16 Feb 2022

Factors Affecting Secondary Metabolite Production In Tissue Culture

Anjali Singh, MS

As a content and community manager, I leverage my expertise in plant biotechnology, passion for tissue culture, and writing skills to create compelling articles, simplifying intricate scientific concepts, and address your inquiries. As a dedicated science communicator, I strive to spark curiosity and foster a love for science in my audience.

Table of Contents

Overview

Plants have significant and prominent roles in both traditional and modern medicines. According to a study, around 80% of the population relies on plant-derived medicines. India is one of the major medicinal plant-producing Asian countries and a reservoir of many high-valued medicinal plants.

Phytochemicals are used in several applications, including cosmetics, pharmaceuticals, and nutraceuticals. You might yourself have observed the increased production of natural or plant-based cosmetics. Nowadays, people prefer to purchase more botanical products compared to chemical ones.

The medicinal activity of plants is imparted by secondary metabolites that are produced in plants for secondary functions, including protection against pathogen attacks and attracting pollinators. These chemical compounds are only produced in plants and therefore the over-exploitation of medicinal plants for several purposes is causing a threat to the extinction of these medicinal plant species.

Tissue culture has been an effective alternative to project these plant species, conserve their genome, and increase their production. The advancement of the technique has also been a fruitful way to increase secondary metabolite production for commercial purposes.

In this article, you’ll learn about factors that affect the production of secondary metabolites in tissue culture.

Factors affecting the production of secondary metabolites in tissue culture

• Media Formulation: Composition and types of media used for the cell culture of plants impact the production of secondary metabolites. For example, the use of MS (Murashige and Skoog) media in the suspension culture of Catharanthus roseus increases the production of serpentine metabolite. The effect of media on metabolite yield varies from one plant species to another.
• Carbon Source: Sucrose is one of the essential components of plant tissue culture media. It has been observed that altering the sugar concentration in the media or using different carbon sources impacts the production of secondary metabolites in some plants.

For example, the use of glucose, instead of sucrose, in the media formulation of Podophyllum hexandrum, increases the production of podophyllotoxin. Whereas, to increase rosmarinic acid, an increase in the sucrose concentration level was found to be an effective approach.

• Nitrogen: Nitrogen is one of the essential components of some tissue culture media, including MS, LS, or B5 media. It has been observed that media containing amino acids and proteins have better secondary metabolite yields. Moreover, the amount of that nitrogen also impacts the production of the metabolites.
• Phosphate: Several studies report that the level of phosphate in tissue culture media affects the production of secondary metabolites. For example, in many plants, it has been observed that increasing the phosphate level in the media increases cell growth, however, reduces the secondary metabolite yields. In Catharanthus roseus, the yield of phenolics and ajmalicine; in Nicotiana tabacum, the yield of caffeoyl putrescine; and in Peganum harmala the yield of Harman alkaloids has been found to enhance after reducing the phosphate level in their tissue culture media.
• Agitation: During cell suspension culture, it has been found that the speed of agitation impacts the cell viability, resulting in impacting the production of secondary metabolites. In Podophyllum hexandrum, a speed of 200 rpm was found to be damaging for plant cells, whereas more viable cells were obtained when speed was in the range of 125- 150 rpm.
• Plant growth hormones: Several studies report that the use of types of hormones and their concentration in the media affect the plant’s growth in tissue culture, in addition to the production of secondary metabolites.

For example, some studies found that 2, 4-D ceased metabolite production, whereas, the addition of NAA and IAA increases the metabolites synthesis. In Catharanthus roseus, auxin alone was able to induce callus production, however, an increase in secondary metabolite production was only observed after growing the plant alone with cytokinin.

• Elicitation: It’s a process of inducing effects like pathogen attacks in plants to induce the expression and functioning of genes that are responsible for the synthesis of secondary metabolites. Both biotic and abiotic elicitors are used for this purpose. It includes jasmonic acid, glucan polymers, glycoproteins, fungal cell materials, UV irradiation, salts of heavy metals, and many other chemicals.

It has been found that in plants Rauwolfia species and Eschscholzia californica, the use of yeast extracts as elicitors increased the production of secondary metabolites. A similar case was observed with using methyl jasmonate with other plants.

• Gas composition: In some cases of secondary metabolites production, it has been observed that the percentage concentration of gasses inside bioreactors impacts the yield. For example, in Panax ginseng, 40% oxygen in the chamber was found to have the highest yield whereas, increasing the concentration to 50% or lowering it to 20-30% resulted in reduced secondary metabolite production.
• Precursor supplementation: Some studies have found that supplementing the cell cultures with compounds that are also produced during secondary metabolite synthesis increases their yield. This process of the addition of such compounds to cell cultures is known as precursor feeding.

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Happy Culturing!!