scholarly journals Modeling of asymmetric division of somatic cell in protoplasts culture of higher plants

2020 ◽  
Vol 11 (2) ◽  
pp. 255-265
Author(s):  
S. I. Kondratenko ◽  
T. P. Pasternak ◽  
O. P. Samovol ◽  
O. M. Mogilna ◽  
O. V. Sergienko
Keyword(s):  
Cell Division ◽  
Osmotic Stress ◽  
Growth Regulators ◽  
Nutrient Medium ◽  
Higher Plants ◽  
Asymmetric Division ◽  
Cycle Phase ◽  
In Planta ◽  
Selection Of

The key result of the work is the selection of factors for the cultivation of protoplasts of higher plants in vitro, which allowed induction of asymmetrical cell division during the first cell cycle phase. Gibberellin has been proved to be one of the main cofactors of asymmetric division of plant cells. The objects of research were plants of the following cultivars aseptically grown in hormone-free MS medium: tobacco (Nicotiana tabacum L.), SR-1 line; Arabidopsis thaliana var. columbia (L.) Heynh; potato (Solanum tuberosum L.), Zarevo cultivar; cultivated white head cabbage (Brassica oleraceae var. capitata L.) of the following varieties: Kharkivska zymnia, Ukrainska osin, Yaroslavna, Lika, Lesya, Bilosnizhka, Dithmarscher Früher, Iyunskarannya; rape (Brassica napus L.) of Shpat cultivar; winter radish (Raphanus sativus L.) of Odessa-5 cultivar. In experiments with mesophilic and hypocotyl protoplasts of the above-mentioned plant species it has been proved that short-term osmotic stress within 16–18 hours being combined with subsequent introduction of high doses of gibberellin GK3 (1 mg/L) into the modified liquid nutrient media TM and SW led to the occurrence of pronounced morphological traits of cytodifferentiation already at the initial stages of the development of mitotically active cells in a number of higher plants. Meanwhile, in all analyzed species, there was observed the division of the initial genetically homogeneous population of mitotically active cells into two types of asymmetric division: by the type of division of the mother cell into smaller daughter cells and by the type of the first asymmetric division of the zygotic embryo in planta. In this case, the first type of asymmetric division occurred during unusual cytomorphism of the mother cells: a pronounced heart-shaped form even before the first division, which is inherent in the morphology of somatic plant embryo in vitro at the heart-shaped stage. A particular study of the effect of osmotic stress influencing protoplasts of various cultivars of white cabbage, isolated from hypocotyls of 7–9 day etiolated seedlings, revealed quite a typical consistent pattern: the acquisition and maintenance of the axis of symmetry in growing microcolonies occurred without extra exogenous gibberellin (GK3), which was added to the nutrient medium earlier. While analyzing the effect of growth regulators on the formation of microcolonies with traits of structural organization, the conclusion was made regarding the commonality of the revealed morphogenetic reactions of cells within the culture of protoplasts of higher plants in vitro with similar reactions studied earlier on other plants, both in vitro and in planta. Modeling of asymmetric cell division in protoplast culture in vitro has become possible by carrying out a balanced selection of growth regulators as well as their coordinated application through time along with changes in osmotic pressure of a nutrient medium.

scholarly journals Multi-Step Cellular and Tissue Selection of Soybean for Resistance to Osmotic Stress Using Peg 6000 Invitro Conditions

2021 ◽  
Vol 8 (4) ◽  
Author(s):  
Mohammad Naim Noori
Keyword(s):  
Polyethylene Glycol ◽  
Osmotic Stress ◽  
Nutrient Medium ◽  
Selective Agent ◽  
Soybean Variety ◽  
Peg 6000 ◽  
Final Volume ◽  
Polyethylene Glycol 6000 ◽  
Selection Of

Cell and tissue selection in vitro allows targeted selection of the desired traits under severe selective conditions at the level of individual cells and tissues. On the basis of multistage cell and tissue selection with the use of a selective agent - neutral osmotic polyethylene glycol 6000 in increasing concentrations -5%, 10%, 15%, 20% of the final volume of the nutrient medium, callus of the Zhansaya soybean variety that are stably resistant to osmotic stress have been obtained. Regeneration from callus was noted only in8.3 % of the planted callus 6 soybean regenerants plants resistant to osmotic stress were obtained from callus.

scholarly journals Introduction to in vitro culture and callus initiation in Salvia hispanica L. (chia)

2019 ◽  
Vol 17 (1) ◽  
pp. 33-37
Author(s):  
A. Z. Revutskaya ◽  
A. V. Holubenko ◽  
N. V. Nuzhyna ◽  
H. O. Rudik ◽  
N. Yu. Taran
Keyword(s):  
In Vitro Culture ◽  
Growth Regulators ◽  
Nutrient Medium ◽  
Callus Growth ◽  
Basic Medium ◽  
High Concentration ◽  
Salvia Hispanica ◽  
Callus Initiation ◽  
Salvia Hispanica L

Aim. Preparation of aseptic seedlings Salvia hispanica L., callus initiation in vitro and establishment of primary explants suitable for the callus production. Methods. Seeds are sprouted on our own modification of conventional methods. The non-hormonal Murashige-Skoog agarized nutrient medium was used as basic medium for the experiments. Parts of one-month seedlings (roots, hypocotyl, cotyledon leaves) were used as explants for the use of the colza. We added growth regulators (BAP, 2,4-D) in different concentration combinations into the nutrient medium for callus initiation. Statistical processing was performed in Microsoft Office Excel. Results. Aseptic S. hispanica seedlings have been obtained. The callus growth was initiated on all types of explants, the dependence of the callus intensity on the type of explants and the growth regulators content in the nutrient medium was established. Morphogenic callus and root-regenerants have been obtained. Conclusions. Hypocotyl was the most suitable primary explant for callus growth. Seedlings, leaves and roots showed low morphogenetic capacity. The nutrient medium with an elevated 2,4-D content was the most effective for initiation of callus genesis and proliferation of non-morphogenous callus. A high concentration of 2,4-D in the medium improves S. hispanica callus growth but suppresses its morphogenic ability.Keywords: Salvia hispanica (Chia), in vitro culture, callus.

scholarly journals Choosing reference genes for RT-qPCR for Fusarium graminearum plant infection (In Planta) and In Vitro growth studies based on transcriptomic data

2019 ◽  
Author(s):  
Xiaorong Lin ◽  
Hongchen Li ◽  
Zonghua Wang ◽  
Stefan Olsson
Keyword(s):  
Fusarium Graminearum ◽  
Reference Genes ◽  
Target Genes ◽  
Housekeeping Genes ◽  
Relative Difference ◽  
Transcriptome Data ◽  
In Planta ◽  
Plant Infection ◽  
Selection Of

Background. Choosing reference genes for RT-qPCR for the study of transcriptomic responses of target genes is often done using “standard” reference genes (housekeeping genes) selected before the genomic era. Now, published transcriptome data can be used to aid in this selection to avoid the selection of a reference gene that varies and obscure results. Methods. We use transcriptome data for the model pathogen fungus Fusarium graminearum to select housekeeping genes for In Vitro and In Planta conditions. Transcriptome data was downloaded from a publicly available database. We selected a database where transcriptome chip data from many experiments using the same chip has been deposited divided the downloaded data into In Vitro and In Planta conditions based on the information about the experiments. Results. We ranked the genes with the least variation (relative difference between maximum and minimum expression) across each dataset. Genes previously shown to perform well as reference genes for In Vitro conditions in a similar analysis as ours also performed well for In Vitro conditions in our dataset but worked less well for In Planta conditions. We found 5 reference genes that performed well under both In Planta conditions and In Vitro conditions. Discussion. Even if these 5 reference genes performed well, for other (new) target conditions we recommend making a transcriptome analysis to select well performing reference genes for RT-qPCR if possible. Alternatively, select 2 of the 5 genes that we show here performed well under both In Planta and In Vitro conditions.

scholarly journals INTRODUCTION OF EXPLANTS AND REPRODUCTION ON NUTRIENT MEDIUM OF DONOR MATERIAL IN VITRO VARIETIES OF CALLISTEPHUS CHINENSIS (L.) NESS. FOR ITS FURTHER USE IN LANDSCAPING

2020 ◽  
Vol 1 (383) ◽  
pp. 89-96
Author(s):  
V. Polishchuk ◽  
S. Turchina ◽  
A. Balabak ◽  
I. Kozachenko ◽  
V. Mamchur ◽  
...  
Keyword(s):  
Plant Breeding ◽  
Vegetative Propagation ◽  
Nutrient Medium ◽  
Cell Biology ◽  
Plant Biotechnology ◽  
Forest Steppe ◽  
Physical Parameters ◽  
Adaptation Period ◽  
Selection Of

The relevance of the research topic. On the recent methods of biotechnology are increasingly used in plant breeding and seed production. Herbaceous plants such as strawberries, potatoes, a vegetable, some medicinal and others are capable of vegetative propagation the traditional methods of culture, successfully introduced in both in vitro and can achieve a high rate of reproduction. Modern plant biotechnology – the sum of the technologies developed in molecular and cell biology of plants – a new stage in the development of the technology of plant breeding. With these improved characteristics may occur at the level of individual genes and individual genes that determine a specific trait, can be identified. They may be the final selection, they can be isolated, insert, delete, or modify the genotype or variety. Goal. Identify the features of the manifestation of economically valuable features and decorative properties of Callistephus chinensis and the inclusion of the best varieties in the biotech link, their adaptation to the conditions of the Forest-Steppe of Ukraine and their further use in landscaping. Methods. Laboratory – determination of seed germination; mathematical and statistical - for processing the reliability of the obtained research results. Results. The nutrient medium for growing plant tissues and cells, by analogy with the medium for culturing animal tissues, should contain all that the tissues in the plant organism receive from xylem and phloem currents of substances. However, in practice it has been found that vegetable juices cannot serve as a complete nutrient medium for growing isolated tissues and cells. This manifests the specificity of the receipt, transportation and especially the redistribution of nutrients in the plant. Based on the analysis, research was conducted to study the possibility of mass off-season vegetative propagation of plants of Callistephus chinensis in vitro. Practical recommendations on the selection of sterilizer, sterilization, nutrient medium and for the adaptation period of the best genotypes of this culture have been developed. As a result of the conducted researches the methods of selection of the initial plant material of Callistephus chinensis (Callistephus Chinensis (L.) NEES) and its surface sterilization, modification of existing aseptic culture methods have been studied and mastered. The morphogenetic potential of explants from different plant organs was investigated and selection of nutrient medium and study of the influence of plant growth regulators and physical parameters on the process of morphogenesis was carried out. The features of regeneration of isolated explants depending on the composition of the nutrient medium and selection of conditions for obtaining self-clones of Callistephus chinensis (Callistephus Chinensis (L.) NEES) were studied. Key words: in vitro, plant biotechnology, Callistephus Chinensis, nutrient medium, rhizogenesis.

scholarly journals Establishment of glyphosate selective concentrations for winter rapeseed (Brassica napus L.) transgenic tissues efficient selection in vitro

2019 ◽  
Vol 25 ◽  
pp. 197-201
Author(s):  
I. S. Hnatiuk ◽  
O. I. Varchenko ◽  
M. F. Parii ◽  
Yu. V. Symonenko
Keyword(s):  
Brassica Napus ◽  
Nutrient Medium ◽  
In Vitro Selection ◽  
Brassica Napus L ◽  
Regeneration Stage ◽  
Winter Rapeseed ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Efficient Selection ◽  
Selection Of

Aim. To establish the optimal concentration of glyphosate for efficient selection of transgenic tissue culture in biotechnology of rapeseed by in vitro selection of glyphosate-resistant and non-resistant winter rape. Methods. As an explants 7–10 mm fragments of 6-day rapeseed hypocotyls were used to cultivate on the medium, supplemented with 1 mg/L of 2,4-dichlorophenoxyacetic acid (2,4-D) for 12 days at 24°C under dark conditions for the initiation of callusogenesis. For regeneration of plants, the MS nutrient medium was supplemented with 3 mg/L of 6-benzylaminopurine (BAP), 2 mg/L of zeatin, 0.1, 0.5 and 1 mM of glyphosate, respectively. As a control, glyphosate-free regenerative nutrient medium was used. The results were statistically processed using Microsoft Excel. Results. It has been shown that glyphosate-susceptible rape explants form leaves and shoots with a frequency up to 8.3 % ± 0.8 % on the medium supplemented with 0.1 mM glyphosate compared to 57.2 % ± 0.6 % in the control. However, the morphogenic structures did not pass selection after transferring shoots to the elongation medium supplemented with 0.1 mM glyphosate. Conclusions. For the in vitro selection of rapeseed 0.1 mM glyphosate can be used in a nutrient medium at the stage of adventitious buds formation. But at the regeneration stage, the amount of herbicide should be increased to avoid false-positive results. Keywords: winter rapeseed, Brassica napus L., in vitro culture, selective marker, herbicide, glyphosate.

scholarly journals Screening Pyrus Germplasm for in Vitro Rooting Response

HortScience ◽  
1995 ◽  
Vol 30 (6) ◽  
pp. 1292-1294 ◽  
Author(s):  
Barbara M. Reed
Keyword(s):  
Acetic Acid ◽  
Growth Regulators ◽  
Additional Treatment ◽  
In Vitro Rooting ◽  
Root Production ◽  
Naphthaleneacetic Acid ◽  
Indole 3 Acetic Acid ◽  
Rooting Response ◽  
Selection Of

Micropropagated shoots of 49 Pyrus species and cultivars and one selection of Pyronia veitchii (Trabut) Guillaumin were evaluated to test their responses to several in vitro rooting techniques. Auxin treatment was required for rooting in most cases. Eighteen of 50 accessions rooted ≥50% with a 15-second, 10-mm IBA dip followed by growth on medium with no growth regulators (NGR). Twelve accessions rooted on a medium with 10 μm IBA applied for 1 week followed by NGR medium for 3 weeks; NGR medium alone was effective for only two accessions. Twenty-eight accessions rooted poorly with IBA treatments; an additional treatment of a 15-second dip in 10 mm NAA followed by NGR medium produced ≥50% rooting for eight genotypes. Root production increased for 10 of 19 especially recalcitrant genotypes by 10 μm IAA treatments in darkness or at 30C and NAA dip treatments. Of rooted shoots, 73% survived acclimation in the greenhouse. Selections of Pyrus betulifolia Bunge, P. calleryana Decne., P. hondoensis Kikuchi and Nakai, P. koehnei C. Schneider, P. pashia Buch.-Ham. ex D. Don, P. pyrifolia (Burm.f.) Nakai cv. Shinseiki, P. regelii Rheder, P. ussuriensis Maxim., and the Pyronia veitchii selection failed to root in any of the treatments. Twenty-five of 32 P. communis L. cultivars and three other species rooted on at least one of the treatments. Chemical names used: 1-naphthaleneacetic acid (NAA), 1H-indole-3-butyric acid (IBA), 1H-indole-3-acetic acid (IAA).

scholarly journals Does in vitro selection of biocontrol agents guarantee success in planta? A study case of wheat protection against Fusarium seedling blight by soil bacteria

PLoS ONE ◽  
2019 ◽  
Vol 14 (12) ◽  
pp. e0225655 ◽  
Author(s):  
Yoann Besset-Manzoni ◽  
Pierre Joly ◽  
Aline Brutel ◽  
Florence Gerin ◽  
Olivier Soudière ◽  
...  
Keyword(s):  
Soil Bacteria ◽  
In Vitro Selection ◽  
Biocontrol Agents ◽  
Seedling Blight ◽  
In Planta ◽  
Fusarium Seedling Blight ◽  
Study Case ◽  
Selection Of

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 endoplasmic reticulum is partitioned asymmetrically during mitosis before cell fate selection in proneuronal cells in the early Drosophila embryo

2017 ◽  
Vol 28 (11) ◽  
pp. 1530-1538 ◽  
Author(s):  
Anthony S. Eritano ◽  
Arturo Altamirano ◽  
Sarah Beyeler ◽  
Norma Gaytan ◽  
Mark Velasquez ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Division ◽  
Cell Fate ◽  
Asymmetric Cell Division ◽  
Asymmetric Division ◽  
Drosophila Embryo ◽  
Early Drosophila Embryo ◽  
Dividing Cells ◽  
Fate Determinants ◽  
Selection Of

Asymmetric cell division is the primary mechanism to generate cellular diversity, and it relies on the correct partitioning of cell fate determinants. However, the mechanism by which these determinants are delivered and positioned is poorly understood, and the upstream signal to initiate asymmetric cell division is unknown. Here we report that the endoplasmic reticulum (ER) is asymmetrically partitioned during mitosis in epithelial cells just before delamination and selection of a proneural cell fate in the early Drosophila embryo. At the start of gastrulation, the ER divides asymmetrically into a population of asynchronously dividing cells at the anterior end of the embryo. We found that this asymmetric division of the ER depends on the highly conserved ER membrane protein Jagunal (Jagn). RNA inhibition of jagn just before the start of gastrulation disrupts this asymmetric division of the ER. In addition, jagn-deficient embryos display defects in apical-basal spindle orientation in delaminated embryonic neuroblasts. Our results describe a model in which an organelle is partitioned asymmetrically in an otherwise symmetrically dividing cell population just upstream of cell fate determination and updates previous models of spindle-based selection of cell fate during mitosis.

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