scholarly journals In vitro rhizogenesis of sugar beet microclones

2017 ◽  
Vol 8 (4) ◽  
pp. 616-622
Author(s):  
V. V. Polishchuk ◽  
L. M. Karpuk ◽  
V. P. Mykolaiko ◽  
A. A. Polishchuk ◽  
I. I. Mykolaiko
Keyword(s):  
Sugar Beet ◽  
Chromosomal Abnormalities ◽  
Morphological Changes ◽  
Lateral Roots ◽  
Morphological Characteristics ◽  
In Vitro Cultivation ◽  
Male Sterile ◽  
Nutrient Media ◽  
Lower Efficiency

The features of rhizogenesis of male-sterile simple hybrids, maternal components of sugar beet heterozosis hybrid and O-types of Darinka variety grown from seeds were investigated. The paper presents the results of studying the main basic nutrient media (B5–A1, B5–A2, B5–A3, B5–A4, B5–A5 and B5–A6) for rhizogenesis, in which the content of macro- and microelements has been reduced by three times, the content of sucrose is reduced by almost 6 times, the amount of agar is reduced by almost five times, while the vitamin РР content is increased by 1.6 times, and the BAP is completely removed. The largest number of rooted microclones was obtained with NAAs by the high indexes of average number of roots and medium indexes of average roots length. To the second, by the number of rooted microclones is assigned, media of 2,4–D, 2,4–DB containing, potassium salt of NAA and IBA were classified, while IAA was characterized by a lower efficiency. The possibilities of regulation of growth and development processes of sugar beet in vitro explants in favor of undifferentiated growth at the stage of proliferation or organogenesis (hemo- and rhizogenesis) have been determined. To improve the quality of rooting of sugar beet plants-regenerants, we have developed the method for dimming agarized nutrient medium with methylene blue at a 0.05% concentration, which made it possible to reduce the inhibitory effect of light on the development of lateral roots. The average index of micro-roots rooting for MS-forms was 81.5 %. The technology of adaptation of micro-roots rooting was developed, in which the plant material at the initial stage of growth had slight morphological changes in the puffer apparatus and stem, but by the end of the vegetation, the plants acquired the appearance characteristic of the donor plants of explants. The clone’s resiliency is reached about 100% in the soil. According to the variability of morphological characteristics in reproduced in vitro sugar beet plants of the MS-forms of the Darynka hybrid, all the plants grown from the regenerants were more, than seed progeny from seed sowing of the same component obtained as a result of the attachment of sterility to the corresponding O-type. It has been found that in vitro cultivation has not weakened, and in some cases even increased the mitotic index of meristem cells of young roots of sugar beet. Most genotypes have reduced the number of pathological mitoses, apparently due to the activation of reparative systems, but did not extend the duration of individual phases of mitosis; without increasing the number of anaphase with bridges and fragments, but changed the number of anaphases with other chromosomal abnormalities. In all variants of the experiment, where activation of growth processes has been observed, the lowest variability of the signs is observed, that is, under the influence of stimulants, which are part of nutrient media, obviously there is unification of cell populations. 

scholarly journals Adaptability and leaf anatomical features in oil palm seedlings produced by embryo rescue and pre-germinated seeds

2010 ◽  
Vol 22 (3) ◽  
pp. 209-215 ◽  
Author(s):  
Zanderluce G. Luis ◽  
Kadja Milena G. Bezerra ◽  
Jonny Everson Scherwinski-Pereira
Keyword(s):  
Oil Palm ◽  
Embryo Rescue ◽  
Morphological Characteristics ◽  
Growth Conditions ◽  
Epidermal Cells ◽  
In Vitro Cultivation ◽  
Vascular Bundles ◽  
Anatomical Plasticity ◽  
Germinated Seeds

Changes in the leaf structure of plants grown in different conditions have been reported, such as increase in size and density of stomata and reduction in stomatal control, amount of epicuticular wax, and mesophyll thickness, with a high diversity of intercellular spaces. However, these changes are highly variable depending on the physiological and morphological characteristics of each species. The objective of this work was to analyze the adaptability and anatomical plasticity of oil palm seedlings produced after embryo rescue and pre-germinated seeds. Expanded leaves were prepared for evaluation of morphometric data and anatomical structures. It was verified that the environmental conditions in vitro negatively influenced the stomata density, epidermal and hypodermal thickness, and the values for the expansion cells and leaf mesophile. Anatomically, the oil palm leaves present the same tissues composition in both growth conditions, with uniseriate epidermal cells, and tetracitic stomata occurring in both epidermal surfaces. Epidermal cells from in vitro plants are thinner than ones from greenhouse. The midrib of leaves from greenhouse plants are more developed and is composed by only one central vascular bundle, while plants from in vitro cultivation developed three to four collateral vascular bundles.

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.

scholarly journals Development of the prickly pear cactus Opuntia stricta (Haw.) Haw. (Cactaceae) in vitro in response to the replacement of potassium nitrate for a commercial kno 3 fertilizer

2022 ◽  
Vol 52 (1) ◽  
Author(s):  
Cinthia Carolinne de Souza Ferreira ◽  
Cristiane Domingos da Paz ◽  
Joselita Cardoso de Souza ◽  
Ana Rosa Peixoto ◽  
Lucas Silva Rios ◽  
...  
Keyword(s):  
Morphological Changes ◽  
Potassium Nitrate ◽  
Prickly Pear ◽  
In Vitro Cultivation ◽  
Control Medium ◽  
Fresh Biomass ◽  
Prickly Pear Cactus ◽  
Product Costs ◽  
Pear Cactus

ABSTRACT: In micropropagation, potassium nitrate (KNO3), an ACS reagent grade chemical, used in the preparation of growing mediums is expensive and its procurement depends on bureaucratic procedures, as it is controlled by the Brazilian Army. This research to assessed the effect of replacing the ACS KNO3 for a commercially available fertilizer (KNO3- based) on the micropropagation of the prickly pear cactus (Opuntia stricta (Haw.) Haw. cv. Elephant Ear. Treatments used six different fertilizer concentrations (0, 0.5, 1, 1.5, 2 and 2.5 g L-1) and a control consisting of 1.9 g L-1 KNO3, as shown in the MS salts. The survival, size and number of sprouts and the value of fresh biomass were evaluated. After seedling acclimation, we assessed the survival, number of sprouts, length, and number of roots, racket formation, average fresh biomass mass, macronutrient absorption and morphological changes of the seedlings. Explants inoculated with fertilizers at concentrations of 0.0; 2.0 and 2.5 g L-¹ did not grow. The response of explants at concentrations of 0.5 and 1.5 g L-1 of the fertilizer were the same as those developed in a KNO3 medium, and at a concentration of 1.0 g L-1, in all variables, the means were higher than those of the control medium. Therefore, it showed the feasibility of using fertilizers in the in vitro cultivation of the prickly pear cactus, which may remove bureaucratic barriers and reduce product costs by 99.12%.

scholarly journals ФЕНОТИПОВІ ТА МОРФОЛОГІЧНІ ЗМІНИ КУЛЬТУРИ КЛІТИН КІСТКОВОГО МОЗКУ ЩУРІВ В ПРОЦЕСІ ЇХ КУЛЬТИВУВАННЯ

2016 ◽  
Vol 18 (2(66)) ◽  
pp. 126-132
Author(s):  
A.I. Mazurkiewicz ◽  
V.V. Kovpak ◽  
O.S. Kovpak
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Morphological Changes ◽  
Embryonic Stem ◽  
In Vitro Cultivation ◽  
Rat Bone ◽  
Marrow Cells

Bone marrow is the only adult tissue which normally consists of immature undifferentiated and low differentiated cells which called stem cells and they are similar in structure to embryonic stem cells. But literature data analysis doesn't give an unambiguous answer regarding phenotypic and morphological changes of bone marrow cells culture of rats during their in vitro cultivation which necessitated further research.Investigate phenotypic and morphological changes of bone marrow cells culture of rats during their in vitro cultivation from first to fourth passage.We were used in these research bone marrow cells of rats from the first to the fourth passages. Microscopic analysis and evaluation morphological changes of bone marrow cells culture of rats during cultivation were carried out using inverted microscope Axiovert 40. Control of changes phenotype was performed by detecting CD markers (CD10, CD38, CD34, CD45, CD48, CD54, CD56, CD66e, CD96, CD227, CD326, pan–keratin). The evaluation was performed by the semi– quantitative method (H–Score).The research of primary culture of rat bone marrow cells showed that it morphologically heterogeneous, noted the small number of cells polygonal shape, surrounded by the fibroblast cells. During the cultivation cell culture becomes more homogenous at the expense of fibroblast–like cells. As a result of occurred the transition process from heterogeneous culture in zero passage to the most homogeneous culture in 4 passage. Immunophenotyping population of cell culture derived from rat bone marrow, revealed a high level of expression of pan–keratin; moderate level – CD34, CD48, CD66e, CD95; low level – CD38, CD45, CD56, CD227, CD326; lack of expression – CD10, CD54. Change of the expression of surface markers varies in each passage CD48, CD66e, CD95 increased significantly; CD38, SD45, SD326, pan–keratin reduced significantly. The markers CD34, CD 56, CD 227 were expressed on the one level from the first to the fourth passage. The expression of the CD10, CD54 markers during the study period was not identified.

In vitro production of horse embryos predisposes to micronucleus formation, whereas time to blastocyst formation affects likelihood of pregnancy

2019 ◽  
Vol 31 (12) ◽  
pp. 1830 ◽  
Author(s):  
Kaatje D. Ducheyne ◽  
Marilena Rizzo ◽  
Juan Cuervo-Arango ◽  
Anthony Claes ◽  
Peter F. Daels ◽  
...  
Keyword(s):  
Chromosomal Abnormalities ◽  
Morphological Characteristics ◽  
Blastocyst Stage ◽  
Culture Conditions ◽  
Early Embryo ◽  
Embryo Morphology ◽  
In Vitro Production ◽  
Pregnancy Failure ◽  
Micronucleus Formation

Invitro embryo production is an increasingly popular means of breeding horses. However, success is limited by a high incidence of early embryo loss. Although there are various possible causes of pregnancy failure, chromosomal abnormalities, including aneuploidy, are important potential contributors. This study evaluated the frequency of micronucleus formation as a proxy for aneuploidy in invitro-produced (IVP) and invivo-derived horse blastocysts. Associations between IVP embryo morphology, frequency of nuclear abnormalities and the likelihood of pregnancy were investigated. IVP blastocysts exhibited a higher frequency of cells with micronuclei than invivo-derived embryos (10% vs 1% respectively; P=0.05). This indication of chromosomal instability may explain the higher incidence of pregnancy failure after transfer of IVP embryos. However, the frequency of micronuclei was not correlated with brightfield microscopic morphological characteristics. Nevertheless, IVP embryos reaching the blastocyst stage after Day 9 of invitro culture were less likely to yield a pregnancy than embryos that developed to blastocysts before Day 9 (27% vs 69%), and embryos that had expanded before transfer were more likely to undergo embryonic death than those that had not expanded (44% vs 10%). These findings indicate that current embryo culture conditions are suboptimal and that the speed of embryo development is correlated with pregnancy survival.

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 Modern issues of sugar beet (Beta vulgaris L.) hybrid breeding

2021 ◽  
Vol 25 (4) ◽  
pp. 394-400
Author(s):  
S. D. Karakotov ◽  
I. V. Apasov ◽  
A. A. Nalbandyan ◽  
E. N. Vasilchenko ◽  
T. P. Fedulova
Keyword(s):  
Mitochondrial Genome ◽  
Sugar Beet ◽  
High Performance ◽  
High Efficiency ◽  
Practical Importance ◽  
Hybrid Breeding ◽  
Haploid Plant ◽  
Male Sterile ◽  
Rapd Primer

High efficiency of the cultivation of unfertilized sugar beet ovules and preparation of haploid regenerants (microclones) of pollinators – maintainers of О-type sterility and MS forms of the RMS 120 hybrid components has been shown. A technological method that accelerates the creation of new uniform starting material is proposed. It speeds up the breeding process two to threefold. The identification of haploid regenerants with sterile cytoplasm in initial populations is of great theoretical and practical importance for breeding, as it facilitates the production of homozygous lines with cytoplasmic male sterility and high-performance hybrids on sterile basis. As shown by molecular analysis, a single-nucleotide polymorphism never reported hitherto is present in the mitochondrial genome of the haploid plant regenerants. It allows identification of microclones as fertile and sterile forms. It has been found that DNA markers of the sugar beet mitochondrial genome belonging to the TR minisatellite family (TR1 and TR3) enable reliable enough identification of haploid microclonal plants as MSor O-type forms. Fragments of 1000 bp in length have been detected in monogenic forms in the analysis of 11 sugar beet plants cultured in vitro by PCR with the OP-S4 random RAPD primer. Testing of the OP-S4 marker’s being in the same linkage group as the genes responsible for expression of the economically valuable trait monogermity demonstrates its relative reliability. By the proposed method, dihaploid lines (DH) of the male-sterile form and the О-type sterility maintainer of the RMS 120 sugar beet hybrid have been obtained in in vitro culture. These lines are highly uniform in biomorphological traits, as proven under field conditions.

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 In vitro ectomycorrhiza formation by monokaryotic and dikaryotic isolates of Pisolithus microcarpus in Eucalyptus grandis

2010 ◽  
Vol 34 (3) ◽  
pp. 377-387 ◽  
Author(s):  
Maurício Dutra Costa ◽  
André Narvaes da Rocha Campos ◽  
Matheus Loureiro Santos ◽  
Arnaldo Chaer Borges
Keyword(s):  
Lateral Root ◽  
Morphological Changes ◽  
Lateral Roots ◽  
Eucalyptus Grandis ◽  
Model Systems ◽  
Root Yield ◽  
Root Tips ◽  
In Vitro Synthesis ◽  
Synthesis Medium

The formation of ectomycorrhizas by monokaryotic and dikaryotic isolates of Pisolithus microcarpus (Cooke & Massee) G. Cunn. in Eucalyptus grandis W. Hill ex Maid. was studied by in vitro synthesis in Petri dishes. The formation of ectomycorrhizas was observed for all strains tested. Ectomycorrhizas formed by the monokaryotic strains presented a sheath of hyphae around the roots and a Hartig net limited to the epidermis layer, typical of the angiosperm ectomycorrhizas. Colonization rates, a measure of the number of ectomycorrhizas in relation to the total number of lateral root tips, varied from 23 to 62%. Some monokaryotic strains stimulated the formation of lateral roots, promoting increases of up to 109% above the control. The presence of some of the isolates in the in vitro synthesis medium stimulated the production of thicker lateral root tips. The dimensions of the lateral roots tips and ectomycorrhizas varied from one isolate to the next, indicating a variation in their capacity to provoke morphological changes in the host plant roots. The dikaryotic strain M5M11 presented higher values for lateral root yield, number of ectomycorrhizas, and colonization percentage than the corresponding monokaryotic strains, M5 and M11. This indicated the possibility of selecting compatible performing monokaryotic isolates for the yield of superior dikaryotic strains. The set of monokaryotic strains tested varied greatly in their ability to colonize E. grandis roots and cause secondary metabolism-related morphological changes in roots, providing a wealth of model systems for the study of genetic, physiological, and morphogenetic processes involved in Pisolithus-Eucalyptus ectomycorrhiza formation.

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