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In summer 2020, SIA “Field and Forest” in cooperation with researchers from the Institute for Environmental Solutions (IES) launched the study on commercial propagation of endangered medicinal and aromatic plants by using tissue culture method. This approach will provide an opportunity to cultivate endangered species in organic farming.


Along with the increased pharmaceutical, cosmetology, and food industry demand for medicinal and aromatic plants (MAPs), populations of many wild species are decreasing. Therefore, researchers are eager to study the cultivation of endangered plant species in organic farming. This solution will reduce pressure on wild populations, as well as ensure a sufficient amount of high-quality plant-based materials for different industries.

Three endangered wild medicinal plant species with high market potential were selected for this research – Siberian ginseng (Eleutherococcus senticosus Maxim.), common yew (Taxus baccata L.) and sea holly (Eryngium maritimum L.). During the 3-year period, IES researchers will develop efficient and economically viable propagation methods of these plants for commercial use in organic farming.

In the following interview, IES leading researcher Dr Ieva Mežaka highlighted the progress and the importance of this research.



IES: Why is it necessary to do this research?

Dr Ieva Mežaka (I.M.): 90% of medicinal and aromatic plants are harvested in wild and only 10% are cultivated. Increasing harvesting of MAPs in the wild, along with the loss of natural habitats, bring a heavy burden on wild populations, therefore, many of them are endangered. Meanwhile, the human population is growing and so is the market demand for MAPs. In this research, we are looking for ways to grow medical and aromatic plants more effectively – by cultivating them in organic farming. Another important challenge is to seek for the growing technologies that could provide the highest harvests and concentration of active compounds in plants.



 

IES: Why did you choose these 3 MAPs – Siberian ginseng (Eleutherococcus senticosus Maxim.), common yew (Taxus baccata L.) and sea holly (Eryngium maritimum L.)?

I.M.: The main reason was the high market demand for these MAPs due to which their wild populations are endangered. During this research, we will develop technologies for cultivation of these endangered MAPs in organic farming, thus decreasing the pressure on wild populations. Additional factors for selecting these plants were their unifying vulnerabilities – prolonged seed germination, low seed production and slow rooting of cuttings.

IES: You mentioned that the selected MAPs have a high market demand. Where exactly one can use active compounds of these plants?

I.M.: 60% of all produced pharmaceuticals contains active compounds extracted from medicinal and aromatic plants. Wild Siberian ginseng, common yew and sea holly have a high concentration of different active compounds widely used in pharmaceutical, food and cosmetics industries. For example, common yew is used for the development of anti-cancer drugs. Moreover, research shows that Siberian ginseng has a stimulating and anti-depressive effect, but Sea holly is used for liver and kidney disease cures.

IES: Please explain this plant tissue culture propagation method?

I.M.: It is a biotechnology method that allows us to propagate plants by using tissue cultures. We grow these plants in laboratory test tubes, but we do not change their genetics. This method can be divided into four steps:

First step. We start with seed germination then we grow a plantlet.

Second step. Then we do micro-propagation. We take that plantlet that we grew and cut it in pieces. From these pieces, we can re-grow new plantlets and continue this process in geometrical progression.

Third step. Rooting of the plantlets propagated in the second step.

All this process is done in controlled and sterile conditions. Micro-propagation and rooting require adaptation of each medium composition that we are providing to the plantlets. This includes the composition of micro-and macro-elements, vitamins, growth regulators, various other additives, and their concentrations. This is an important process because it determines the growing conditions of the plant and plantlet development.

Fourth step. Adaptation of plants grown in laboratory conditions to conventional growing conditions in the soil. Plants that are propagated in these circumstances where we provide a suitable environment – nutrients, humidity, suitable temperature etc., are relatively fragile to external impact factors.

 

IES: Why did you choose this method instead of other easier approaches?

I.M.: All three of the researched plants under normal propagation conditions germinate slowly and unevenly. It takes up to 18 months to sprout Siberian ginseng seeds and they need stratification – a simulated set of environmental effects in controlled conditions which accelerate seed sprouting. For example, by subjecting seeds to cold treatment we simulate the period that seed experiences under natural conditions during winter. Plant tissue culture method in the propagation process allows us to exclude the long seed sprouting procedure. We sprout the gathered seeds once at the beginning of the research and afterwards, we work only with plantlets and micro-propagation. This process is much faster than other more traditional propagation methods, therefore more suitable for commercial cultivation. Additionally, this method allows us to do a chemical analysis of plantlets, thus allowing us to understand which plants have a higher concentration of active compounds. Then we can choose which ones to propagate for commercial use.


 

IES: Is it planned to propagate populations harvested in Latvia?

I.M.: In Latvia wild populations of common yew and sea holly can be found but wild Siberian ginseng is common in Russia and China. For the last few decades, private farmers have proven that it is possible to cultivate Siberian ginseng in natural conditions of Latvia. Within this research, we were interested to gather wild population samples as well as ones that are grown by farmers from Latvia and other natural distribution areas of these plants.

We launched this research in summer of 2020. So far, we have already implemented sampling campaigns where we collected different parts of plants (leaves, branches, roots) and seeds. In Latvia, these plants are endangered, therefore we needed to get special permission approved by the Nature Conservation Agency. We have gathered samples of all three species found in Latvia (wild species and samples from private growers). We also carried sampling expeditions to Saaremaa and Kihnu islands in Estonia. Also, for these expeditions, we needed to get special permission from the Environmental Board.

We were planning to carry sampling campaigns in other natural distribution areas of these plants, but our plans were disrupted by COVID-19 pandemic. We adapted to this situation and found other solutions for the collection of samples. In seed exchange programs we were able to gather several samples from the collections of national botanical gardens, universities and private growers from Latvia and other countries.

 

IES: What have you planned for the next research periods?

I.M.: Sample gathering campaigns are concluded. We have started the work with seed stratification and introduction of seeds to plant tissue culture. Researchers in the laboratory began the assessment to find the best approach of chemical analysis for all three researched MAPs. We are developing a methodology and preparing extracts for chemical analysis. That will help us understand what kind of active substances are in the researched plants and how they differ between populations. Thus, we hope to find out which of the researched populations are the most valuable ones.

Research on plant tissue culture application for commercial propagation of endangered medicinal plants is developed as a part of the European Regional Development Fund programme 1.1.1 “Improve research and innovation capacity and the ability of Latvian research institutions to attract external funding, by investing in human capital and infrastructure” 1.1.1.1. measure “Support for applied research”, Nr. 1.1.1.1/19/A/083.

More about the project here.

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In summer 2020, SIA “Field and Forest” in cooperation with researchers from the Institute for Environmental Solutions (IES) and SIA “Alternative Plants” launched the study to develop high added value bioactive cosmetic ingredients from by-products of medicinal plant processing and plant cell cultivation.

The growing human population also creates an increasing demand for natural resources for food, medicine, pharmacy, and other industries. The use of underutilised industrial by-products could help Europe’s leading industries to develop high added-value products, meet the growing demand and move towards more sustainable processes.

For this research by-products of medicinal Chamomile (Matricaria) processing and in-vitro cell culturing of Dragonhead (Dracocephalum), European gooseberry (Ribes) and Juniper (Juniperus) were chosen because researchers saw a potential in them to develop high added-value products.

In the following interview, IES leading researcher Dr Ilva Nakurte highlighted the progress and importance of this research.



IES: Why is it necessary to do this research?

Dr Ilva Nakurte (I.N.): Most by-products coming from agriculture, cosmetics, food and drink, as well as chemical industries are not utilized and end up in municipal landfills, thus causing serious environmental, economic, and social problems. Currently, by-products are handled as waste, while they could be used as resources to create new revenue streams and open doors for new niche markets. Spinning the mindset and business approaches towards using more innovative, science-based solutions for by-product utilization will have a positive impact on the transition towards a sustainable economy.

Medicinal and aromatic plants have active compounds that are highly demanded by pharmaceutical, cosmetics, and food industries. While active compounds can be artificially synthesised, considering increasing demand, the focus should be on compounds extracted from natural resources. Therefore, during this research, we will obtain active compounds from different industry by-products and waste to develop bioactive cosmetic ingredients.

 

IES: What do you consider as the main valuable resource that you will obtain from by-products?

I.N.: When people hear the word waste, the first association in their mind is household waste. In this research we will work with two groups of organic medicinal plant processing by-products:

– Medicinal and aromatic plant processing by-products from medicinal chamomile essential oil extraction provided by organic farming experts of SIA “Field and Forest”.


– Plant tissue cultivation by-products of cosmetic production provided by SIA “Alternative plants” company.

 

Before the start of this research, we already knew that these medicinal and aromatic plant processing by-products contain high concentration levels of valuable compounds. Within this research, we will develop technologies for bioactive compound extraction (with suitable biorefinery methods) from above-mentioned by-products. After extraction active compounds will be categorized and further used in the development of different valuable product prototypes for the cosmetic industry.

IES: Please explain what is biorefinery and how it helps to extract active compounds from medicinal and aromatic plant processing by-products?

I.N.: A biorefinery is defined as the optimised processing of biomass for extraction of new raw materials.

We are convinced that in our research selected by-products materials contain bioactive compounds. Therefore, the main task of the biorefining processes used in our project is to understand what and how much active substances they contain, and how to extract them in the most effective ways. The processes of the biorefinery method that we will implement in this study can be divided into three main steps:

First step. We are going to search for the most suitable method for the extraction of bioactive compounds. We have chosen the most environmentally friendly and, at the same time, efficient active compound extraction method – the supercritical fluid extraction method.

Second step. Reuse of waste is an environmentally friendly action. In this research we are trying to find the most environmentally friendly extraction reagents. We found it inappropriate to use chemical reagents for extraction that are harmful to the environment and human health. For the supercritical fluid extraction method use only carbon dioxide (CO2), in combination with ethanol and/or water. To extract the highest concentration of active compounds from each group of by-products, it is important to find the most suitable combination of these three reagents.

Third step. Fractionation of polar and non-polar compounds. All active compounds that we extracted from the by-products need to be divided into fractions. As a result of extraction, we will obtain the whole set of biologically active substances, and there is no unique method that would be able to separate them from each other in one go. The supercritical fluid extraction method allows us to systematically divide this group into fractions, such as carbohydrates, proteins, antioxidants, fatty acids, volatile substances. Fractionation allows us to understand which groups of bioactive compounds we were able to extract and how high are the concentrations of these groups. If we understand the value of each of the active compound groups, we can assess whether the size of the class is large enough to make its extraction economically viable.

IES: What are the next steps after the extraction is done and active substances are divided into fractions?

I.N.: Once the extracts have been divided into fractions, the next step is the chemical characterization of extracts and fractions. The chemical characterization will be done in IES laboratory using state-of-the-art analytical equipment and extracts will be classified in such groups as sugars, amino acids, glycosides, tannins, phenolic compounds, flavonoids, etc.

An assessment of the microelement composition of the obtained fractions and extracts will also be performed, as we are interested not only in the bioactive substances contained in these medicinal plant by-products but also in the microelements, which is a very hot topic nowadays.

IES: What will you do with the biologically active compounds that you will acquire?

I.N.: Next step will be the assessment of the acquired bioactive compounds. During discussions with all research partners, we will decide which compounds and from which by-products have the highest potential in cosmetic product development.

Our research partners SIA “Alternative Plants” have extensive and long-term experience in cooperation with various cosmetic manufacturers, therefore they will start the combination of biorefinery fractions and extracts to generate a composite of high-value cosmetic ingredient. This ingredient will be tested on human skin cells to assess the impact. If human cell tests will show positive results, we will continue the development of a product prototype for the cosmetics industry.

IES: Is it planned to produce an actual product within this research?

I.N.: No, we have not planned to develop a product for the cosmetics industry during this research. The outcome will be intellectual property that will belong to all three project partners. Our research activities will certainly not stop. Research on natural resource-based by-products for the development of new product prototypes can be interesting for the organic cosmetics industry, as well as many others.

IES: Is this extraction of active compounds from plant processing by-products an innovative approach?

I.N.: Of course, we are not the first ones that have thought about the extraction of bioactive compounds from plant processing by-products that, otherwise, would go to waste. However, In Latvia, we are one of the pioneers that focus on large-scale medicinal plant by-product repurposing.

In scientific literature, we can find similar examples of organic plant by-product extracting even with the same supercritical fluid extraction method. But researchers usually do not share specific techniques that are vital in the research process.

Within this study, we will evaluate the extraction methods and adapt them to each group of by-products in order to find the most suitable and effective solutions.

 

IES: This research was launched in July 2020. Which activities have you already started?

I.N.: We have started to test the first water and ethanol extracts from six SIA “Alternative Plants” different lyophilized plant cell culture by-products and from SIA “Field and Forest” chamomile processing by-products – white petals, pollen (pulp) and distillation waste. We have started testing the reagents of the supercritical fluid extraction method. For each group of by-products considered in the study, it is necessary to find the right combination of carbon dioxide, water and ethanol, as well as the most suitable extraction temperature and time. We try to find out these factors in a series of experiments, to get the most effective extraction results.

Experimental extraction of non-polar fractions from distillation by-products with non-polar liquefied gas – florasol extraction – was also performed. For both non-polar and polar fractions, a qualitative assessment of the effect of distillation time on the separation of fractions from distillation by-products has been performed. We can already conclude that the hypothesis put forward in our project has proved to be true, and high concentrations of various bioactive substances can be found in the production by-products tested so far.

IES: What have you planned for the upcoming research periods?

I.N.: We will continue to adapt the extraction method for each group of by-products. We will continue the fractionation of the obtained polar and non-polar compounds and the characterization of these fractions. That will allow us to find out which fraction contains the most valuable bioactive compounds and which of them would be worth researching further.

Research “A biorefinery approach for the development of bioactive cosmetic ingredients from by-products of medicinal plant processing and plant cell cultivation” (1.1.1.1/19/A/075) is developed as a part of the European Regional Development Fund programme 1.1.1.1 measure “Support for applied research” and specific objective 1.1.1 “Improve research and innovation capacity and the ability of Latvian research institutions to attract external funding, by investing in human capital and infrastructure”

More about the project here.

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