scholarly journals Creation of the bank of breeding material of winter rye in vitro

2021 ◽  
Vol 1 (99) ◽  
pp. 29-37
Author(s):  
L. O. Ryabovol ◽  
◽  
Ya. S. Ryabovol ◽  
I. P. Diordiieva
Keyword(s):  
Light Intensity ◽  
Nutrient Medium ◽  
Plant Material ◽  
Storage Temperature ◽  
Selection Process ◽  
Winter Rye ◽  
Low Light ◽  
Breeding Material

Аn important issue of the selection process is the preservation of the source and obtained material, especially cross-pollinated crops, which significantly lose their viability by inbreeding. Creating a bank of genetic resources using biotechnological methods will effectively solve this problem. The aim of the work was to determine the conditions for the formation of a genetic bank of valuable winter rye materials with changes in temperature and modeling of the nutrient medium for long-term disposal of cloned plants and the use of active collection of original and created forms in the selection process. To deposit the clones, a nutrient medium, which included macro- and microelements according to the Murashige-Skuga medium was used. The nutrient substrate modified with cytokinins and carbohydrates. The clones in culture rooms at a temperature of 6–12 °С and low light intensity (2 kLk) were stored. In the course of research the conditions of creation of an active collection of plants of winter rye with use of temperature restriction and modification of a nutrient medium are defined. A consistent technological scheme for the conversion of plant material into a state of relative anabiosis has been developed. It is proved that the optimal storage temperature for samples is 6 °С. Survival of plants at the specified temperature regime after 12 months of deposition on average by genotypes at the level of 78,2 % was recorded. Modification of the nutrient medium with agar-agar at a concentration of 12,0 g/l increases the proportion of viable clones to 81,3 %, and the introduction into the substrate of an increased concentration of growth regulators, in particular 6-BAP (2,0 mg/l) and sucrose 40,0 g/l and a gradual decrease in temperature to 10 °С prolongs the period of deposition of cloned plants without changing the substrate and the shelf life of breeding material in isolated crops. Using of biotechnological methods for the preservation and reproduction of valuable material intensifies the selection process of obtaining initial samples of winter rye.

scholarly journals PHYSIOLOGICAL ASPECTS OF IN VITRO-GROWN COCONUT (Cocos nucifera L.) PLANTS DURING ACCLIMATIZATION

CORD ◽  
1999 ◽  
Vol 15 (02) ◽  
pp. 34
Author(s):  
C S Ranasinghe ◽  
L K Weerakoon ◽  
Y M H Liyanage ◽  
D T Mathes
Keyword(s):  
Light Intensity ◽  
Chlorophyll Content ◽  
Environmental Conditions ◽  
Cocos Nucifera ◽  
Low Light ◽  
Leaf Chlorophyll Content ◽  
Leaf Chlorophyll ◽  
Low Light Intensity ◽  
Cocos Nucifera L

The physiological status of in vitro-grown coconut (Cocos nucifera L.) plants during acclimatization was studied using nursery-raised seedlings as the control.  The percentage of open stomata in leaves of in vitro-grown coconut plants was high at the initial stage of acclimatization but decreased during the course of acclimatization indicating an improvement in stomatal regulation.  A progressive increase in the stomatal density, epicuticular wax deposition and leaf thickness in in vitro-grown plants was observed during acclimatization. As a result of the low light intensity, the epidermal cells of in vitro-grown plants were narrower and longer when compared to the control.  With the exposure of plants to increased light intensity, the cells became wider and shorter as observed in the control.   The leaf chlorophyll content was high in in vitro-grown plants under low light intensity.  With increasing light intensity, a reduction in leaf chlorophyll content in vitro-grown plants was observed and at the later stages of acclimatization, it was comparable to that of the control. Variations in the rates of photosynthesis and transpiration in vitro-grown plants were observed in response to the changing environmental conditions.  However, at the end of acclimatization, where the plants were ready to be transferred to the field, the physiological statuses of in vitro-grown coconut plants were comparable to that of nursery raised seedlings. The present study revealed that the embryo-cultured coconut plants could adjust well to the changing environmental conditions during acclimatization.

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 Storage and Viability Testing of Protea Pollen

1996 ◽  
Vol 121 (5) ◽  
pp. 804-809 ◽  
Author(s):  
I. David van der Walt ◽  
Gail M. Littlejohn
Keyword(s):  
Storage Temperature ◽  
Pollen Viability ◽  
Germination Percentage ◽  
Long Term Storage ◽  
Fresh Pollen ◽  
The Relationship ◽  
Term Storage ◽  
Storage Temperatures

The influence of storage temperature and humidity on pollen viability was studied in four Protea species. Pollen was stored at a range of temperatures and relative humidities for up to 1 year and tested for ability to germinate in vitro. Pollen of P. repens (L.) L. `Sneyd', P. eximia (Salisb. ex Knight) Fourcade `Fiery Duchess' and P. magnifica Link. clone T 84 07 05 stored at -196 °C and -14 to -18 °C retained a germination percentage as high as that of fresh pollen regardless of humidity. Humidity control became increasingly important at storage temperatures above 0 °C. The study showed that long-term storage of Protea pollen is not feasible at temperatures above 0 °C. The relationship between germinability and fluorochromasia (FCR) was studied during storage of `Sneyd' pollen. The correlations between FCR and germinability were found to be low and nonsignificant. Fifteen-month-old cryopreserved `Sneyd' pollen functioned in fertilization and seed set as effectively as fresh pollen.

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 RADIATION FOR GARLIC PLANLETS ON ACLIMATIZATION PHASE

2015 ◽  
Vol 30 (1) ◽  
pp. 25
Author(s):  
Dara Cipta Andini ◽  
Eddy Tri Haryanto ◽  
Djoko Purnomo
Keyword(s):  
Virus Infection ◽  
Light Intensity ◽  
Plant Material ◽  
In Vitro Propagation ◽  
Seed Plant ◽  
Horticultural Crops ◽  
Long Time ◽  
The Impact

<p><em>Garlic is one of the important horticultural crops in Indonesia so that the needs increas from year to year. In Indonesia plant material (seed) for cultivating garlic always uses comsumption bulb. The impact of that technique in a long time that the yield gradually decrease by virus infection from preliminary crop. The aim of the research was to find out of seed plant material which virus free by in vitro propagation technology. Acclimatitation is one of succesfuly determination in invitro propagation. Research by the experiment was to study the ability of New Tawangmangu, Gunung Kidul, and Bali garlic varieties on acclimatitation in several light intensity (by paranet shading). The results showed that the totipotency of the garlic varieties was high but in acclimatitation by the shading technique just survived two weeks only. The failure of acclimatitation was range of temperature 12-37</em><em>°C</em><em>.</em></p><p> </p>

scholarly journals Recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. as a source for synthetic seed production for medium-term preservation

2012 ◽  
Vol 64 (2) ◽  
pp. 809-817 ◽  
Author(s):  
Irina Holobiuc ◽  
R. Catana
Keyword(s):  
Somatic Embryogenesis ◽  
Seed Production ◽  
Somatic Embryos ◽  
Plant Material ◽  
Synthetic Seed ◽  
Sugar Alcohols ◽  
Medium Term ◽  
Gentiana Lutea

Our aim was to establish an efficient and reproducible system for producing synthetic seeds from recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. This species is a vulnerable medicinal plant, protected both at the national and international levels, and is included in different Red Lists and Books. In vitro culture, as an alternative to classical methods of preservation, allows for the cyclic multiplication of plant material and short-, medium- and long-term preservation of tissue collections. Biotechnological approaches allow for maintenance of the plant material in a confined space and protection against biotic and abiotic factors. Somatic embryogenesis (SE) is the most efficient way to regenerate plants, ensuring material for preservation and fundamental research. In our experiment, recurrent somatic embryogenesis was developed in long-term cultures in the presence of sugar alcohols (mannitol, sorbitol) and in the absence of growth factors. This process proceeded at a high rate, with adventive somatic embryos being generated in a continuous process, followed by maturation, germination and development into plants. To follow the somatic embryogenesis process, histological samples were made. We used these embryogenic cultures for synthetic seed production and medium-term conservation. The viability of somatic embryos after moderate osmotic stress treatment was tested using TTC. Our methodology relied on the induction of somatic embryogenesis in the presence of auxins in the first cycle of in vitro cultures, long-term high embryogenic culture maintenance in the presence of sugar alcohols and synthetic seed production.

Expression analysis of five candidate genes in eight contrasting rice genotypes suggest role of Lagging growth and development 1(LGD1), Pinhead1 (PNH1) and Leaf Panicle 2 (LP2) in low light intensity response at vegetative stage

2020 ◽  
Vol 57 (4) ◽  
pp. 261-270
Author(s):  
Suvendhu S Dutta ◽  
Wricha Tyagi ◽  
Mayank Rai
Keyword(s):  
Light Intensity ◽  
Candidate Genes ◽  
Light Stress ◽  
Vegetative Stage ◽  
Low Light ◽  
Expression Levels ◽  
Low Light Intensity ◽  
Rice Genotypes ◽  
Low Light Stress

Light acts as an energy source in plants for photosynthesis and also is an environmental cue leading to growth and differentiation. The quality and duration of light is therefore, key to obtaining yield and growth targets. Our previous study in rice led to identification of a panel of contrasting genotypes and putative candidate genes and markers for low light intensity tolerance. In the present study, expression of a set of five candidate genes (LGD1, PNH1, ILA1, CAB2R and LP2) previously identified to be associated with low light intensity tolerance was studied in a panel of eight rice genotypes at two time points post stress induction (one hour and two days). Cumulative normalised expression levels for genes LDG1 and PNH1 were down-regulated in response to one hour low light stress only in susceptible genotypes. While the cumulative normalised expression levels of ILA1 and LP2 genes were down- and up-regulated, respectively in tolerant genotypes. After two days of low light stress, expression of PNH1 and LP2 were down- and up-regulated in tolerant and susceptible genotypes, respectively. Our data suggests that genes LGD1, PNH1 and LP2 can be targeted for achieving vegetative stage low light intensity tolerance. Long term stress followed by recovery experiment revealed genotype Pusa Sugangh-5 as tolerant to low light intensity. These experiments suggest that mechanism of low light intensity tolerance in Pusa Sugangh-5 is distinct from the other four tolerant rice genotypes.

scholarly journals Induction of chlorophyll fluorescence use for assessment of functioning of Gentiana lutea L. plants photosynthetic apparatus in different conditions of culturing in vitro

2019 ◽  
Vol 25 ◽  
pp. 209-214
Author(s):  
L. R. Hrytsak ◽  
A. I. Herts ◽  
N. V. Herts ◽  
N. M. Drobyk
Keyword(s):  
Chlorophyll Fluorescence ◽  
Light Intensity ◽  
Nutrient Medium ◽  
Photosynthetic Apparatus ◽  
Resistance Mechanisms ◽  
Wave Band ◽  
Light Conditions ◽  
Pam Fluorometry ◽  
Gentiana Lutea

Aim. To study the peculiarities of functioning of photosynthetic apparatus of Gentiana lutea L. plants in vitro under different light qualities and source of carbon in the composition of nutrient medium by using the induction of chlorophyll fluorescence (ICF) method. Methods. Chlorophyll fluorescence was determined in light-adapted leaves of cultivated in vitro G. lutea plants by use of PAM fluorometry MultispeQ. The parameters change of photosynthetic apparatus functioning of cultivated in vitro plants was assessed with regard to light qualities (Variant 1: light intensity – 85 W/m2, when different spectra combinations of blue-wave band (Еb) and green-wave band (Еg) and red-wave band (Еr) was 33% : 42% : 25%; Variant 2: light intensity – 100 W/m2, with wave bands of spectra – Еb : Еg : Еr = 25% : 27% : 48%) and the source of carbon (10 g/l of sucrose or 3 g/l of mannite) in the composition of nutrient medium МS/2 (MS medium with half amount of macro- and microsalts), supplemented with 0.1mg/l of kinetin. Results. It was established that G. lutea plants cultivated in vitro for 90 days in light conditions of Variant 2 had a quantum yield photochemical of PS ІІ 8.3 % higher in comparison to G. lutea specimens cultured in light conditions of Variant 1. The vitality value of Variant 2 plants was 23 % higher. Change of carbon source in the composition of nutrient medium from sucrose (10 g/l) to mannite (3 g/l) was both increasing the efficiency of functioning of photosynthetic apparatus of Gentiana lutea L. plants in vitro and activating their resistance mechanisms to water deficit. Conclusions. The ICF method use shows that functioning of photosynthetic apparatus of G. lutea plants in vitro depends different qualities light and source of carbon in the composition of nutrient medium. Key words: induction of chlorophyll a fluorescence, plants in vitro, Gentiana lutea L.

scholarly journals Can protoplast production from in vitro cultured shoots of Tanacetum vary during the season?

2001 ◽  
Vol 10 (3) ◽  
pp. 145-151 ◽  
Author(s):  
M. KESKITALO
Keyword(s):  
Light Intensity ◽  
High Light Intensity ◽  
High Light ◽  
Shoot Tips ◽  
Low Light ◽  
Tanacetum Vulgare ◽  
Chemical Changes ◽  
Tanacetum Cinerariifolium ◽  
Protoplast Production

Two different experiments were carried out to study the production of protoplasts and the variation of protoplast yield from in vitro cultured shoot tips of tansy (Tanacetum vulgare L.) and pyrethrum (Tanacetum cinerariifolium (Trevir.) Schiltz-Bip). In the first experiment, light had more pronouced effect for tansy than for pyrethrum. When the donor tissues of tansy were cultured under high light intensity the leaves contained anthocyanin and became brown during enzyme maceration. In contrast, donor tissues cultured under low light intensity produced leaves without anthocyanin. Depending on the light intensity of donor tissues, on average 5.8 - 6.8 x 106 and 3.4 - 4.3 x 106 protoplasts were isolated from one gram of mesophyll leaves of tansy and pyrethrum, respectively. In the second experiment, the production of protoplasts from tansy and pyrethrum varied seasonally. The most successful season for the production of protoplasts from in vitro cultured shoot tips was between December and April, when also the highest number of protoplasts could be isolated. It was not possible to state whether Tanacetum species have rhythms, which could cause physiological or chemical changes for the in vitro grown shoot tips. However, some external or internal, possible seasonal-dependent stimuli may have caused variation in the number of protoplasts isolated from tansy and pyrethrum and favoured protoplast production during winter and spring.

scholarly journals Media Composition and Light Affect Storability and Poststorage Recovery of Micropropagated Hosta Plantlets

HortScience ◽  
2000 ◽  
Vol 35 (6) ◽  
pp. 1159-1162 ◽  
Author(s):  
Sandra B. Wilson ◽  
Nihal C. Rajapakse ◽  
Roy E. Young
Keyword(s):  
Storage Temperature ◽  
Visual Quality ◽  
Dry Weight ◽  
Poor Quality ◽  
Photon Flux ◽  
Storage Duration ◽  
Long Term Storage ◽  
Leaf Yellowing

Hosta (Hosta tokudama Makeawa `Newberry Gold') plantlets were micropropagated photoautotrophically (without sucrose in medium) or photomixotrophically (with 2% sucrose in medium) for 3 weeks at 23 °C under 80 μmol·m-2·s-1 photosynthetic photon flux (PPF) prior to long-term storage. Plantlets were stored for 4, 8, or 12 weeks at 5, 10, or 22 °C in darkness or under white (400-800 nm), blue (400-500 nm), or red (600-700 nm) light at or near light compensation points. Illumination during storage was necessary to maintain dry weight and regrowth potentials of plantlets in vitro, but light quality had no effect on these parameters. All photoautotrophic plantlets stored in darkness were of poor quality at the time of removal from storage and died when transferred to the greenhouse. Dark-stored photomixotrophic plantlets survived storage for 12 weeks at 5 °C, but declined in appearance (visual quality) as the storage duration increased. Decline in visual quality was greater when plantlets were stored at 10 and 22 °C. Leaf dry weight of illuminated plantlets increased and percentage of leaf yellowing decreased as storage temperature increased. Recovery of illuminated plantlets from photomixotrophic storage was best when plantlets were stored at 22 °C. These plantlets were characterized by increased visual quality (color and form) and increased dry weight compared with those in other treatments. After 60 days in the greenhouse, the dry weight of these plantlets was similar for 4-, 8-, and 12-week storage durations, indicating flexibility in storage time if specific light and temperature provisions are met.

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