The effect of hormone concentrations and microsamples on germination, regeneration and suspension culture of fennel

Effect of hormones and explants on calligenesis, regeneration and cell suspension culture in fennel

(Foeniuculum vulgare)

Introduction: Fennel plant (Foeniuculum vulgare) is a herbaceous, stable, aromatic plant from the family of Chetrian, which is used for food and medicine in different regions of Iran. Considering the importance of the compounds of this plant in the pharmaceutical and food industries, the aim of this research was to optimize tissue culture of this plant under hormonal treatments and to increase secondary metabolites. In order to investigate the effect of hormonal treatments on callogenesis and regeneration in fennel plant, three micro-samples (leaf, hypocotyl, root) were selected. The seeds of fennel plant were sterilized in the first step. In the next step, they were transferred to the basic MS culture medium under completely sterile conditions. The desired seeds were placed in the growth chamber at a temperature of 23 degrees Celsius with 16 hours of light and 8 hours of darkness, and after 21 days, when the plant reached a height of about 10 cm, the micro-samples were transferred to the culture medium containing 0.5 and 1 mg of BAP hormone per liter. Callus induction was done by culturing explants on MS medium with 16 hormone treatments including 2,4-D hormone along with BAP and NAA to Hamra Kin. Callus size, callus weight and percentage of callus formation were measured in each of the used treatments. Considering the hyperhormonal traits of 2 and 4 mg/l 2,4-D along with 0.5 mg/l BAP, the best hormonal combinations and the hypocotyl and root of the best explants were identified. In the callus regeneration phase, ½ MS medium without hormones showed the best regeneration medium. In order to study extract production in the cell suspension culture phase, the best treatment was selected in the calligenesis phase and the callus was transferred to the liquid MS cell suspension medium containing 2,4-D (2 and 4 mg/L) along with BAP (0.5 mg/L) and placed on a shaker at 24°C in the dark at 120 rpm. After 2 weeks, mass spectrometry was performed and the production of substances such as alpha-pinene and limonene was reported in the tissue culture medium.

Introduction

Flowers and plants are the most silent creatures and at the same time the most expressive manifestation of the power and greatness of creation. Every leaf of these beautiful creatures is a big book describing monotheism. Flowers and plants not only adorn nature's table with their original and unique shapes and forms, but also make it so full of life force that no other plant can compete with it (Omidbeigi, 1377). However, nowadays the treatment of diseases is mostly done through the use of drugs that have industrial origin and are specially prepared in laboratories, but the use of some of them causes harm to the body and many of them have proven side effects. In the early part of the century, the development of organic synthesis led to the development of the pharmaceutical industry and the treatment of many incurable and often fatal diseases. According to the World Health Organization, more than 80% of people in the world still use herbal medicines The world has a plant origin that is either directly extracted from plants or synthesized based on plant composition. The term medicinal plants does not refer only to pain relievers, but these plants are used as flavorings, beverages, sweeteners, natural colors in the subgroup of food, and are also used as the raw material of cosmetics and health products (Omidbigi, 1376).

In fact, plants that have at least the following characteristics are called medicinal plants:

1- In the body of these plants, special substances They are made and stored as effective substances or secondary metabolites that are used to treat some diseases.  The mentioned substances are made during special and complex biochemical processes and in a very small amount (usually less than one percent of the dry weight of the plant).

2- Often, special organs such as roots, leaves, stems, flowers, fruits, etc. may have the most effective substances, so it is not always possible to consider the whole plant organ as a source of special medicinal substances.

3- Plant organs are harvested, prepared and processed, that is, they are affected by special operations such as separation, crushing, drying, fermentation, etc., and then they are used. Normally, these organs are offered in a traditional way and only by drying them as a perfumery product. The global economy has become stronger. Its increasing consumption is not only limited to developing countries, but it is also considered one of the important health factors of developed countries. In the world, 130 million tons of medicinal plants are bought and sold annually, and about one million pharmaceutical factories have increased in the last few years. According to estimates made in recent years, the value of the global market of herbal medicines, which includes medicinal plants and their products, has grown significantly. Most of the market of medicinal plants in the world is related to the production and supply of secondary metabolites derived from these plants. Secondary metabolites usually have a very high added value, so that the sales value of some of these compounds, such as shikonin [1], digitoxin [2] and perfumes such as jasmine oil [3], varies from several dollars to several thousand dollars per kilogram. Also, the price of each gram of herbal anticancer drugs such as vinblastine [4], vincristine [5], ajmalicin [6] and taxol [7] reaches several thousand dollars. Taxol is one of the medicinal compounds obtained from the bark of the yew tree and is used in the treatment of breast and ovarian cancers. The introduction of medicine allocates a lot of budget to itself, while Iran, having diverse ecological and climatic characteristics, can be one of the rare drawers in the production of various plants, so that Iran has allocated 11 of the 13 climates in the world. And approximately 8000 species of plants, which is equivalent to 2 times the entire flora of Europe, is another characteristic of our country, that this gene bank is of great economic importance. For example, the Philippines earns a huge amount of its income from the sale of medicinal plants and wild plants to other countries (Mirjalili, 2012). Therefore, although we are at the beginning of the development of the medicinal plant industry, we can take a significant part of the world markets with proper planning. At the beginning, we may not be able to compete with countries that have mass-produced their products, but in terms of species diversity and natural production of medicinal species, we do not have many competitors in the world, and in some cases we are even unrivaled. All this depends on knowing the assets of the country and being able to use them optimally. In this case, even up to 5% of the national gross product can be provided in this way (Amiri, 2006).

1-3- Necessity of paying attention to the cultivation of medicinal plants

With the advent of chemical and biological drugs, the role and importance of medicinal plants in providing human health was forgotten. However, with the passage of time, the acceptance of medicinal plants has faced a significant growth. It seems that the people of the world are tired of a series of inadequacies of modern medicine and are increasingly turning to herbal medicines. For this reason, today about 50% of the medicines produced in the world are of natural origin, which are used as medicine with changes, half of this amount is obtained from mineral, animal and bacterial sources, and the other half is of plant origin. For example, all hormones used are plant based and are obtained from different plants such as Mexican potato, fenugreek, etc. Likewise, compounds such as vinblastine and vincristine, which are anticancer drugs, are obtained from plants, or cardiac glycosides are considered among this group of drugs (Qasemi Dehkordi and Taleb, 2010). Medicinal plants have better compatibility with the body due to their natural nature and the presence of homologous medicinal compounds together, and they usually have no unwanted side effects, so they are especially suitable for long-term use and chronic diseases.