study to report that β-CD induced the generation of NO, which plays an important role in mediating the production of TIAs in C. roseus CMCs. These results suggest that β-CD and MeJA can enhance the production of TIAs in CMCs of C. roseus, and thus, CMCs of C. roseus have significant potential to be an industrial platform for production of bioactive alkaloids.

Topics: Acetates; beta-Cyclodextrins; Catharanthus; Cells, Cultured; Cyclopentanes; Oxylipins; Plant Cells; Secologanin Tryptamine Alkaloids; Vinblastine; Vinca Alkaloids

Catharanthus roseus (L.) G. Don (Apocynaceae) is a medicinal plant that produces more than 130 alkaloids, with special attention given to the production of the anti-hypertensive monomeric indole alkaloids, serpentine and ajmalicine, and the antitumor dimeric alkaloids, vinblastine and vincristine.. This study evaluated the cytotoxic activity of the indole alkaloid-enriched bioactive extract obtained from suspension cultured-cells of C. roseus elicited with methyl jasmonate (MJ) and cyclodextrins (CDs) in three cell lines: JURKAT E.6 human lymphocytic leukemia, THP-1 human monocytic leukemia and BL 1395 non-tumor human B-cell line.. An indole alkaloid-enriched bioactive extract was obtained from C. roseus cell cultures elicited with MJ and CDs. The indole alkaloids were identified using an HPLC-diode array system coupled to a time-of-flight mass spectrometer using electrospray ionization (ESI) source. The cytotoxic assays were made using the colorimetric assay 2, 3-bis (2-methoxy-4-nitro-5-sulfophenyl)-S-[(phenylamino)carbonyl]-2 tetrazolium hydroxide (XTT).. Four indole alkaloids were identified (catharanthine, ajmalicine, tabersonine and lochnericine) but only catharanthine and ajmalicine were quantified. The concentration of the indole alkaloid-enriched bioactive extract that inhibited cell growth by 50% was 211 and 210 ng/mL for the JURKAT E.6 and THP-1 cell lines, respectively.. The results confirm that the powerful antitumor activity of this indole alkaloid-enriched bioactive extract is not due to the effect of a single compound but depends on the synergistic action of the four compounds identified.

Topics: Acetates; Antineoplastic Agents, Phytogenic; Catharanthus; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cells, Cultured; Chromatography, High Pressure Liquid; Cyclodextrins; Cyclopentanes; Drug Discovery; Humans; Indole Alkaloids; Inhibitory Concentration 50; Oxylipins; Plant Extracts; Plant Leaves; Quinolines; Secologanin Tryptamine Alkaloids; Spectrometry, Mass, Electrospray Ionization; Vinca Alkaloids

Jasmonates and nitric oxide (NO) play important roles in the regulation of the signaling network leading to the biosynthesis of plant secondary metabolites. In this work, we explore the effect of constitutive overexpression of CrORCA3 (octadecanoid-responsive Catharanthus AP2/ERF domain), methyl jasmonate (MeJA), and sodium nitroprusside (SNP) on the differentiated tissue of Catharanthus roseus hairy roots. The changes in catharanthine concentration and in the levels of mRNA transcripts of pathway genes and regulators were tracked for 192 h. ORCA3 overexpression led to a slight decrease of the accumulation of catharanthine, while MeJA treatment caused a large increase in the levels of transcripts of pathway genes and the catharanthine concentration. SNP treatment alone or SNP in combination with MeJA treatment caused a dramatic decrease of the cathanranthine concentration, while at the same time the levels of transcripts of zinc finger-binding proteins genes (ZCTs) increased. The latter treatment also caused a decrease of the levels of transcripts of type-I protein prenyltransferase gene (PGGT-I). This response of transcriptional repressors and pathway genes may explain the antagonistic effects of NO and MeJA on catharanthine biosynthesis in C. roseus hairy roots.

Topics: Acetates; Biosynthetic Pathways; Catharanthus; Cyclopentanes; Gene Expression Profiling; Gene Expression Regulation, Plant; Genes, Regulator; Nitric Oxide; Oxylipins; Plant Proteins; Plant Roots; Tissue Culture Techniques; Transcription Factors; Vinca Alkaloids

The effects of methyl jasmonate, salicylic acid and ethylene on alkaloid accumulation in in vitro cell suspension, hairy roots and rootless shoot cultures of Catharanthus roseus were analyzed. Ajmalicine, but not catharanthine, accumulation was promoted by jasmonate and ethylene treatments in cell suspensions. In hairy roots, jasmonate induced the accumulation of both alkaloids, whereas ethylene only induced catharanthine accumulation. In shoot cultures, positive effects of jasmonate and ethylene were recorded only in vindoline accumulation. Ethylene diminished catharanthine accumulation in these cultures. No effect of salicylic acid was observed in any of the studied in vitro culture systems.

Topics: Acetates; Catharanthus; Cells, Cultured; Cyclopentanes; Ethylenes; Oxylipins; Plant Roots; Plant Shoots; Salicylic Acid; Secologanin Tryptamine Alkaloids; Vinca Alkaloids