scholarly journals Characteristics growth of mycelium of mushroom shiitake Lentinula Edodes (Berk.) Pegler in vitro

2019 ◽  
pp. 20-25
Author(s):  
T. Ivanova ◽  
N. Voloschuk
Keyword(s):  
Growth Rate ◽  
Light Microscopy ◽  
Pure Culture ◽  
Nutrient Medium ◽  
Sodium Selenite ◽  
Lentinula Edodes ◽  
Ph Value ◽  
Nutrient Media ◽  
Microbiological Methods

Goal. The purpose of this work is to study the growth of the mycelium of the fungus Lentinula Edodes (Berk.) Pegler on nutrient media of different composition and also to study the features of the use of Avatar-1 micro fertilizer and sodium selenite. Methods. Biotechnological research methods. Sodium selenite (Na2S eO3) at a concentration of 1.0 mmol / l was also used. Pure culture of the mushrooms had a very dense structure and white color. In our work, we used biotechnological methods — obtain this by subculturing the L. edodes strain in vitro; microbiological methods — obtaining pure culture of the fungus, the study of the cultural properties of the colonies. We set the pH value (pH) of nutrient media at the beginning and at the end of incubation. We applied mycological methods to measure the speed, density of growth and dry mass of mycelium. we used the light microscopy method. We performed statistical data processing.Method of light microscopy. Results. The experiments showed about acceleration of mycelial growth, mass and the greatest yield of mycelium L. edodes were on a nutrient medium with microfertilizer Avatar-1. In the experiment, it was found that the maximum overgrowth of the medium by mycelium occurs at 7 days. We have been proved that in the «Avatar-1» nutrient medium there was an increase and consolidation of bifurcated hyphae and buckles of L. edodes. There was even germination of mycelium that did not have too thin or thick hyphae. The dependence of growth rate on the type of nutrient medium. Also, the of doses of the drug, which effectively influences the technology of obtaining primary mycelium L. edodes. Conclusions. The dependence of the growth rate on the type of nutrient medium, the dose administration rate, as well as the cultivation regimes, that effectively influence and cheapen the technology of obtaining the primary mycelium L. edodes is demonstrated.

scholarly journals Designing a Nutrient Medium for the Accumulation of Microbial Mass of Listeria

2020 ◽  
Vol 5 (4) ◽  
pp. 60-66
Author(s):  
N. M. Khaptanova ◽  
S. V. Lukyanova ◽  
V. I. Kuznetsov ◽  
N. G. Gefan ◽  
N. M. Аndreevskaya ◽  
...  
Keyword(s):  
Nutrient Medium ◽  
Culture Medium ◽  
Specific Activity ◽  
Germination Rate ◽  
Biochemical Properties ◽  
Quantitative Composition ◽  
Nutrient Media ◽  
Microbiological Methods ◽  
Microbial Mass

Background. To obtain reliable results of laboratory studies on the identification of Listeria, the presence of certified diagnostic agglutinating Listeria sera is required. An important step in the manufacturing process of such medical devices for in vitro diagnostics requires effective nutrient media for the accumulation of listeriosis microbe. Aim of the research. To develop an effective nutrient medium for the accumulation of bacterial mass of Listeria. Materials and methods. The object of the study was an experimental culture medium for Listeria cultivation. As a control, we used nutrient agar for the cultivation of microorganisms (fish meal hydrolysate, FMH-agar) and meat-peptone agar with 1 % glucose (MPA with 1 % glucose). The specific activity of nutrient media during cultivation of the test strain Listeria monocytogenes 766 was evaluated using a complex of microbiological methods. Results. The optimal base of the nutrient medium for Listeria cultivation has been selected: pancreatic hydrolysate of river magpie fish (Rutilus rutilus lacustris) and hydrolysate of meat water production waste. The qualitative and quantitative composition of the nutrient medium has been developed, its physical, chemical and biological properties have been studied. It was found that after 24 hours of incubation at 37 °C, the nutrient medium provided the growth of typical Listeria colonies. The germination rate was 85 %, which is higher compared to the growth of the culture on MPA with 1 % glucose and GRM agar by an average of 21 % (p < 0,05). Conclusion. The experimental culture medium for Listeria cultivation provided growth of colonies of the test strain L. monocytogenes 766 with the preservation of characteristic cultural, morphological and biochemical properties, and, in terms of germination and growth rate, exceeded the control media. The developed nutrient medium provides effective growth of Listeria and can be used as a medium for the accumulation of microbial mass.

scholarly journals Regeneration capacity of Cerasus fruticosa and Prunus domestica into the in vitro culture

2021 ◽  
Vol 209 (06) ◽  
pp. 43-52
Author(s):  
Marina Markova ◽  
Elena Somova
Keyword(s):  
In Vitro Culture ◽  
Nutrient Medium ◽  
Optimal Time ◽  
In Vitro Cultivation ◽  
Initiation Time ◽  
Multiplication Factor ◽  
Optimal Conditions ◽  
Prunus Domestica ◽  
Nutrient Media

Abstract. The aim of these studies was to introduce into the in vitro culture the steppe cherry (Cerasus fruticosa) variety Shchedraya and the domestic plum (Prunus domestica) variety Sineokaya for subsequent micropropagation. Methods. Optimal conditions for obtaining viable explants, such as sterilizing agent and initiation time, have been investigated. The suitability of various nutrient media for in vitro cultivation of these cultures has also been tested. As a result of the experiments, it was revealed that the most effective sterilizing agents were 38 % perhydrol (control) and 6% chlorhexidine: the yield of viable cherry explants was 63.8 % and 61.5 %, plums – 69.8 % and 66.6 %, respectively. The optimal time for the initiation of cherry explants in vitro was January, where the yield of viable explants averaged 53.9 %, in June – 49.1 %, and for plums the initiation time did not matter – the yield of explants was 55.8 % in winter and 53.1 % in summer. In vitro cultivation of cherries and plums on the Quoirin – Lepoivre nutrient medium provided a significantly high multiplication factor, which averaged 4.1 for cherries (2.7 in control) and 6.0 for plums (3.9 in control). On the same medium, the maximum multiplication factor was obtained, which was 6.2 for cherries and 8.2 for plums. Thus, the scientific novelty of these studies is that the optimal conditions (sterilizing agent, time, nutrient medium) have been selected for the regeneration of cherry and plum explants in vitro with their subsequent micropropagation.

In vitro scaling of Tagetes patula L. root culture

2020 ◽  
Vol 19 (1) ◽  
pp. 255-259
Author(s):  
L. P. Khlebova ◽  
E. S. Brovko
Keyword(s):  
Growth Rate ◽  
Hairy Roots ◽  
Growth Pattern ◽  
Nutrient Medium ◽  
Root Culture ◽  
Growth Cycle ◽  
Tagetes Patula ◽  
Laboratory Conditions

The features of the in vitro accumulation of biomass of Tagetes patula L. root culture in the process ofscaling under laboratory conditions in different volumes of the nutrient medium and culture vessels were studied. It wasrevealed that the growth pattern of the hairy roots line and its morphology depend on the volume of the incubation vessel.The pattern of increasing the growth rate and lengthening of the growth cycle of the culture in larger flasks (up to 500 ml)was established.

scholarly journals Influence of various phytohormones on the growth and development of the Solnechny potato variety in vitro

2021 ◽  
Vol 36 ◽  
pp. 05008
Author(s):  
O.O. Novikov ◽  
M.S. Romanova ◽  
N.I. Leonov ◽  
E.I. Kosinova
Keyword(s):  
Surface Area ◽  
Potato Variety ◽  
Nutrient Medium ◽  
Growth And Development ◽  
Leaf Surface ◽  
Nutrient Media ◽  
Plant Weight ◽  
Total Leaf ◽  
Leaf Surface Area

In the work, the study of six different nutrient medium compositions’ influence on the growth and development of a healthy Solnechny potato variety in the growing process in laboratory conditions in vitro was carried out. The influence of different nutrient medium compositions on plant height, number of leaves/internodes, plant weight and total leaf surface area was studied. It was revealed that nutrient media with BAP addition cause a decrease in all of the above indicators. In addition, nutrient media with this growth regulator lead to the appearance of microtubers on the plants. It has been shown that the inclusion of GA and IAA growth regulators in nutrient media leads to a slight increase in the number of internodes. An increase in the total leaf surface area was noted in the above options. It should be noted that the nutrient medium options with the addition of GA and IAA led to a decrease in the mass and length of the root system of Solnechnyi variety potato plants.

scholarly journals Cultivation of the diatom algae Chaetoceros calcitrans f. pumilus (Paulsen) Takano, 1968 as food for giant oyster larvae Crassostrea gigas (Thunberg)

2019 ◽  
Vol 4 (2) ◽  
pp. 34-40 ◽  
Author(s):  
L. V. Ladygina ◽  
A. V. Pirkova
Keyword(s):  
Growth Rate ◽  
Crassostrea Gigas ◽  
Nutrient Medium ◽  
Growth Parameters ◽  
Inorganic Nitrogen ◽  
Diatom Algae ◽  
Nitrogen And Phosphorus ◽  
Nutrient Media ◽  
Chaetoceros Calcitrans ◽  
Giant Oyster

An impact of modified nutrient media F/2 and Conway on the growth and biomass accumulation of the diatom algae Chaetoceros calcitrans f. pumilus, which is a part of the food for cultivated larvae of the giant oyster Crassostrea gigas in the IMBR RAS nursery, was studied. Maximum values of cell and biomass concentrations were obtained on the modified F/2 nutrient medium (11.22 × 106 cells·ml-1 and 4.93 g·l-1, respectively), and they were much larger than those obtained on Conway medium. Growth parameters of C. calcitrans f. pumilus depended on the ratio of inorganic nitrogen and phosphorus, as well as on the silicon content in nutrient media. The ratio N : P = 12.5 and the silicon concentration of 24 mg·l-1 in the modified F/2 nutrient medium are shown to be approaching the optimal ones for increasing growth rate of diatom algae. It is found that the microalga in concentration 150 × 103 cells·ml-1, cultivated on different nutrient media and included in food composition, has impact on the growth rate of giant oyster larvae. An average daily amount of growth of larvae, whose diet included algae cultivated on modified F/2 nutrient medium, was higher than that of larvae cultivated on Conway medium.

scholarly journals INFLUENCE OF NUTRITION COMPOSITIONS ON MICROCLONAL PROPAGATION DIFFERENT GENOTYPES OF THE WALNUT JUGLANS REGIA L.

2020 ◽  
Vol 1 (383) ◽  
pp. 105-112
Author(s):  
T. K. Yegizbayeva ◽  
T. V. Yausheva ◽  
S. N. Oleichenko ◽  
R. J. Licea-Moreno
Keyword(s):  
Growth Rate ◽  
Nutrient Medium ◽  
Juglans Regia ◽  
Growth Dynamics ◽  
Initial Stage ◽  
Positive Growth ◽  
Murashige Skoog ◽  
Mother Plants ◽  
Quality Material

Walnut is one of the most common nuts in many countries of the world. It is used in cooking and in traditional medicine. Walnut cultivation on an industrial scale in Kazakhstan began only in 2016-2017, so Kazakhstan scientists in this field do not yet have any scientific experience. The article presents the results of studies on the influence of the nutrient medium compositions Murashige-Skoog and Driver-Kunzhuki on the microclonal propagation of different genotypes of the walnut Juglans regia L. Two varieties were selected as mother plants: the Uzbek variety Ideal and the Chinese variety Liaohe-1, adapted in the southeast Kazakhstan. In the course of work, we compared the growth rate of walnut shoots of two varieties on nutrient media Murasige-Skoog (MS) and Driver-Kunzhuki (DKW) with a concentration of 6-benzylaminopurine (BAP) of 1 and 1.5 mg / l. After a month of cultivation of shoots, positive growth dynamics was observed only on MS medium. On plants of both varieties, the growth rate was slow on DKW medium, the plants had yellow leaves, some shoots blackened and perished. To obtain high-quality material, as well as to increase the growth rate of shoots, DKW medium was supplemented with FeEDDHA (119 mg / L) and phloroglucinol (50 mg / L). As a result of phenological observations: the height of the main shoot, the number of internodes, the formation of additional shoots, and the state of the plants, it was found that the best nutrient medium for microclonal propagation of walnut Ideal and Liaohe-1 is a modified DKW medium. MS can only be used at the initial stage - an introduction to in vitro culture.

scholarly journals Influence of the component composition of nutrient media upon the introduction into the culture in vitro of explants of Prikubanskoe and Kubanskoe Bagryanoe apple varieties

2020 ◽  
pp. 18-23
Author(s):  
E. V. Lobodina ◽  
I. I. Suprun ◽  
A. O. Avakimyan ◽  
E. A. Al-Nakib
Keyword(s):  
Fungal Infection ◽  
Nutrient Medium ◽  
High Efficiency ◽  
Cumulative Effect ◽  
Component Composition ◽  
Apple Trees ◽  
Nutrient Media ◽  
Culture In Vitro ◽  
Apple Varieties

The work reflects the aspects of introducing two varieties of apple trees into the culture in vitro: Prikubanskoe and Kubanskoe Bagryanoe. The aims of study included testing a new disinfection protocol that accurately regulates the content of active chlorine in the disinfecting solution, as well as studying the regenerative ability of explants of apple trees of the Prikubanskoe and Kubanskoe Bagryanoe varieties on different culture medium: Murashige and Skoog (MS) ¼ NH4 NO3 , and Kvorin-Lepuavra (QL). The disinfection protocol used made it possible to obtain an insignificant level of explant contamination with fungal infection in passage zero, which indicates the effectiveness of the tested extraction protocol. The contamination of explants in passage zero by fungal infection was insignificant, which indicates the high efficiency of the sterilizing agent. The level of bacterial infection is higher in the Kubanskoe Bagryanoe variety (21.3 % of the total amount of explants). The influence of the component composition of nutrient media on the primary growth of apple micro-shoots was revealed. The following results were obtained: the yield of viable explants of the Prikubanskoe variety grown on nutrient medium MS ¼ NH4 NO3 was 44,4 %, which is 16,4 % higher than explants grown on QL medium (28 %). Explants of the Kuba nskoe bagryanoe variety, in turn, have a yield of vital microplants grown on MS ¼ NH4 NO3 16.7 %, which is 30.4 % lower than explants grown on QL medium (47.1 %). The combined effect of the genotype and the composition of the nutrient medium on the development of explants was statistically significant for the studied parameters. The share of the cumulative effect of factors of the variety x medium is 74.93 %, the influence of the genotype is 20.72 %. The data obtained indicate the variety specificity of the nutrient medium during clonal micropropagation of these apple varieties.

scholarly journals Significance of nutrient media choice for the long-term cultures of leukemic T-lymphoblasts

2021 ◽  
Vol 23 (3) ◽  
pp. 593-604
Author(s):  
L. S. Litvinova ◽  
K. A. Yurova ◽  
V. V. Shchupletsova ◽  
N. D. Gazatova ◽  
O. G. Khaziakhmatova ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Nutrient Medium ◽  
The Body ◽  
In Vitro Cultivation ◽  
Jurkat T Cells ◽  
Nutrient Media ◽  
Toxicological Studies

Correct choice of nutrient media for culturing different types of cells in various applications is one of the most important aspects of modern biotechnology, since chemical composition of the culture media largely contains the necessary metabolites to support certain cells’ growth lines outside the body. Jurkat line of human leukemic T-lymphoblast-like cells (hereinafter Jurkat T-cells) is actively used for in vitro modeling of intracellular signaling and activation of normal blood T-lymphocytes mediated by the T-cell receptor/CD3/ CD4 complex in toxicological studies of immune and secretory responses, to test medicinal substances and ions. Also, Jurkat T-cells are widely used for ex vivo testing in immunology, oncology, toxicology, orthopedics, and traumatology. The existing standards and numerous studies are mainly based on short-term in vitro cultivation of Jurkat T-cells in RPMI 1640 nutrient medium. Meanwhile, the issues of long-term maintenance of the growth of Jurkat T-cells culture are poorly presented in the research literature. This study aimed for studying the activity of Jurkat T-cells over 7 to 14 days of in vitro culture and comparing the relative value of RPMI 1640 and αMEM media for the behavior of immunocompetent tumor cells. Using flow cytometry, multiplex analysis, and phase contrast Cell-IQ microscopy, the proportions of living cells and those dying by apoptosis and necrosis, secretion of cytokines and chemokines, and the dynamics of cell biomass propagation were studied. It was found that the αMEM medium in the complete nutrient medium, as compared with RPMI 1640, is more appropriate to in vitro promotion of cell viability (increased proportion of viable cells by 13.5% at the day 14), their secretory ability for 23 из 27 tested biomolecules, shortened adaptation time (на 32%) in culture before growth initiation, 5-fold increase of the Jurkat Т-cell cellularity by the day 7. Potential significance of the chemical components of nutrient media and secreted biomolecules for these results is discussed. As based on the results obtained, we concluded on superior properties of αMEM medium for long-term in vitro cultures of Jurkat T-cells. Consequently, the in vitro testing of medical devices intended for long-term contact with the body, including those for cancer patients, using Jurkat T-cell leukemia line in RPMI 1640 medium, may lead to wrong predictions on their biocompatibility and potential antitumor activity.

scholarly journals REGENERATION CAPACITY OF NARROW-LOCALIZED ENDEMIC SPECIES Dianthus hypanicus Andrz. in vitro

2021 ◽  
Vol 14 (3) ◽  
pp. 39-45
Author(s):  
L.A. Koldar ◽  
Keyword(s):  
Growth Regulators ◽  
Nutrient Medium ◽  
Shoot Formation ◽  
Natural Habitats ◽  
Nutrient Media ◽  
Leafy Shoots ◽  
Mass Reproduction ◽  
Rare And Endangered ◽  
Propagation Methods

Development of effective technology of rare and endangered plant species mass reproduction in vitro is one of the urgent nowadays tasks. Diantus hypanicus Andrz. of the Caryophyllaceae Juss. family belongs to this group of plants. It is an endemic, narrow-localized species listed in the Red Book of Ukraine, the European Red List and the Annex to the Berne Convention. Prospects for the conservation of this species are associated with the study of its viability and require the development of effective reproduction methods. Aim. The purpose of the experiment was to determine the effect of different growth regulators concentrations on the D. hypanicus explants regenerative capacity during in vitro propagation. Methods. For the experiment the seed were used collected from plants of natural habitats and sown on a hormone-free nutrient medium according to the recipe of Murashige and Skoog. For 12–15 days, 4–6 cm long seedlings were obtained which were transferred to nutrient media modified with the content of growth-regulating substances: BAP 0.5–2.0 mg / l, IBA 0.01 mg / l and IAA 0.1 mg / l. Results. Initiation of adventitious buds with subsequent shoot formation was observed for 12–16 days. Each variant plants differed in number, growth activity and intensity, morphometric parameters. After 35–40 days from the explant introduction, conglomerates with well-developed leafy shoots were formed. It was found that, when modifying nutrient media with growth regulators BAP, IBA, IAA of different concentrations, D. hypanicus explants were actively undergoing regeneration processes of different intensity. Conclusions. It was proved that high ability to regenerate was the characteristic of explants cultured on a nutrient medium modified by the addition of BAP – 0.5 mg / l and IAA – 0.1 mg / l, where, on average, 18 ± 0.24 shoots of 2.42 ± 0.17 cm long were formed with a net reproduction of 19.06 ± 0.14. Shoots with 3.0–5.0 pairs of leaves that reached 3.5–6.0 cm were selected and transferred to nutrient media for rooting. Less developed shoots were planted on nutrient media for further reproduction. The next stage of the study is aimed at modifying nutrient media to achieve explants rhizogenesis and obtain a large number of plants necessary for the preservation of this endemic narrow-localized species in culture with subsequent repatriation to natural places of growth.

scholarly journals Comparative evaluation of hydrolysates as a basis for the construction of a nutrient medium for the cultivation of Listeria monocytogenes

2021 ◽  
Author(s):  
N. M. Khaptanova ◽  
А. S. Ostyak ◽  
S. V. Lukyanova ◽  
V. I. Kuznetsov ◽  
N. M. Аndreevskaya ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Enzymatic Hydrolysis ◽  
Nutrient Medium ◽  
Comparative Evaluation ◽  
Raw Materials ◽  
Amino Nitrogen ◽  
Test Strain ◽  
Nutrient Media ◽  
Microbiological Methods ◽  
The Common

The objective is to perform a comparative evaluation of the pancreatic hydrolysates prepared from fish and squid to determine the optimal culture medium for Listeria monocytogenes.Materials and methods. The following raw materials were used in the study: Pacific Herring (Clupea pallasii), Alaska Pollock (Gadus chalcogrammus), Common Roach (Rutilus rutilus lacustris), European Squid (Loligo vulgaris). The raw materials were subjected to enzymatic hydrolysis using the pancreas (according to Hottinger). A study of the physicochemical properties of pancreatic hydrolysates (content of free amino nitrogen (FAN), acidity of fish hydrolysates, the amino acid composition) was carried out.. The specific activity of nutrient media during the cultivation of the test strain L. monocytogenes 766 was assessed by a complex of microbiological methods.Results and discussion. The highest content of FAN at the end of enzymatic hydrolysis was observed in the pancreatic hydrolysate of the common roach (6%), the acidity of the hydrolysate remained stable from 6th to 13th day of the hydrolysis process (pH 7.2). Pancreatic hydrolysate of the common roach contained a number of amino acids that are most essential for the growth of Listeria. An assessment of the biological properties of nutrient media prepared on the basis of the obtained hydrolysates demonstrated that the best results in terms of sensitivity and germination of L. monocytogenes 766 showed a nutrient medium based on the pancreatic hydrolysate of the common roach. During the cultivation of L. monocytogenes 766 the test strain retained its morphological and cultural properties and did not show signs of dissociation.Conclusion. The research results have shown that the pancreatic hydrolysate of the common roach is a promising protein basis for the construction of an experimental environment for listeria. 

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