Production of Alkaloids in Plant Cell Culture

Roberts, Margaret

doi:10.1007/978-1-4757-2905-4_7

Cited by 4 publications

(1 citation statement)

References 103 publications

(87 reference statements)

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“…The importance of the vacuole also implies that cell proliferation and alkaloid production are negatively correlated. In fact, the production of alkaloids is higher during the stationary phase (Roberts 1998). In the current study, we reinforced the transition from proliferation to cell expansion by auxin depletion and were able to promote catharanthine accumulation in strain C4 almost 20-fold.…”

Section: Discussionmentioning

confidence: 99%

Cell Type Matters: Competence For Alkaloid Metabolism Differs In Two Seed-Derived Cell Strains of Catharanthus Roseus.

Raorane

Manz

Hildebrandt

et al. 2021

Preprint

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Since the discovery of the anticancer drugs vinblastine and vincristine, Catharanthus roseus has been intensively studied for biosynthesis of several terpene indole alkaloids (TIAs). Due to their low abundance in plant tissues at a simultaneously high demand, modes of production alternative to conventional extraction are mandatory. Plant cell fermentation might become one of these alternatives, yet decades of research have shown limited success to certain product classes, leading to the question, how to preserve the intrinsic ability to produce TIAs (metabolic competence) in cell culture? We used the strategy to use the developmental potency of mature embryos to generate such strains. Two cell strains (C1and C4) from seeds of Catharanthus roseus were found not only to differ morphologically, but also in their metabolic competence. This differential competence became manifest under phytohormone elicitation, but also upon feeding with alkaloid pathway precursors. The more active strain C4 formed larger cell aggregates and was endowed with longer mitochondria. These cellular features were accompanied by higher alkaloid accumulation in response to methyl jasmonate (MeJA) elicitation. The levels of catharanthine could be increased significantly, while the concurrent vindoline branch of the pathway was blocked, such that no bisindole alkaloids were detectable. By feeding vindoline to MeJA elicited C4 cells, vincristine became detectable; however, only to marginal amounts. In conclusion, these results show that cultured cells are not just “de-differentiated”, but can differ in metabolic competence. In addition to elicitation, and precursor feeding, the cellular properties of the “biomatter” are highly relevant for the success of plant cell fermentation.

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Section: Discussionmentioning

confidence: 99%

Cell Type Matters: Competence For Alkaloid Metabolism Differs In Two Seed-Derived Cell Strains of Catharanthus Roseus.

Raorane

Manz

Hildebrandt

et al. 2021

Preprint

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Production of Pharmaceuticals from Papaver Cultivars in Vitro

Tisserat¹,

Berhow²

2009

Engineering in Life Sciences

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Iranian (Papaver bracteatum Lindl.) and opium poppy (P. somniferum L.) plantlets obtained from germinated seeds grown on a Murashige and Skoog basal medium (BM) readily manifest alkaloids. Temperature had a profound effect on growth and alkaloid production after 8 weeks in culture. Plantlets of poppy cultivars (cvs.) grew best at 18.5 and 20°C compared to 15 or 25°C. An alkaloid survey study with 24 Iranian and 21 opium poppy cvs. revealed that total morphinan alkaloids ranged from 0 to 6.55 mg/g dw. Prolific axillary branching was achieved from poppy cvs. by maintaining shoots on BM containing 1.0 mg/L N6‐benzyladenine and 0.01 mg/L α‐naphthalene acetic acid for an additional 16 weeks. The influence of vessel size on the growth response of established shoot clumps was determined by subculture in a variety of culture vessels for 8 weeks. The tested culture vessels included culture tubes (55 mm3 capacity (cap.)), babyfood jars (143 mm3 cap.), Magenta GA‐7 containers (365 mm3 cap.), and polycarbonate jars (1890 mm3 cap.) employing an in vitro hydroponics system (i.e. an automated plant culture system (APCS)). Highest growth rates occurred employing the APCS. The culture vessel capacity had a significant positive correlation on shoot length, fresh weight, number of leaves, and number of shoots. Shoot length, fresh weight, leaves, and shoots grown in the APCS exhibited increases of 1‐, 21.5‐, 7.8‐, and 8.3‐fold, respectively, compared to shoots grown in culture tubes. Higher culture growth rates that occurred in the larger‐size vessels were correlated with lower alkaloid production (mg alkaloids/g dw). However, the overall total alkaloids/vessel [(mg alkaloid/g dw)×g culture dw] increased because of greater biomass production per vessel. The alkaloid content was found to remain stable for shoots grown over a 6–month evaluation period.

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Cell type matters: competence for alkaloid metabolism differs in two seed-derived cell strains of Catharanthus roseus

et al. 2022

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Since the discovery of the anticancer drugs vinblastine and vincristine, Catharanthus roseus has been intensively studied for biosynthesis of several terpene indole alkaloids (TIAs). Due to their low abundance in plant tissues at a simultaneously high demand, modes of production alternative to conventional extraction are mandatory. Plant cell fermentation might become one of these alternatives, yet decades of research have shown limited success to certain product classes, leading to the question: how to preserve the intrinsic ability to produce TIAs (metabolic competence) in cell culture? We used the strategy to use the developmental potency of mature embryos to generate such strains. Two cell strains (C1and C4) from seed embryos of Catharanthus roseus were found to differ not only morphologically, but also in their metabolic competence. This differential competence became manifest not only under phytohormone elicitation, but also upon feeding with alkaloid pathway precursors. The more active strain C4 formed larger cell aggregates and was endowed with longer mitochondria. These cellular features were accompanied by higher alkaloid accumulation in response to methyl jasmonate (MeJA) elicitation. The levels of catharanthine could be increased significantly, while the concurrent vindoline branch of the pathway was blocked, such that no bisindole alkaloids were detectable. By feeding vindoline to MeJA-elicited C4 cells, vincristine became detectable; however, only to marginal amounts. In conclusion, these results show that cultured cells are not “de-differentiated”, but can differ in metabolic competence. In addition to elicitation and precursor feeding, the cellular properties of the “biomatter” are highly relevant for the success of plant cell fermentation.

show abstract

Production of Alkaloids in Plant Cell Culture

Cited by 4 publications

References 103 publications

Cell Type Matters: Competence For Alkaloid Metabolism Differs In Two Seed-Derived Cell Strains of Catharanthus Roseus.

Cell Type Matters: Competence For Alkaloid Metabolism Differs In Two Seed-Derived Cell Strains of Catharanthus Roseus.

Production of Pharmaceuticals from Papaver Cultivars in Vitro

Cell type matters: competence for alkaloid metabolism differs in two seed-derived cell strains of Catharanthus roseus

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