Development of a Cell Suspension Culture System for Promoting Alkaloid and Vinca Alkaloid Biosynthesis Using Endophytic Fungi Isolated from Local Catharanthus roseus

Abstract

Cell and tissue cultures of Catharanthus roseus have been studied extensively as an alternative strategy to improve the production of valuable secondary metabolites. The purpose of this study was to produce C. roseus callus and suspension cell biomass of good quality and quantity to improve the total alkaloids and bis-indole alkaloids. The young stem derived-callus of C. roseus variety Quang Ninh (QN) was grown on MS medium supplemented with 1.5 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) plus 1.5 mg/L kinetin, and the growth rate increased by 67-fold after 20 days. The optimal conditions for maintaining the cell suspension culture were 150 mg/50 mL cell inoculum, a medium pH of 5.5 and a culture temperature of 25 °C. The low alkaloid content in the culture was compensated for by using endophytic fungi isolated from local C. roseus. Cell extracts of endophytic fungi—identified as Fusarium solani RN1 and Chaetomium funicola RN3—were found to significantly promote alkaloid accumulation. This elicitation also stimulated the accumulation of a tested bis-indole alkaloid, vinblastine. The findings are important for investigating the effects of fungal elicitors on the biosynthesis of vinblastine and vincristine, as well as other terpenoid indole alkaloids (TIAs), in C. roseus QN cell suspension cultures.

Biosynthesis of anticancer phytochemical compounds and their chemistry

“…Cell components from related endophytic fungi may act as elicitors and increase the levels of secondary metabolites obtained from in-vitro cultures (Farhadi et al, 2020). Using endophytic fungal cell extracts from Catharanthus roseus plants such as Fusarium solani RN1 and Chaetomium funicola RN3 and using those extracts as an elicitor in suspension culture augmented the yield of both vincristine and vinblastine plant Frontiers in Pharmacology frontiersin.org alkaloid (Linh et al, 2021). Biotic elicitors, such as cell extract from endophytic fungi such as Alternaria sesami, were also utilized to boost the levels of alkaloids in the cell suspension culture of Catharanthus roseus (Birat et al, 2022).…”

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“…Colchicum autumnale L.) [20][21] Vincaalkaloids (isolated for Catharanthus roseus G. Don) 22 , Combretastatin A-4 phosphate (isolated from Combretum caffrum) [23][24] and Noscapine (from Papaver somniferum) 25 prevent polymerization, whereas Paclitaxel (from Taxus brevifolia Nutt.) prevents dissociation by stabilizing the structure 26 Fig.…”

Abiotic Stresses Elicitation Potentiates the Productiveness of Cardoon Calli as Bio-Factories for Specialized Metabolites Production

“…The efficacy of elicitation for a specific target compound depends on the plant species, genotype, and tissue, and not least by the growth conditions (e.g., temperature, medium composition, and light) [ 7 ]. As an example, elicitation of Catharanthus roseus cell cultures with extracts of endophytic fungi allowed production of vinblastine and vincristine, an important class of tropane indol alkaloids, which was not previously been achieved in cultured cells [ 8 ]. Similarly, undifferentiated callus tissues of Cannabis sativa , even those derived from flowers, were not able to synthesize cannabinoids, unless methyl jasmonate was added to the medium in combination with the precursor [ 9 , 10 ].…”