scholarly journals The obtaining potato microtubers on the basis of optimization of in vitro cultivation conditions

2021 ◽  
Vol 22 (5) ◽  
pp. 682-688
Author(s):  
E. N. Somova ◽  
M. G. Markova ◽  
E. A. Vlasevskaya
Keyword(s):  
Nutrient Medium ◽  
Sucrose Concentration ◽  
In Vitro Cultivation ◽  
Air Humidity ◽  
Cultivation Conditions ◽  
Potato Varieties ◽  
Relative Air Humidity ◽  
Early Ripening ◽  
A New Technique

Search work for optimization of such conditions for in vitro cultivation of potatoes as photoperiod, volume of nutrient medium, choice of a source of carbohydrate nutrition and its concentration, as well as growth regulators of auxin and cytokinin nature, was carried out in 2018-2020. Potato microplants of early-ripening (Alena, Latona, Red-Scarlett), middle-early (Adretta, Charodei, Svitanok Kievsky) and mid-season (Naiada, Ladozhskiy, Skarb) varieties were cultivated at illumination of 75-85 mMol/m2 s-1, 6500 K, air temperature 22...25 °C, relative air humidity 70-75 % and photoperiod from 4 to 16 hours. The results of three years of research have shown that the Murashige-Skooga nutrient medium modified by the Russian Potato Research Center with a 6 % sucrose concentration in a volume of 10 ml per microplant and a 12-hour photoperiod were optimal for micro-tuberization during in vitro cultivation of potatoes of all ripeness groups. The interaction of these cultivation conditions made it possible to obtain an average of 2.5 pcs. of microtubers per microplant of early-ripening potato varieties, 2.4 pcs. - middle-early and 3.2 pcs. - mid-season varieties. Optimal methods of in vitro cultivation of potatoes served as the basis for a new technique for obtaining potato microtubers. If this method was followed, the share of microplants with microtubers of early-ripening varieties increased by 6 %, middle-early varieties - by 12 % and mid-season ones - by 9 %. In addition, the duration of the micro-tuberization period in middle-early varieties was reduced by 14 days, in early-ripening and mid-season potato varieties by 28 days. Microplants of early-ripening and middle-early potato varieties formed larger microtubers, while mid-season varieties were in the lead in terms of quantitative yield.

scholarly journals Determination of xanthones in plants and the nutrient medium under in vitro cultivation conditions

2018 ◽  
Vol 76 (2) ◽  
pp. 33-37
Author(s):  
A. Revutska ◽  
V. Belava ◽  
A. Golubenko ◽  
N. Taran
Keyword(s):  
Nutrient Medium ◽  
Plant Material ◽  
In Vitro Cultivation ◽  
Cultivation Conditions ◽  
Quantitative Calculation ◽  
Environmentally Safe ◽  
Speed Up ◽  
Biotechnological Processes

In recent years, xanthones have received considerable attention from scientists due to their biological activity: anticarcinogenic, antiviral, antibacterial, antioxidant, anti-inflammatory and other properties.Therefore they are useful for prevention and treatment of different diseases:cancer, Alzheimer's and Parkinson's disease, cardiovascular disorders, diabetes, etc. Extracts of different species of plants containing xanthones are components of chemotherapeutic and other medical drugs. In order to find the most sensitive and environmentally safe method of quantitative determination of xanthones in the plant material and the nutrient medium, known methods were tested and selected for the prototype Vyisochina G. I. et al., 2011 method, which uses ethanol as an extractor. As the plant material we used plants of different species that were grown under in vitro cultivation conditions on the agarized nutrient medium. This agarized nutrient medium was also used for the xanthone content analysis. Based on the performed research, modifications of the method for determining the content of xanthones were adapted to the in vitro conditions, which detail the specificity of extraction and quantitative calculation of the xanthone content in plant explants. Our own method of determination of these compounds in the agarized nutrient medium was developed as well. The method, that we proposed, will significantly speed up the process of xanthone detecting and will also increase their yield in biotechnological processes for obtaining the pharmacologically valuable secondary metabolites of phenolic nature.

scholarly journals Optimization of in vitro cultivation conditions for rare and endangered species of Berberis iliensis and Berberis karkaralensis

2020 ◽  
Vol 132 (3) ◽  
pp. 43-56
Author(s):  
V.K. Karimova ◽  
◽  
B.N. Baktybai ◽  
G.K. Magzumova ◽  
ZH. T. Sartbaev ◽  
...  
Keyword(s):  
Endangered Species ◽  
Nutrient Medium ◽  
Anthropogenic Activities ◽  
In Vitro Cultivation ◽  
Cultivation Conditions ◽  
Rare And Endangered Species ◽  
Optimization Study ◽  
Rare And Endangered ◽  
Living Organisms

Today, many living organisms are negatively affected by climate change and anthropogenic activities, which leads to a decrease in their numbers. One of these rare and endangered plant species is the Ili barberry (Berberis iliensis) and the Karkaraly barberry (Berberis karkaralensis). This work is devoted to the optimization study of the cultivation conditions for a rare and endangered species of Ili barberry and Karkaralinsky barberry in vitro. To obtain sterile and viable explants, the sterilizing agent was a solution of 0.5% «Domestos» (7 min). For the regeneration of the main shoot of the barberry, the optimal nutrient medium is Murashige and Skoogwith the addition of 6-benzylaminopurine- 0.5 mg/l, gibberellic acid- 1.0 mg/l, indole-3-butyric acid -0.01 mg/l, where regeneration was 80% for the Ili barberry, barberry karkaralinsky - 70%. For the multiplication of Berberis iliensis microshoots, the Quoirin & Lepoivre culture medium with the addition of 0.75 mg/l - benzylaminopurine is optimal; the number of microshoots formed was 3.6 per explant. Root formation is one of the most difficult stages in micropropagation. For the rooting of microshoots of Karkaralinsky barberry, a nutrient medium of ½ Murashige and Skoog was used with the addition of indolylbutyric acid -1.5 mg/l.

scholarly journals INDUCTION OF MICROTUBING OF NEW PROMISING POTATO VARIETIES IN ASEPTIC CULTURE

2021 ◽  
Vol 16 (4) ◽  
pp. 48-54
Author(s):  
Elena Oves ◽  
Natal'ya Gaitova ◽  
Ol'ga Shishkina
Keyword(s):  
Nutrient Medium ◽  
Sucrose Concentration ◽  
Test Tube ◽  
High Yield ◽  
Stem Explants ◽  
Multiplication Factor ◽  
Potato Varieties ◽  
Aseptic Culture ◽  
Murashige Skoog

The studies were carried out with the aim of optimizing microtubing of promising potato varieties in aseptic culture. The experiments studied such factors as the use of vessels of various sizes (test tubes with a diameter of 25 mm and plastic containers 18x18 cm, into which 10 ml of agar and 400 ml of liquid nutrient medium, respectively, were poured, respectively), the density of planting plants in containers (40, 60 and 80 stem explants), the composition of the nutrient medium during ontogenesis, cultivation of mini-tubers from in vitro microplants (control) and microtubers (≥ 0.9 cm and 0.5 ... 0.9 cm in size). When studying the possibility of modifying the nutrient medium based on the Murashige-Skoog recipe to induce tuberization in one variant, the sucrose concentration during growth was changed from 2% before the formation of four internodes by 8% after this phase, the kinetin content during the entire observation period was 0.5 mg/l. In the second variant, microplants were kept on a medium with 6% sucrose and 0.25 mg/l kinetin throughout ontogenesis. The highest yield of microtubers of the standard fraction (27 ... 94%) with a multiplication factor of 0.8 ... 2.7 pcs/plant was noted in the variant with 60 cuttings placed in a container. When grown in test tubes with a change of medium, 1.0 ... 1.5 microtubers were collected per plant with a standard fraction yield of 64 ... 78%. The use of container technology with a similar alternation of nutrient media increased the yield of the standard fraction in most of the studied varieties to 75 ... 86%. In variants with a constant sucrose content in the nutrient medium (6%), a very low multiplication factor was noted, which did not compensate for a sufficiently high yield of the standard fraction, regardless of the laboratory vessel used. The multiplication factor of test tube microplants during planting in the ground was higher than when planting microtubers, with a high yield of the standard fraction

scholarly journals Effect of irradiation on the growth and rooting of a climbing rose in vitro

2021 ◽  
Vol 935 (1) ◽  
pp. 012007
Author(s):  
N Kondrateva ◽  
R Bolshin ◽  
M Krasnolutskaya ◽  
A Baturin ◽  
K Baturina ◽  
...  
Keyword(s):  
Surface Area ◽  
Leaf Surface ◽  
Air Humidity ◽  
Average Growth ◽  
Continuous Irradiation ◽  
Pulsed Mode ◽  
Relative Air Humidity ◽  
Leaf Surface Area ◽  
Reproduction Factor

Abstract The article presents the influence of pulsed and continuous irradiation (400…780 nm) on in vitro growth of the climbing rose variety “Camelot” at the illumination of 80±5 mmol/(m2s), temperature - 24±10C, and the relative air humidity - 73±2%. It was found that the experimental led light (LED) pulsed phytoirradiator contributed to a significant increase in the leaf surface area during cultivation of climbing rose microstems in Gamborg’s nutrient medium, the average growth was 2.94 mm2 compared to 2.80 mm2 in the control. Pulse irradiation increases the reproducibility of climbing roses by 1.7 times, and also increases the rooting rate up to 96% compared to 82% in the control. Experimental LED phytoirradiator of continuous irradiation promoted an increase in the leaf surface area growth at the level of the control luminescent phytoirradiator, but also provided a significant increase in the reproduction factor and rooting rate of rose microstems. In the pulsed mode, phytoinstallations consume only 50% of the electricity compared to the continuous irradiation mode.

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.

Influence of the Nutrient Medium and Photoperiod on Tuberization of Potato Micro-plants of Promising Potato Varieties in vitro Culture

2019 ◽  
Vol 5 (12) ◽  
pp. 177-181
Author(s):  
E. Vlasevskaya ◽  
I. Mukhametshin
Keyword(s):  
In Vitro Culture ◽  
Nutrient Medium ◽  
New Technologies ◽  
Sugar Concentration ◽  
Potato Seed ◽  
Potato Varieties ◽  
Federal Research ◽  
Production Technologies ◽  
Academy Of Sciences

The results of studies on the influence of the nutrient medium and photoperiod on tuberization of micro-plants of promising potato cultivars in an in vitro culture under are presented the conditions of a potato renewal laboratory at the Udmurt Research Institute of Agriculture of the Udmurt Federal Research Center of the Ural Branch of the Russian Academy of Sciences in 2018. Two experiments were carried out to identify patterns of the influence of potato cultivation conditions in vitro on the efficiency of its propagation and to develop an improved method for propagating potato micro-tubers in vitro. The experiments were carried out in accordance with the recommendation “New technologies for the production of healthy source material in elite potato seed production”, “Technologies for microclonal propagation of plants”. Object of research: micro-plants of potato varieties Alena, Charodei, Nayada. The studied variants of sugar concentration are 0%, 2% (control), 4%, 6%, 8%, 10%. The studied photoperiod options are 16 hours (control), 14 hours, 12 hours, 10 hours, 8 hours, 6 hours, 4 hours. Based on the results of studies in 2018, experimental data were obtained. It was revealed that the lack of sugar in the nutrient medium negatively affects the growth and development of potato micro-plants. An increase in sugar concentration to 6–10%, in comparison with the control (2%), significantly increases the tuber-forming ability of micro-plants by an average of 6.1–7.2% and increases the number of tubers from one micro-plant. The highest yield of micro-tubers from one micro-plant on average for varieties was obtained with a 14-hour photoperiod and amounted to 1.3 pcs. With a 12-hour photoperiod, micro-tubers begin to form 7–14 days earlier than in the other variants of the experiment.

scholarly journals Morphological peculiarites and functional activity of adipose-derived mesenchimal stem cells during in vitro cultivation conditions

2018 ◽  
Vol 20 (92) ◽  
pp. 79-82
Author(s):  
L. V. Kladnytska ◽  
A. I. Mazurkevych ◽  
V. T. Khomych ◽  
T. A. Mazurkevych ◽  
Z. G. Stegney ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Golgi Complex ◽  
Morphological Characteristics ◽  
Blue Dye ◽  
In Vitro Cultivation ◽  
Cultivation Conditions ◽  
Cytoplasmic Processes

The studies were conducted on 2-3-months-old males of C57BL/6 mice weighing 20–24 g. Obtaining and cultivating of adipose-derived mesenchimal stem cells (AD MSCs) were carried out in a sterile laminar box with compliance of conditions of asepsis and antiseptics. AD MSCs of the 2, 4, 7 and 12 passages were analyzed. Morphometric analysis was performed using a light microscopy. Morphometric parameters such as cell and nucleus area or nuclear-cytoplasmic ratio (NCR) were calculated using the Axiovision light microscope (Carl Zeiss, Germany) and ImageJ 1.45 software. Trypan blue dye used for investigation of the viability of MSC. The morphological characteristics of mesenchymal stem cells from adipose tissue during the process of cultivation changes: at the first passages of cultivation, the cells are spindle-shaped with two, at least three, long long cytoplasmic processes, located bipolar. Near the nucleus the Golgi complex is clearly visible – a sign of active cells. At later passages cells have a small cytoplasmic processes and the bipolar arrangement of processes changes by stellar arrangement. Golgi complex is also clearly visualized. The indicator of the nuclear-cytoplasmic ratio in MSC from adipose tissue is significantly reduced at 7 passage to 0.2189 ± 0.0122 (P < 0.01), and at 12 passage to 0.1111 ± 0.0086 (P < 0.001) compared to the 2 passage. The coefficient of proliferation of MSC from adipose tissue is significantly reduced at 12th passage. The viability of mesenchymal stem cells from adipose tissue with an increasing of a number of passages significantly reduces and at the 12th passage of cultivation reaches 84,67 ± 1,36* (P < 0.05). The content of apoptotic cells that exhibited sensitivity to serum-free significantly increased at 7 and 12 passages and was respectively 21.33 ± 1.36 (P < 0.05) and 23.67 ± 0.97% (P < 0.05).

scholarly journals Study of Phytophthora infestans Mont. de Bary isolates in the planting of potatoes

2021 ◽  
pp. 86-91
Author(s):  
N. V. Matsishina ◽  
P. V. Fisenko ◽  
O. A. Sobko ◽  
I. V. Kim ◽  
D. I. Volkov ◽  
...  
Keyword(s):  
Phytophthora Infestans ◽  
Molecular Genetic ◽  
Microscopic Analysis ◽  
Molecular Genetic Analysis ◽  
In Vitro Cultivation ◽  
Genetic Characteristics ◽  
Potato Varieties ◽  
Phytotoxic Activity ◽  
Wide Range

Relevance. One of the most common diseases of potatoes and other nightshade family species is late blight caused by a pathogenic oomycete of the Phytophthora infestans (Mont.) de Bary. At least 100 species of phytophthora have been described in nature, affecting a wide range of plant species. The phytophthora population is heterogeneous and is represented by races, as well as different types of mating. This leads to a rapid adaptation of the pathogen and the emergence of new, more aggressive, and resistant races. Phytophthora is a parasite, the damage from which cannot be avoided within the organic farming framework. Therefore, it is particularly important to know the pathogenesis and racial composition of phytophthora in each individual region of Solanaceae cultivation.Research methodology. Differentiation and collection of material from the natural population were carried out using potato varieties with known R-genes in the genome. Isolation and introduction into the culture were carried out from leaves with the dampening chambers method, followed by cultivation on nutrient media. The pathogen was identified by microscopic analysis. Culture filtrates were obtained on the liquid nutritious medium, followed by liquid filtration and autoclaving. Phytotoxic activity was determined by the effect on the seedlings of the nightshade, grass, and pea families by the standard method. Molecular genetic analysis of the isolates was carried out by ISSR analysis; the primer, amplification mixture, and temperature profile of the reaction were selected according to the literature data; the calculation of genetic characteristics was carried out using POPGENE software packages.Results. Samples of seven Phytophthora infestans isolates were collected and introduced into culture. As a result of in vitro cultivation, morphological differences were revealed, expressed in the structure and color of the mycelium, the shape of the colonies, the nature of sporulation, the color of the reverse, and the medium under the colonies. The genetic differences of the natural phytophthora material introduced into the culture, collected from potato varieties with single resistance genes (R1, R3, R4), were revealed. Differences in the phytotoxic activity of the studied isolates' cultural filtrates were revealed. The isolated isolates demonstrate differentiation at the phenotypic, genetic and physiological levels, which allows us to speak about their belonging to races.

scholarly journals A Journey into of the Universe of in vitro Cultures of Plants. Callogenesis

2019 ◽  
Vol 9 (4) ◽  
pp. 45
Author(s):  
Gogu Ghiorghita
Keyword(s):  
Nutrient Medium ◽  
In Vitro Cultures ◽  
Plant Tissues ◽  
Cultivation Conditions ◽  
In Vitro Morphogenesis ◽  
History Of ◽  
The Universe ◽  
Biological Nature

After presenting a brief history of the discovery of this system of cultivation of plant tissues and cells, of the evolution in time of knowledge in this field, of the pathways of in vitro morphogenesis in plants, the paper focuses on in vitro callogenesis in plants. There are presented the types of callus that can be generated via in vitro culture (illustrated with aspects from the author&#39;s experience), the influence of factors related to the biological nature of the explants, the composition of the nutrient medium, the cultivation conditions, etc., upon some characteristics of the callus obtained. A special chapter is dedicated to the importance of in vitro cultures of callus in plants.

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.

Sign in / Sign up

Export Citation Format

Share Document