Evaluation of Manganese Chloride's Effect on Biosynthetic Properties of In Vitro Cultures of Eschscholzia californica Cham.

Andrea Balažová,František Bilka,Pavel Mučaji,Ivana Holková,Júlia Urdová,Vladimír Forman,Branislav Horváth

doi:10.3390/molecules23040971

Abstract

The basal production of secondary metabolites in medicinal plants is limited. One of the effective approaches that encourages plants to produce a remarkable amount of precious compounds is an application of elicitors. Our work was focused on the elicitation of Eschscholzia californica Cham. suspension cultures using various concentrations of MnCl2 (5; 10; 15 mg/L) with the aim of evaluating its effect on sanguinarine, chelerythrine, and macarpine production and gene expression of enzymes involved in the biosynthesis of mentioned secondary metabolites (BBE, 4′-OMT, CYP80B1) or in defense processes (LOX). Suspension cultures were exposed to elicitor for 24, 48, and 72 h. The content of alkaloids in phytomass was determined on the basis of their fluorescence properties. The relative mRNA expression of selected genes was analyzed using the ΔΔCt value method. PCR products were evaluated by melting curve analysis to confirm the specific amplification. Our results demonstrated that Eschscholzia californica Cham. cell suspension cultures evince sensitivity to the presence of MnCl2 in growth media resulting in the increased production of benzophenanthridine alkaloids and gene expression of selected enzymes. Manganese chloride seems to be a potential elicitor supporting natural biosynthetic properties in plant cell cultures and can be applied for the sustained production of valuable secondary metabolites.

Highlights

The role of secondary metabolites in plants was not clear for a long time
Our results demonstrated that Eschscholzia californica Cham. cell suspension cultures evince sensitivity to the presence of MnCl2 in growth media resulting in the increased production of benzophenanthridine alkaloids and gene expression of selected enzymes
California poppy suspension cultures were prepared from friable callus proliferated on a paper bridge in Murashige–Skoog (MS) medium supplemented by kinetin (2 mg/L) and 1-naphtylacetic acid (NAA) (1 mg/L)

Summary

Introduction

The role of secondary metabolites in plants was not clear for a long time. Secondary metabolites are generally not essential for the growth, development, or reproduction of plants but play a major role in the adaptation of plants to their surrounding environment and in overcoming stress [1,2,3].The involvement of plant secondary metabolites in defense processes is not their single benefit; many of them possess immense biological activities. The role of secondary metabolites in plants was not clear for a long time. The effects of medicinal plants on human or animal organisms have been discovered in ancient times based on empirical experience [4] before the isolation and characterization of the active compounds responsible for their therapeutic effect [5,6]. Current drug development is based on the synthetic preparation of active compounds, Molecules 2018, 23, 971 ; doi:10.3390/molecules23040971 www.mdpi.com/journal/molecules. Molecules 2018, 23, 971 the importance of natural products in pharmacy cannot be fully replaced by synthetic drugs. Isolation of desired natural products from wild-growing or cultivated plants is often the only way to obtain appreciable amounts because their chemical synthesis is either extremely difficult or non-profitable [7]

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Conclusion