Enhanced Alkaloid Production from Cell Culture System of Catharanthus roseus in Combined Effect of Nutrient Salts, Sucrose and Plant Growth Regulators

Malay Ranjan Mishra,Rajesh Kumar Srivastava,Nasim Akhtar,Hammad Afzal

doi:10.14302/issn.2576-6694.jbbs-18-2475

Malay Ranjan Mishra, Rajesh Kumar Srivastava + Show 2 more

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https://doi.org/10.14302/issn.2576-6694.jbbs-18-2475

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Abstract

Callus and biomass culture of Catharanthus roseus L. were established to check for the presence of total alkaloid and its subsequent yield. Various treatments like strength of nutrient salts, sucrose concentrations and combinations of plant growth regulators (PGR’s) were applied to both MS and B5 in agar as well as suspension medium to test their effects on enhanced alkaloid content and its yield. There was no significant difference in any of the observable parameters of fresh wt, dry wt, alkaloid content, production, productivity and yield if the culture were treated similarly in both types of media formulations (MS or B5 salts). Physical state (agar solidified or the liquid suspension) of the medium had significant effect on all the parameters particular on fresh wt, alkaloid content and yields. Although, the fresh wt. and dry wt. of biomass in suspension culture was 2-3 times less than that of callus obtained from agar medium. However, the alkaloid content and yield was 2-3 times higher in suspension culture compared to agar medium in similar treatments. The highest alkaloid content observed was 5.67mg/g dwt in B5 suspension medium containing 3% sucrose and modified with 0.5mg/l 2,4-Dichlorophenoxy acetic acid (2,4-D) + 1 mg/l Kinetin (KIN) + 2mg/l α- naphthalene acetic acid (NAA). The combined effects of these factors on the enhanced production of total alkaloids were expected to contain higher yield of anticancer vinblastine and vincristine in the cell suspension culture system.

Highlights

Plants are rich source of a number of secondary metabolites including the alkaloids with an estimated count of around 3000 different types till date [1]
These indole alkaloids such as antimalarial quinine from Chincona officinalis, the antineoplastic camptothecine from Camptotheca acuminta, the rat poison and tonic like strychnine from Strychnos nux-vomica, antihypertensive and tranquilizer resperine from Rauvolfia sp., and the widely investigated antitumor agents vinblastine and vincristine from Catharanthus roseus are being prescribed in modern therapeutics to treat the diseases
The standard culture conditions induced considerable amount of variations in total alkaloid production from callus grown on agar medium and the cell biomass in suspension of Catharanthus roseus

Summary

Introduction

Plants are rich source of a number of secondary metabolites including the alkaloids with an estimated count of around 3000 different types till date [1]. TIA pathway depends on indole and terpenoid precursors supplied by two convergent branches of the primary metabolites the shikimate and the isopentenyl diphosphate pathways It involves more than twenty enzymes located in cytosol, (pro)vacuoles and chloroplasts/plastids [4]. (Apocynacea) known as Madagascar periwinkle is one of the most extensively used and investigated medicinal plant [5] This is a major source of the highly important anti-cancer bisindole alklaoids i.e. vinblastine and vincristine [6,7,8,9]. Antihypertensive alkaloids like ajmalicine and serpentine are found in the roots of these plants [11,12] Apart form these activities this plant is found to be anti-microbial where extracts of the plant is found to be effective against S. typhi, S. aureus and B. subtilis [13,14]. Owing to very trace amount in C. roseus, has initiated widespread search for finding alternative solutions for cost effective and mass production of these drugs [7] [27,28]

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