Factors Affecting Cell Biomass and Flavonoid Production of Ficus deltoidea var. kunstleri in Cell Suspension Culture System

Zainol Haida,Ahmad Syahida,Mahmood Maziah,Syed Mohd Ariff,Mansor Hakiman

doi:10.1038/s41598-019-46042-w

Abstract

A study was conducted to establish in vitro culture conditions for maximum production of biomass and flavonoid content for Ficus deltoidea var. kunstleri, locally named as Mas Cotek, known to have a wide variety of potential beneficial attributes for human health. Size of initial inoculum, cell aggregate and initial pH value have been suggested to influent content of biomass and flavonoid for cell suspension culture in several plant species. In the present study, leaf explants were cultured by cell suspension culture procedures in MSB5 basal medium supplemented with predetermined supplements of 30 g/L sucrose, 2.75 g/L gelrite, 2 mg/L picloram and 1 mg/L kinetin with continuous agitation of 120 rpm in a standard laboratory environment. Establishment of cell suspension culture was accomplished by culturing resulting callus in different initial fresh weight of cells (0.10, 0.25, 0.50, 1.0, and 2.0 g/25 mL of media) using similar basal medium. The results showed that the highest production of biomass (0.65 g/25 mL of media) was recorded from an initial inoculum size of 2.0 g/25 mL media, whereas the highest flavonoid (3.3 mg RE/g DW) was found in 0.5 g/25 mL of media. Cell suspension fractions classified according to their sizes (500–750 µm, 250–500 µm, and <250 µm). Large cell aggregate size (500–750 µm) cultured at pH 5.75 produced the highest cell biomass (0.28 g/25 mL media) and flavonoid content (3.3 mg RE/g DW). The study had established the optimum conditions for the production of total antioxidant and flavonoid content using DPPH and FRAP assays in cell suspension culture of F. deltoidea var. kunstleri.

Highlights

Ficus deltoidea Jack, locally known as Mas Cotek, is an evergreen shrub traditionally used in Malaysia as a remedy to treat cardiovascular diseases and diabetes[1]
High demand for these plant-based products has led to over-harvesting of wild species and the eventual decrease of many medicinal plant species in nature leading to a definite limitation to their availability[7]
This was followed by the initial inoculum size of 0.25, 0.5 and 1.0 g/25 mL of media which produced 0.12, 0.28 and 0.56 g/25 mL of dry weight respectively

Summary

Introduction

Ficus deltoidea Jack, locally known as Mas Cotek, is an evergreen shrub traditionally used in Malaysia as a remedy to treat cardiovascular diseases and diabetes[1]. It has been reported that polyphenols, flavonoids and tannins were present in F. deltoidea extract but alkaloids, triterpenes and saponins were absent[5]. Phytochemical compounds such as polyphenols, flavonoids and tannins are the natural source of antioxidant. Plant cell suspension culture is one recent technique of producing wide variety of secondary metabolites in large quantities in shorter period of time. In such culture system, nutrient components in basal media play important roles in the initiation and production of secondary metabolites by controlling enzyme activities as well as gene expression[9]. The present study was undertaken to investigate the effect of initial size of inoculums, cell aggregate and pH of medium in a cell suspension culture of Ficus deltoidea var. kunstleri with an aim of establishing the level for optimum production of biomass and flavonoid

Methods

Results

Conclusion