Pentacyclic triterpenoids and polyphenols accumulation in cell suspension culture of Chaenomeles japonica (Thunb.) Lindl. ex Spach

MAŁGORZATA KIKOWSKA, AGATA WŁODARCZYK, ANNA STOCHMAL, JERZY ŻUCHOWSKI, BARBARA THIEM

Abstract

Introduction: Callus and cell suspension cultures are widely applied in investigation of production of highvalue secondary metabolites, which may be used as cosmeceuticals, nutraceuticals and pharmaceuticals. Plant cell cultures are promising alternative to intact plant sources for the production of plant-derived drugs of industrial importance. Objective: The aim of the study was to (i) initiate the cell suspension culture of Chaenomeles japonica from homogenous and uniform callus, (ii) stabilize the selected line and (iii) verify its ability to produce the desired groups of secondary metabolites – pentacyclic triterpenoids and polyphenols. Methods: To establish a cell suspension culture, stabilized and homogeneous callus was selected. Cell cultures were systematically passaged every 2 weeks to fresh liquid medium with the same composition. Biomass from cultures at the growth phase and stationary phase was designated for phytochemical research. UHPLC-DAD-MS analyzes were performed. At the same time, their macroscopic and microscopic observations were carried out. Results: Cells of suspension culture line A2 were characterized by the intense divisions. Cell culture extracts (both from the growth phase and stationary phase) contained pentacyclic triterpenoids. In addition, phenolic compounds (chlorogenic acid and proanthocyanidins type B) and in a small amount also epicatechin are present in the extract of the cells harvested from the growth phase. In the present studies, three pentacyclic triterpenoids were detected and quantified in the extracts of cell suspensions and callus line A2. Ursolic and oleanolic acids were the main triterpenoids in the studied extracts. The cell suspension culture from the growth phase exhibited the highest content of ursolic, oleanolic, and betulinic acid (separately and together). Conclusion: The cell suspension culture of Chaenomeles japonica is a promising source of pentacyclic triterpenoids.
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