Eurasian Journal of Applied Biotechnology

OBTAINING ASEPTIC CULTURE IN VITRO OF RHODIOLA SEMENOWII L. FROM VARIOUS TYPES OF EXPLANTS

O. Raizer — 2024-11-12

In vitro cultivation of callus cells allows for the stable production of secondary metabolites, being an alternative to the use of medicinal plant biomass from natural populations. Standardization of cultivation protocols and technological processes for the synthesis of secondary metabolites in vitro conditions determines significant interest in the use of tissue culture methods (PTC – Plant Tissue Culture). The aim of the study was to obtain an aseptic culture of Rhodiola semenowii in vitro from various types of explants, as well as the effect of the hormonal composition of the nutrient medium on the induction of callusogenesis in the culture of isolated organs of R. semenowii. As a result of the research, the optimal mode of obtaining R. semenovii aseptic cultures from various explants was determined. It has been shown that the use of 70% EtOH in combination with 0.01% HgCl₂ or 20% NaClO makes it possible to obtain aseptic cultures from any type of R. semenovii explants. It has been established that the type of explant is R. semenovii influenced both the frequency of callus formation and the beginning of its formation. The least optimal types of R. semenovii explants are stem and rhizome segments, the frequency of callusogenesis varied from 5.8% to 75.6%. The induction of callusogenesis also depended on the hormonal composition of the nutrient medium: the highest percentage of callus formation in all types of explants was on media containing 0.5mg L⁻¹ thidiazuron (TDZ) + 1.0 mg L⁻¹1-naphthaleneacetic acid (NAA) and 0.5mg L⁻¹2,4D + 2.0 mg L⁻¹ 6-Benzylaminopurine (BAP).

INFLUENCE OF ALTERNARIA SPP. FUNGI METABOLITES ON DEVELOPMENT OF WHEAT SPROUTS

I.V. Rukavitsina — 2024-11-15

The study purpose was in vitro investigation of phytotoxic activity of culture broth filtrates (CBFs) of five Alternaria Spp. fungi isolates on development of wheat sprouts. Fungi isolates were received from wheat seeds cultivated under conditions of Northern Kazakhstan. According to the study findings, culture filtrates of all the isolates studied varied with regard to their influence on wheat sprout development. Stimulant effects on seed germinating power were revealed with 14- and 21-day CBFs of A. infectoria (7/4), 14-day CBF of A. brassicae (7/2) and 21-day CBF of A. porri (8/1). It was found that long-term cultivation of fungi on a culture medium contributed to media pH modification towards alkalization due to production of metabolites of various chemical nature, including toxins, which could trigger growth inhibition of radicles and sprouts. However, the CBFs were predominantly low-toxic or non-toxic. Under effect of the 14-day CBF, significant positive correlation was identified between pH values and sprout lengths (0,86±0,25), while negative correlation was found between pH values and germinating power (-0,75±0,33). Change of pH value on the 21^st day significantly affected radicle length (-0,92±0,20). Based on the findings it can be concluded that the 14-day CBF of all the Alternaria fungi isolates studied was most toxic for wheat sprouts.

IDENTIFICATION OF SECONDARY METABOLITES BY GAS CHROMATOGRAPHY WITH MASS SPECTROSCOPY IN CALLUS TISSUES OF CISTANCHE DESERTICOLA Y.C. MA

M.Y. Sutula — 2024-11-18

The aim of this study is to analyze the types of secondary metabolite compounds found in cistanche (Cistanche deserticola) plants. Methanolic extracts of callus tissues and shoots (stolon) of cistanche were obtained by lyophilic drying and grinding to powdered state using 80% methanol. Secondary metabolites within cystanche callus were first identified by GC-MS (gas chromatography-mass spectroscopy). The identification results of the methanolic extracts revealed various types of compounds including fatty acids (hexadecanoic acid, methyl ester); phenolic compounds and their derivatives (phenol, 2,4-bis(1,1-dimethylethyl)-, 2-methoxy-4-vinylphenol, homovanillyl alcohol, phenol, 2,6-dimethoxy;) sterols (γ-sitosterol); sugars, glycosides and other carbohydrates (ethyl α-D-glucopyranoside, sucrose), glycols and their derivatives (tetraethylene glycol, diethylene glycol monodododecyl ether), etc. Some of the detected compounds may be precursors or by-products of phenylethanoid glycosides biosynthesis. We also found a correlation between callus color and its chemical composition of secondary metabolites. The data can be used for further optimization of C. deserticola cultivation methods to increase the content of target secondary metabolites.

POTENTIAL FOR ACCELERATED REPLICATION OF FRAXINUS SOGDIANA USING IN VITRO CULTURE AND IDENTIFICATION OF A RELIC TREE USING DNA-BARCODING

M.K. Amangeldinova — 2024-11-19

Four species of Fraxinus including F. angustifolia subsp. syriaca (Boiss.) Yalt., F. sogdiana Bunge, F. pennsylvanica Marshall and F. americana L. grow in Kazakhstan. The study aimed to determine the conditions for awakening endogenous dormancy in vitro of F. sogdiana. In the control group without vernalization, the sterile culture yield at room temperature was 30%. However, with cold treatment, this yield increased to 60%, which is twice as high. It was observed that on MS medium without the addition of hormones, 22 microshoots were obtained from unwounded seeds, whereas 39 microshoots were obtained from wounded seeds during microclonal propagation.Phylogenetic relationships and species identification were analyzed using three DNA barcoding markers, comparing F. sogdiana with species from the NCBI database. rbcL and ITS markers did not show satisfactory interspecific genetic variability among F. species. Notably, matK gene sequences effectively differentiated F. sogdiana from other species. These results provide fundamental information that is valuable for future biotechnological and molecular studies.

ASSESSMENT OF BIOLOGICAL RISKS IN INFECTIOUS DISEASES TO ENSURE BIOLOGICAL SAFETY

B.Sh. Myrzakhmetova — 2024-11-19

The territory of the Republic of Kazakhstan is unfavorable for a number of infectious diseases of animals and humans that pose an enzootic biological threat. Such nosological units include anthrax, blackleg, rabies, leukemia, brucellosis, tuberculosis, pasteurellosis, sheep pox, camel pox, CCHF, highly pathogenic avian influenza, foot-and-mouth disease, tick-borne encephalitis, influenza A and B, infectious hepatitis, Newcastle disease, seasonal flu, coronavirus infection COVID-19, etc., some of which affect only animals or humans, and some affect both animals and humans. While some of these diseases are natural focal and pose a constant biological threat, others appear from time to time, penetrating from neighboring countries or develop as a result of movement from the wild - reservoirs and hidden carriers of pathogens. A number of exotic diseases pose a real threat of penetration from outside the country due to interstate movements of people, animals, feed and animal products.