57
RESEARCH ON METHODS FOR INTRODUCING WHITE SEED (
SILYBUM
MARIANUM
) IN VITRO CULTURE
Sattarov A. N.
Kayumov F.S.
Tashkent Pharmaceutical Institute, Tashkent, Republic of Uzbekistan
https://doi.org/10.5281/zenodo.15210804
Abstract:
After gaining independence, the Republic of Uzbekistan, along with reforming
all sectors, paid great attention to the development of the pharmaceutical industry. Today, the
pharmaceutical industry is one of the fastest growing sectors in our country. It is aimed at
finding new bioactive substances, creating medicines from them, and applying them in medical
practice to meet the needs of our country's population in medicines.
Research objective:
Introduction of the milk thistle plant,
Silybum marianum
, into in
vitro culture and development of optimal conditions for microclonal propagation of the plant.
Methods and techniques:
Sterile seedlings are used to obtain explants for callus and
tumor cultures, microclonal propagation, and the study of hormonal regulation. Seeds are sown
either in water or in a nutrient medium for germination. Before sterilization, plant objects are
thoroughly washed with running water, sometimes with detergents, and cleaned of excess
tissue. The skin is removed from tubers and roots, and the shoots and buds are removed from
the plants. Plant explants are sterilized with solutions of active chlorine (chloramine,
hypochlorite Na), bromine (bromine water), tween-20, hydrogen peroxide, alcohol, silver
nitrate, diacid, and antibiotics. It is necessary to choose a concentration of sterilizing agents that
will only harm the seeds themselves, do not inhibit their growth, and ensure maximum sterility.
Ethyl alcohol is often used to pre-sterilize the material by wiping the surface or immersing it in
absolute alcohol for a few seconds. Sometimes such sterilization is sufficient, it is used when
working with fruits, seeds, buds, nodes. Calcium hypochloride (bleach) is used in the form of a
5-7% solution to sterilize buds, buds, flowers, seeds, buds for 5-8 minutes. Sodium hypochlorite
is used as a 0.5-5% solution to process any explants for 1-20 minutes. This substance is toxic to
cells, therefore the sterilization time and concentration are selected experimentally.
Results:
White carrion plants rooted in the laboratory were successfully transplanted
into sandy peat substrate (90-98% of plants continued to grow). White carrion plants
transplanted into natural conditions grew rapidly, forming a well-developed root system, many
lateral shoots and healthy leaves within a month.
Conclusions:
The initial experiments on microcopying the white carrak plant consisted
of the following: first, the plant seeds were grown in a nutrient medium, then explants were
taken from the sprouted seedlings and grown in nutrient media of different concentrations.
Then, with the help of phytohormones and vitamins, their leaf, shoot and root growth were
carried out, and the best growth conditions were compared. Finally, the plants that grew well
and had a strong root system were adapted to the external environment. Nodular explants grow
faster than leaf explants because they develop shoots more quickly. When certain hormone
balances are maintained in VarIII medium, the explants develop many shoots and roots
simultaneously. Peroxidase activity and auxin content change during the rooting process.
Polyamines can be used as indicators to help determine the rooting process more quickly.
58
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