scholarly journals Evaluation of the effectiveness of potato improvement in in vitro culture on optimized nutrient media

2021 ◽  
Vol 36 ◽  
pp. 05007
Author(s):  
I.A. Gneusheva ◽  
A.Yu. Gavrilova ◽  
I.N. Gagarina ◽  
I.V. Gorkova ◽  
I.Yu. Solokhina
Keyword(s):  
Potato Variety ◽  
Nutrient Medium ◽  
Antiviral Drug ◽  
Antimicrobial Properties ◽  
Test Tube ◽  
Nutrient Media ◽  
Active Growth ◽  
Potato Improvement ◽  
Trichoderma Lixii

Studies have been conducted on the use of an optimized Murashige-Scoog nutrient medium to obtain healthy seeds of promising potato varieties. It was found that the use of agarized modified Murashige-Scoog nutrient medium with the addition of exometabolites of fungi of the genus Trichoderma lixii 1.5 mg/l with antimicrobial properties and the antiviral drug “Cycloferon” in a concentration of 0.01% contributes to an increase in the number of virus-free potato regenerants when the method of apical meristems and in vitro improvement is used together. The effectiveness of using the modified medium is confirmed by the following criteria: active growth of test tube potato micro-plants, which is expressed in optimal morphometric indicators on the example of the potato variety “Rosara”; the size of the isolated meristems is 10 mm, which contributes to the output of healthy plants; the survival rate of regenerating plants. It is shown that when using the modified medium, an average of 35-50% of regenerating plants were obtained, depending on the potato variety, free from M, S viruses.

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 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.

REGENERATION OF APPLE VARIETIES IN VITRO CULTURE

2020 ◽  
pp. 22-25
Author(s):  
L. V. Tashmatova ◽  
O. V. Matsneva ◽  
T. М. Khromova ◽  
V. V. Shakhov
Keyword(s):  
Growth Phase ◽  
Proliferative Activity ◽  
Small Degree ◽  
Nutrient Media ◽  
Regenerative Capacity ◽  
Active Growth ◽  
Apple Varieties ◽  
Clonal Micropropagation ◽  
Genotypic Characteristics

The process of clonal micropropagation is influenced by the genotypic characteristics of plants, the composition of nutrient media and the hormonal background at various stages of cultivation. The purpose of the research was to study the regenerative capacity of apple explants of promising varieties. The objects were colon-shaped, immune and scab-resistant apple varieties. The study was carried out in the laboratory of biotechnology of VNIISPK using generally accepted methods. The largest number of viable explants was obtained during the active growth phase, averaging from 72.1 to 88.5%. Maximum contamination was observed in explants introduced in April, regardless of the sterilizing agents. The highest infection rate was in the varieties Orlovskoe рolesie (up to 56.7%), Bolotovskoe (up to 51.4%), Poeziya (up to 43.3%). In the explants introduced in June, the infection was insignificant. Thus, the Imrus and Poeziya varieties showed a small degree of infection after using 0.1% mertiolate solution (1.2% and 2.4%), and the garland and Priokskoe varieties had a 100% yield of pure apexes. Among immune varieties, the highest necrosis was evident in the variety of Imrus (42,1%) and the smallest in the variety Kandil оrlovsky (5,1%) have the largest column-the variety Priokskoe (65.6 per cent) and Vostorg (65,6%). In most of the studied varieties, the greatest necrosis was observed after using 0.1% mertiolate solution. The study was carried out on the background medium QL and Fardzinova recommended for pears. Concentration of BAP 1.0 and 2.0 mg/l. Among the immune varieties, the content of BAP 2.0 mg / l in the QL medium caused the greatest proliferative activity in the varieties Kandil оrlovsky (3.4), Bolotovskoe (4.5) and Orlovskoe рolesie (2.9). Varieties of Imrus and a Veteran of the highest rate of multiplication were obtained in Fardzinova on medium containing BAP in both concentrations. In all varieties, the concentration of BAP 1.0 mg/l contributed to the formation of shoots more than 1 cm, i.e. suitable for rooting.

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 Use of growth inhibitor chlormequat chloride in potato culture in vitro

2021 ◽  
Vol 285 ◽  
pp. 03003
Author(s):  
Sergey A. Buldakov
Keyword(s):  
Nutrient Medium ◽  
Growth Inhibitor ◽  
Test Tube ◽  
Investigational Product ◽  
Chlormequat Chloride ◽  
Skoog Medium ◽  
Potato Culture ◽  
Murashige And Skoog Medium ◽  
The Cost

The publication presents data on the use of growth inhibitor chlormequat chloride in in vitro potato culture on varieties of different maturity groups: Meteor (early), Zekura (mid-early), and Northern Lights (mid-season). Five dosages of the investigational product were studied, ranging from 0.1125 to 1.8 g/l. It was found that in maximum doses of the product there was a strong inhibition of all growth processes in all varieties. The research results showed that the most optimal concentration of chlormequat chloride is 0.225 g per 1 liter of Murashige and Skoog medium. At this dose, on 30th day of cultivation, there was a decrease in the height of microplants from the control by 63.2-85.1%, in the root length - up to 15.0% and their number - up to 22.8% and an increase in the number of internodes by 6.5-22,0 % depending on the variety. The investigational product had an effect on formation of microtubers; in the Meteor variety, their largest number was 89.5% in the nutrient medium with a dose of 1.8 g/l, in the Zekura variety - 93.0% in the nutrient medium with a dose of 0.9 g/l. The new technique makes it possible to lengthen the periods between cuttings of test-tube plants by 2.3 times. This, in turn, reduces the cost of maintaining the in vitro collection material of potatoes in the summer-autumn period, and improves its quality, since each additional cutting cycle affects a more rapid degeneration of the variety. Also, microplants grown with growth inhibitor during subsequent relocation to a standard Murashige and Skoog medium did not show an aftereffect.

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 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 The study of the method of culture in vitro as a method of vegetative propagation of coniferous trees

2019 ◽  
pp. 19-22
Author(s):  
V. V. Krasnoperova ◽  
I. L. Bucharina
Keyword(s):  
Growth Regulators ◽  
Vegetative Propagation ◽  
Test Tube ◽  
Dichlorophenoxyacetic Acid ◽  
Nutrient Media ◽  
Culture In Vitro ◽  
Coniferous Trees ◽  
Planting Material ◽  
Economically Valuable Traits

Research is aimed at improving the technology of vegetative propagation of coniferous trees in vitro by selecting optimal media for sterilizing the explants of vegetative parts of plants, and using growth regulators to obtain callus and stimulating the root formation of test-tube plants. The objects of study are the vegetative parts of conifers: the buds and parts of the annual shoots. Plant objects sterilized and planted on nutrient media of different composition according to the scheme of experiments. Laboratory studies were conducted, on the basis of which successful use of sodium hypochlorite was noted compared with other reagents for sterilization of explants. According to the results of the experiments, a Woody Plant Medium nutrient medium was identified which promotes the best survival of the explants of conifers. The effect on growth survival of explants and their viability in culture in vitro, growth regulators such as 2.4-dichlorophenoxyacetic acid and 2.4-dinitrophenol in nutrient media was also determined. With the further cultivation and reproduction of coniferous plants, this technique allows to obtain planting material that most economically preserved in the offspring the economically valuable traits and properties of the parent plant.

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