Effect of methyl jasmonate and salycilic acid synergism on enhancement of bilobalide and ginkgolide production by immobilized cell cultures of Ginkgo biloba

Agus Sukito,Sanro Tachibana

doi:10.1186/s40643-016-0101-0

Abstract

Bilobalide and ginkgolides are reported to be present only in Ginkgo biloba. However, only trace amounts of bilobalide and ginkgolides are contained in the ginkgo leaves. Nowadays, there has been considerable interest in plant cell cultures as a potential alternative to traditional agriculture for the industrial production of secondary metabolites. Much effort has been put into the use of in vitro cultures as one attractive biotechnological strategy for producing bilobalide and ginkgolides of commercial interest. The aim of this study was to enhance the production of bilobalide and ginkgolides A, B, and C in cell cultures of G. biloba using immobilized cell cultures and the process of elicitation. Based on the effect of the immobilization components on the free cell suspension cultures, it was considered that the jute fiber acted as an elicitor and forced the cells to release the product into the culture media. The resulting biomass was approximately 1.4 times higher than in the cell suspension cultures, and the production of bilobalide and ginkgolides A, B, and C increased 5.0, 3.3, 6.1, and 4.1 times, respectively. Eliciting with methyl jasmonate (MJ) and salicylic acid (SA) in the immobilized cells enhanced bilobalide and ginkgolides A, B, and C, compared with the unelicited controls. The highest accumulation was observed using a combination of 0.1 mM MJ + 0.1 mM SA in the immobilized cells, which produced 1.78, 1.95, 2.05, and 2.95 times more bilobalide and ginkgolides A, B, and C, respectively, than the controls. The positive effects of immobilized cell cultures using jute fiber and the synergism effect of SA and MJ on immobilized cells of G. biloba appear to be the optimal conditions for continuous in vitro production for commercial purposes. This is the first report on analyzing the effects of jute fiber as immobilized cell material on G. biloba cell cultures and the synergism of MJ and SA on bilobalide and ginkgolide production.

Highlights

Bilobalide and ginkgolides are reported to be present only in Ginkgo biloba
The problem of low secondary metabolite production caused by lack of cell-to-cell contact in plant cell cultures is still found with those techniques
Plant material and the explants Ginkgo biloba leaves were collected from the garden of the Faculty of Agriculture, Ehime University, Japan, Determination of dry weight Dry weight was determined by filtering the biomass through normal filter paper and the cell clumps were removed from the immobilization materials to obtain the fresh weights of biomass for each flask

Summary

Introduction

Bilobalide and ginkgolides are reported to be present only in Ginkgo biloba. only trace amounts of bilobalide and ginkgolides are contained in the ginkgo leaves. The aim of this study was to enhance the production of bilobalide and ginkgolides A, B, and C in cell cultures of G. biloba using immobilized cell cultures and the process of elicitation. Immobilization of plant cells is a method used in plant cell cultures to induce secondary metabolite production. Several studies have reported that immobilized plant cell cultures show an increase in secondary metabolite production compared with cell suspension cultures (Gilleta et al 2000; Premjet and Tachibana 2004). Since immobilization of suspended cells potentially causes toxicity of polyurethane foam (Vorlop et al 1992) and diffusional resistance of gel matrices (Ganguly et al 2007), cells immobilized with loofa, sisal, and jute provided higher amounts of biomass with enhanced alizarin and purpurin content in immobilized Rubia tinctorum L. in a suspension culture (Nartop et al 2013)

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