MS Nutrient Medium Research Articles

Effect Of Cytokinin BA And Different Concentrations Of Chemical Stress Treatments On The Multiplication Of Vegetative Shoots Of The Plant Antirrhinum Magus L. Grown In Vitro

The study was conducted on the multiplication of Antirrhinum magus L. plant using the micropropagation technique, represented by culture vegetative shoots on MS nutrient medium prepared with different concentrations of cytokinin BA and different levels of chemical stress treatments. The treatment of 4.44 μmol.L–1 BA with a NAA concentration of 0.54 μmol.L–1 gave the highest significant increase in the average number of vegetative shoots per plantlet, shoot length, number of leaves per plantlet, and leaf width, which amounted to 6.40 shoot explant-1, 4.00 cm, 20.60 shoot leaves -1, 0.68 cm, respectively, while a significant decrease in the average vegetative traits was observed when the concentration of BA in the medium was increased in the two treatments (8.88, 11.10) μmol.L–1 BA, which did not differ significantly between them. The results of chemical stress showed that reducing the level Mannitol to 15gm L-1 in multiplication medium led to a significant and inverse increase in the average number of vegetative shoots per plantlet, shoot length, number of leaves per shoot, and leaf width reached 4.80 shoot explant-1, 3.50 cm, 11.40 shoot explant1-, 0.44 cm in succession, and their averages decreased. By increasing the concentration in the middle. It was also observed that there was an inverse relationship between the average vegetative traits and the levels of PEG8000, where the control treatment achieved the highest significant increase in the average of all vegetative traits, amounting to 6.60 shoot explant-1, 4.06 cm, and 7.60 shoot explant-1, 0.92 cm, respectively, compared to the other averages of water stress whenever the level of PEG8000 increased. In the nutrient medium down to the level of 45g. L-1.

OPTIMIZATION OF THE HORMONAL STATUS OF THE CULTURE MEDIUM FOR H. RHAMNOIDES IN VITRO CULTIVATION

Sea buckthorn (Hippophae rhamnoides L.) is an ancient crop that has traditionally been used as a medicinal plant, health promotion agent and dietary supplement. The fruits of sea buckthorn, which are a source of carotenoids, which had been collected since ancient times and was introduced into cultivation only at the beginning of the 20th century. Currently, sea buckthorn berries are harvested on an industrial scale, as due to the high content of biologically active substances in them, consumer interest in sea buckthorn products has increased significantly, and their use in the food industry and medicine is expanding. Due to its biologically active compounds, high content of vitamin C, flavonoids, carotenoids and tocopherols, sea buckthorn is an excellent raw material for functional foods. The purpose of this work is to select conditions for cultivating explants of H. rhamnoides L. plants and obtaining micro-shoots for subsequent microclonal reproduction. The material for initiating a sterile culture was the seeds H. rhamnoides. The samples were introduced from natural populations in the RSE “Altai Botanical Garden” of the SC of MoES of the Republic of Kazakhstan (Ridder). Solutions of 0.1% sulema, 34% hydrogen peroxide, and 10% Domestos solution were used for seed sterilization, the exposure period was 20 minutes. Next, the seeds were planted on the surface of the WPM and ½ MS nutrient medium with the addition of phytohormones 6-benzylaminopurine (BA) and gibberellic acid (GA3) in various concentrations. Activated carbon in an amount of 2 g/l was used to reduce the accumulation of polyphenols in the medium. The beginning of germination was noted on days 7-8, depending on the genotype of sea buckthorn. The minimum number of infected seeds was observed in the variant using 0.1% sulema. When using hydrogen peroxide, the contamination of seeds with fungal and bacterial infections was at the level of 40.8-45.5%. The largest number of germinated seeds were observed when using WPM medium. The maximum values of seedling length were noted on WPM media with the addition of 1 mg/l BA + 1 mg/l GA3, where the average shoot length was 8.6±0.9 cm. In this variant, sterile seedlings were characterized by an intense green color and well-developed cotyledon leaves. In some genotypes, intensive growth and the appearance of a second internode were noted. In other variants, height indicators varied from 3.2 cm. up to 4.2 cm. Thus, we have found that WPM medium supplemented with 1 mg/l 6-benzylaminopurine and 1 mg/l gibberellic acid is better suited for obtaining sterile shoots from H. rhamnoides seeds. The obtained sterile seedlings of H. rhamnoides will be used in vitro culture for subsequent microclonal reproduction.

Use of the Sulfur-Containing Growth Regulator Tiaton for Rooting Micro-Gears of Grapes in Culture in vitro

The study was carried out to evaluate the effect of a nutrient solution for the rhizogenesis of grape micro–gears in the Murasiga-Skuga medium (MS) with the addition of various concentrations of the drug Tiaton (0.1, 0.15, 0.20 and 0.25 ml/l). In experiments, the effect of different concentrations of the drug Tiaton on the rooting and vegetative growth of grapes of the Khasansky and Moskovsky White hybrids in vitro was studied. When the Tiaton preparation was added to the MS nutrient medium in various concentrations, the rooting capacity of the micro-gears of the Khasansky hybrid grape increased by 40%, the total root length increased by 2.4 times compared with the control. In the Moscow White hybrid, when using the Triaton growth regulator, an increase in the rooting capacity of micro gears up to 25% and the total root length by 1.5 times is shown. The optimal concentrations of the Triaton preparation for addition to the MS nutrient solution for optimal rhizogenesis of micro-gears and the development of vegetative growth of cuttings of the studied grape hybrids have been established. Thus, for Khasansky hybrid grapes, the most optimal medium for improving rooting and root development of micro gears was ½ MS + 0.15 ml/l of Tiaton preparation. In the Moscow White hybrid, a significant increase in the rooting capacity of micro-gears and good development of the root system was obtained when Tiaton was added to the nutrient medium for rhizogenase of micro-gears in concentrations (0.15 and 0.20 ml/l).

The influence of the nutrient composition medium on the intensity micropropagation in vitro Fragaria × Ananassa duch.

Strawberry is one of the most economically significant crops in the world berry growing. In the course of research, the influence of the mineral composition of nutrient media on the growth and development of strawberry plants at the stage of micro-propagation proper was studied for further optimization of the technology of microclonal reproduction. The purpose of the work is to select the optimal nutrient medium for micropropagation of strawberry in vitro in the production system of healthy planting material. The research was carried out in the laboratory of biotechnology of the Russian research Institute of Fruit Grop Breeding according to generally accepted methods. The objects were important commercial varieties of garden strawberries (Fragaria × ananassa Duch.) of foreign breeding: Alba (NF 311), Darselect, Kimberly, Clery, Syria (NF 137), Florence. Variants of the nutrient media formulations used: Lee and de Fossard, Gamborg and Eveleigh (B5), Knop, Murashige-Skoog with the addition of 0.8 mg/ l cytokinin 6-BAP. Optimal recipes of the nutrient medium that promote intensive proliferation and root formation in plants have been determined. For the cultivation of the studied strawberry varieties at the stage of actual micro-propagation with a high degree of regeneration, the optimal nutrient medium is the MS medium. On the nutrient Knop’s medium, the micro-plants on the second passage acquired a reddish color, not characteristic of the studied varieties. The maximum values of plant height were noted in the control variant with MS nutrient medium in the varieties Darselect (9.1 mm) and Kimberly (8.6 mm). In all other variants of the studied nutrient media, the average height of plants did not exceed 7.7 mm. Media according to the LF and B5 recipe can be used to obtain microplants suitable for planting in ex vitro conditions, excluding the rooting stage, to accelerate the production of healthy planting material. A different reaction of strawberry varieties to the mineral composition of nutrient media has been established.

Anatomy, ploidy level, and essential oil composition of <i>Hyssopus officinalis</i> ʻNikitskiy Beliyʼ <i>in vitro</i> and <i>ex situ</i>

Background. Clonal micropropagation is a biotechnological method for plant multiplication. The existing data on the structure of organs in vitro, genetic stability, and essential oil composition are limited for Hyssopus officinalis L., so this study was aimead at investigating these aspects under a short period of in vitro culturing.Materials and methods. Plants of Hyssopus officinalis ʻNikitskiy Beliyʼ cultivated ex situ, in vitro and ex vitro were analyzed. Conventional methods were applied to study plant anatomy, ploidy level, and relative DNA content, as well as to extract and analyze essential oil. Statistical analysis was performed using the Past 4.03 software.Results. According to the results obtained, with 6-BAP introduced into MS nutrient medium in optimal concentrations (0.3– 0.5 mg/L), the general in vitro structure of leaf blades in the developed microshoots was similar to those in ex situ plants, while the qualitative and quantitative changes observed were induced by the effect of specific culturing conditions and plant rejuvenation. The analysis of the ploidy level and relative DNA content in the nuclei isolated from the leaf tissue cells of the microshoots ex vitro after adaptation revealed no changes compared to the ex situ leaf parameters. The mass fraction of essential oil and its component composition in the mother plants and ex vitro regenerants were similar.Conclusion. Cultivation of Hyssopus officinalis ʻNikitskiy Beliyʼ microshoots on MS nutrient medium with 6-BAP optimal concentrations promotes morphogenesis without significant deviations in the ploidy level, relative DNA content, essential oil yield, or its component composition. The developed protocol for clonal micropropagation of Hyssopus officinalis ʻNikitskiy Beliyʼ provides clones identical to the ex situ plants.

Somaclonal Variation in Callus Cultures of Rose Periwinkle, <i>Catharanthus Roseus L</i>. Under Induced Salt and Osmotic Stresses

The experiment was carried out at the laboratories of the National Commission for Biotechnology in Syria during the period 2020-2022 to detect the somaclonal genetic variation occurring in callus cultures at different ages and to compare them with the <em>In vitro</em> growing plants<em> </em>and<em> </em>introduced species of Rose Periwinkle that grew up in the Syrian environment, in addition, exploring the effect of abiotic stresses in causing genetic variations in callus, by the Inter Simple Sequence Repeats technique (ISSR) using 21 primers. Seeds were germinated on an MS nutrient medium devoid of growth regulators; explants were transferred to a propagation medium fortified with Naphthalene Acetic Acid (NAA) (1 mg L<sup>-1</sup>) and 6-Benzyladenine (BA) (2 mg L<sup>-1</sup>). Callus was developed from Rose periwinkle <em>Catharanthus roseus L</em>. leaves using 5C01 medium fortified with NAA and Kinetin (Kin). After that, callus was exposed to gradually increasing levels of PEG-6000-induced osmotic stress and salt stress (NaCl) with a higher level every 30 days. The molecular study results proved the effectiveness of all the used primers (21 primers) in presenting polymorphism between the studied samples at a rate of 92.16%. The cluster analysis showed separating the analyzed samples into two main clusters. The first cluster contained the osmotic stressed callus, while the second cluster separated into two sub-clusters that had the rest of the studied samples. It was found that there were genetic variations between <em>In vitro</em> plants and the callus. In addition, the stressed callus (salt and osmotic) was compared to the unstressed callus, and this confirmed that callus cultures were an essential source of genetic variations, and the ISSR technique was an effective tool for detecting these variations.

Peculiarities of plant adaptation of interspecific hybrid Betula ex vitro

Microclonal propagation as one of the methods of biotechnology allows obtaining genetically homogeneous plants during the year from a minimum amount of donor material. Adaptation of plants ex vitro to environmental conditions is the final and important stage of microclonal propagation. The purpose of the study is to determine the optimal mode of plant adaptation of triploid interspecific hybrid Betula ex vitro to environmental conditions. Biotechnological (microclonal propagation) and statistical (arithmetic mean, standard error, one-way analysis of variance (ANOVA)) methods were used for research. The study was carried out in the research of Plant Biotechnology Laboratory of the Separated Subdivision of National University of Life and Environmental Sciences of Ukraine “Boyarka Forest Research Station” during 2019-2023. According to research, the MS nutrient medium with the addition of 0.25 mg∙l -1 of kinetin stimulated the active proliferation of microshoots and root system. Optimum plant development took place on the nutrient medium vitro for a 25-30- day cycle of cultivation, which contributed to successful adaptation to environmental conditions. The proportion of adapted Betula ex vitro hybrid plants on agroperlite substrate was more than 60% for 30 days. Significant plant viability (more than 80%) was obtained under the conditions of preliminary exposure of the plant root system in auxin solution (1.0 mg/lIAA, 1.0 mg/l NAA, 1.0 mg/lIBA) for 25-30 minutes followed by daily spraying of leaves with 20% glycerin for 15-20 days. Adapted plants had typical pigmentation without signs of chlorosis and vitrification. In the spring, the survival rate of plants under environmental conditions was more than 80% and indicated a high level of adaptability after in vitro. The developed protocol for the adaptation of the Betula ex vitro hybrid allows obtaining high plant survival in environmental conditions. The practical value of the research is obtaining plants adapted to environmental conditions ex vitro, which in the future can be used to qualitatively enrich the range of artificial plantings in settlements

Optimizing Different Medium Component Concentration and Temperature Stress Pretreatment for Gynogenesis Induction in Unpollinated Ovule Culture of Sugar Beet (Beta vulgaris L.)

The great economic importance of sugar beet determines the ongoing biotechnological studies conducted worldwide to improve the technology of obtaining doubled haploids (DHs) using the method of unpollinated ovule culture in vitro. To improve the induction of gynogenesis, we tested the effect of thidiazuron (TDZ), temperature bud pretreatment, different concentrations of sucrose, and culturing on liquid or solid medium. Three genotypes were tested in this study. The use of TDZ at a concentration of 0.4 mg/L in solid IMB (induction medium for Beta vulgaris) induction nutrient medium with 3 g/L phytagel, 50 g/L sucrose, 200 mg/L ampicillin and cultivation at 28◦C in the dark produced up to 16.7% induced ovules. The liquid nutrient medium of the same composition induced up to 8% ovules. Increasing TDZ concentration to 0.8 mg/L resulted in reduction or total inhibition of gynogenesis, depending on the genotype. Reducing the sucrose concentration to 20 g/L or increasing it to 80 g/L was not effective. In all three genotypes, the absence of temperature pretreatment of buds (5–6 °C) showed the best results. The plant regeneration with MS nutrient medium of 20 g/L sucrose, 3 g/L phytagel, 1 mg/L 6-benzylaminopurine (BAP) and 0.1 mg/L gibberellic acid (GA3) resulted in up to seven shoots from one induced ovule in the most responsive genotype. We showed by flow cytometry, chromosome counting and chloroplast number assessment that all regenerant plants were haploid (2n = x = 9).

Determination of effective surface sterilization protocol in in vitro tissue culture for Giant Snowdrop (Galanthus elwesii Hook) bulbs

Giant Snowdrop (Galanthus elwesii Hook) is a species of snowdrop belonging to the Amaryllidaceae family. In this study, the deformation rates of the bulbs was calculated as % (percent). For sterilization G. elwesii bulbs were sterilized at different concentrations (1, 3, 5, 7, 9, 11 and 13 %) of sodium hypochlorite (NaOCl) for 5, 10, 15, 20, 25 minutes. The bulbs were rinsed with sterile distilled water 3 times for 5 minutes and then transferred to the MS medium. Contamination rates in MS nutrient medium were determined as percent (%) after 7 days. Deformation was not observed in the bulbs sterilized with 1 and 3% sodium hypochlorite solution however 100% contamination was detected. The most effective surface sterilization was obtained by soaking in 9-11% sodium hypochlorite solution for 5 and 10 minutes. As the sodium hypochlorite concentration and the application time increased, the surface sterilization of the bulbs increased, but the deformation rate of the scaly leaves of the bulbs increased due to this increase.

Functional microbiome of Ananas spp. as growth-promoting agent in pineapple BRS Imperial

The pineapple cv. BRS Imperial developed by Embrapa Mandioca e Fruticultura has excellent agronomical and sensorial attributes with great market acceptance. However, the cultivation of plants is time-consuming with low performance and slow growth, limiting adoption especially by large-scale pineapple growers. Therefore, the aim of this study was to evaluate the growth potential of bacteria from the associated microbiome of Ananas spp. in order to validate its application in different stages of cultivation (semi-field, in vitro and greenhouse), for the production of seedlings on a large scale. Eighteen treatments consisting each of a different bacteria from the biological collection of microorganisms associated with the Ananas genus, were tested under semi-field conditions. Three treatments (BAC25, BAC222 and BAC406) were selected to validate the microbiolization process of pineapple seedlings from the cv. BRS Imperial in vitro (rooting stage) and in vivo (acclimatization in the greenhouse) together with a mixed treatment (composed by the combination of the three bacteria) and a control treatment (untreated plants). The in vitro cultivation was carried out in commercial substrate autoclaved with MS nutrient medium where bacterial suspensions were added according to the treatments mentioned above. Each treatment consisted of 120 replicates, totaling 600 experimental units. The evaluations were carried out at weekly intervals for a period of 42 days, when destructive analyses were performed. The remaining plants were transferred to the greenhouse and evaluated from this stage until complete acclimatization. For the in vivo validation, the bacterial suspensions were applied to the substrates contained in plastic cups, with pineapple seedlings of the cv. 'BRS Imperial', totaling 200 plants per treatment. The evaluations consisted of measuring growth variables and destructive analysis. The data obtained were subjected to analysis of variance and the means grouped by the Scott-Knott test at 5% probability. The BAC25, BAC222 and BAC406 treatments promoted plant growth and biomass increment in semi-field conditions. BAC406 was the most promising in promoting growth of pineapple seedlings in vitro, while BAC222 was the best treatment for the in vivo cultivation. The results obtained demonstrate that the bacteria tested have great potential for use in the different stages of cultivation of micropropagated seedlings, improving growth, health and vigor of plants.

Propagation of Chia Plants Using Plant Tissue Culture Technique

Back Ground: The plant tissue culture technique is one of the biotechnologies that meansisolating a plant cell, tissue or organ, sterilizing it and cultivating it on sterile artificial foodmedia under conditions completely free of pathogens, and then the transplanted part developsinto a complete plant similar to the original from which it was taken. Under controlledenvironmental conditions of temperature, humidity and lighting, as defined by others as sterilecultivation of cells, tissues, organs and their components under certain chemical and physicalconditions in vitro.Objectives: This study aimed to propagate the medicinal chia plant using the technique ofplant tissue culture.Materials and Methods: The research was carried out in the Plant Tissue Culture Laboratoryof the College of Agriculture, Anbar University. Sterilized chia seeds were sown on the MSnutrient medium equipped with the GA3 growth regulator for the purpose of obtainingvegetative growths in the cultivation tubes, at the rate of one seed per vial, using tweezerspreviously sterilized with alcohol and flame. After obtaining the required growths, the stemnodes were cut into pieces 1 cm long and planted on MS medium prepared with differentconcentrations of BA (0.5, 1, 2, 3) mg L -1 . And by interfering with NAA (0.1, 0.2, 0.3) mg L -1with the aim of the emergence and multiplication of vegetative branches within a factorialexperiment with two factors (4 × 4) with 10 replications for each treatment. 16 hours and 8hours dark. In the rooting experiment, different concentrations of IBA (0, 1, 2) mg L -1 wereused, interfering with BR (0, 0.1, 0.2) mg L -1 , and measurements were taken after one month.
 Results: It was observed through the obtained results that there were significant differencesbetween the different concentrations of growth regulators used in the experiment in themultiplication and rooting of the branches.Conclusions: Regarding the doubling stage, there is an increase in the number of branches asthe concentration of the growth regulator BA increases. For rooting, the growth regulatorbrassinosteroids outperformed auxin indole butyric acid in root lengths and numbers.Keywords: Chia Plants, Gibberellin hormone, Leaves, Stem Nodes, Tissue Culture Technique

Effect of Medium Components on in vitro Shoot Formation and Rooting of Papaya (Carica papaya L.)

The effect of modification of some selected MS nutrientmedium on in vitro growth and development of papaya (Carica papayaL.) shoot tips was examined with the objective of optimizing shootregeneration and rooting efficiencies. Modifications were made onselected components of MS medium. Shoot tip explants were obtainedfrom in vitro-grown plantlets. The results showed that MS salts at thetwice full strength was optimal for all parameters measured comparedwith other strengths tested. Sucrose concentrations of 0.75% and 1.5%were better than the other concentrations tested for growth anddevelopment of cultured shoot tips, and there was no significantdifference between them. The results revealed the importance of inclusionof BA and NAA in the nutrient medium at relatively low concentrations,the best results for shoot and leaf formation was recorded with thecombination of BA at 1.0 mg /litre and NAA at 0.01mg /litre, where thegreatest number of shoots and leaves was obtained. The combinationtreatment of 0.3 mg/litre BA and 0.00 mg/litre NAA recorded the bestresults for mean shoot elongation, compared with the other combinationtreatments tested. However, the produced roots were morphologicallyabnormal. The study attested to the essentiality of IBA for rooting of invitro produced papaya shoots, and the data suggested the conduction ofadditional studies for determination of optimal concentration for rooting

Creation of initial material using embryoculture and remote hybridization in the breeding of winter bread wheat

Aim. The research was to improve the technology of creating genetic diversity of samples obtained by remote hybridization of soft winter wheat and speltum wheat with usuing the biotechnological link in the selection scheme. Methods. Remote hybridization of plants is an effective method of obtaining new starting material. The research was conducted in the biotechnology laboratory of the Uman National University of Horticulture during 2018–2022. The initial maternal form was the soft winter wheat varieties Artemisia, Smuglyanka, Zolotokolosa and the parental form was the winter spelt variety Zorya Ukrainy. After hybridization, immature twelve-day-old embryos were planted on modified versions of MS nutrient medium and cultivated at 25 °C in the dark until macrostructures were formed. Results. The technology of obtaining hybrid material by remote hybridization of Triticum aestivum L. / Triticum spelta L. and its use in the selection scheme of the biotechnological link has been improved. Conclusions. A nutrient medium for the development of immature hybrid wheat germs has been developed. The highest yield of seedlings (40.0% on average by genotype) on the modified Murashige-Skug medium, which included 1,5 mg/l 6-benzylaminopurine, 0,5 mg/l indolylacetic acid, 1,0 mg/l gibberellin, 2,0 mg/l glutamine, 2,0 mg/l glycine, 0,5 mg/l serine, 100,0 mg/l meso-inositol, 50,0 g/l sucrose was recorded. It was noted that the induction of the development of biomaterial from hybrid immature embryos is a genetically determined factor. The possibility of obtaining genetic diversity of forms that can be used by donors of genes of individual traits using embryo culture was shown.

THE INFLUENCE OF ENDOGENOUS POLYPHENOLS, PHOTOPERIOD AND MINERAL COMPOSITION OF THE NUTRIENT MEDIUM ON THE FORMATION OF CALLUS TISSUE OF RELIC GYMNOSPERMS SEQUOIA SEMPERVIRENS (D.DON) ENDL.

Relrvance. Sequoia sempervirens (D.Don) Endl. are the tallest relict long–lived plants characterized by a limited range of growth. Valuable sequoia wood is able to accumulate unique secondary metabolites that do not have synthetic analogues. Sequoia plants grow in specialized microecological niches that limit their natural distribution area. It is known that biotic stress is one of the main environmental factors limiting the introduction of valua-ble plant species – possible sources of production of unique metabolites. Limiting environmental factors have a direct impact on the growth, develop-ment and productivity of secondary plant metabolism. Stress-resistant and highly productive plants can be created using cell biotechnology methods, in particular, in vitro cell selection, which is carried out on a callus culture. Therefore, at the first stage, it is necessary to develop in vitro technology for the rapid production of well-proliferating callus tissue. Purpose of the study. To study the effect of the mineral composition of the nutrient medium, photoperiod and endogenous polyphenols on the for-mation of the callus tissue of sequoia (Sequoia sempervirens (D.Don) Endl.) in vitro. Material and methods. The object of the study was Sequoia sempervirens (D.Don) Endl. plants. Callus tissue was obtained from segments of leaf blades and stem internodes, which were isolated from intact plants. Explants were cultured on MS and WPM nutrient medium containing BAP 2.5 mg/l and 2.5 mg/L 2,4-D. The localization of phenolic compounds was studied in the leaves, stems, apical buds of sequoia plants, as well as in callus tis-sue obtained on a nutrient medium with different mineral composition and under different lighting conditions. To do this, histochemical methods were used: for the sum of phenolic compounds, the material was stained with 0.08% Fast Blue reagent raster, a reaction with vanillin reagent in hydrochloric acid vapor was used to study the localization of flavans (catechins and proanthocyanidins). Results. It was found that the applied lighting mode had a significant effect on the intensity of callus tissue formation, its consistency and color. A well-proliferating callus tissue of light yellow color was obtained on two types of nutrient media with no light source. At a 16–hour photoperiod, a dark brown callus tissue was formed, which died during cultivation. As a rule, callus tissue was formed in those places where the localization of phenolic compounds was insignificant. It was found that in the initiated callus cultures grown on the studied nutrient media with no illumination, the content of cells with phenolic compounds was less than that of callus obtained on the same nutrient media, but with the presence of illumination. Conclusion. Sequoia plants have the ability to synthesize polyphenols, including the flavan series, which is preserved under in vitro conditions. The mineral composition of the studied nutrient media does not significantly affect the biosynthetic ability to form polyphenols in vitro cultures and their growth characteristics.

Features of the development of strawberry microplants in vitro

This article presents the results of a study of the effect of different sterilizers and nutrient media of different composition on the characteristics of the growth and development of garden strawberry (Fragaria x ananassa) microplants in vitro. To introduce a plant into a cell and tissue culture, two very important conditions must be met - to choose the right sterilizing solution, its concentration and processing time, and to choose the right balanced nutrient medium suitable for a particular culture. The studies were carried out on two varieties of garden strawberries: Favorit and White Swede. Sterilization of plant material was carried out with two sterilizers - 5% sodium hypochlorite solution and 1% silver nitrate solution. Studies have shown that the largest number of sterile explants was obtained by sterilizing the material with silver nitrate. Subsequently, viable meristems were planted on three nutrient media of different composition. According to the data obtained, it can be concluded that the MS nutrient medium with the addition of cytokinin 6-BAP at a concentration of 1 mg/L turned out to be the best in terms of the multiplication factor, the percentage of survival and the height of microplants..

Preliminary results of in-vitro laboratory propagation of barberry (Berberis L.)

The genus Berberis vulgaris (Berberis L.) is currently found in tropical, subtropical, temperate, and cold climate zones. There is little doubt that Southeast Asia is where the majority of Berberis vulgaris' interspecies biodiversity is found. Additionally, species are widely distributed, mostly in hilly areas, and, in terms of moisture requirements, they are mesophytic and xeromesophytic plants. In this article, experiments on in-vitro propagation of Berberis vulgaris (Berberis L.) seedlings were carried out in laboratory conditions. During the experiments, the time of sterilization of the explants obtained from the Berberis vulgaris plant was determined. Different concentrations of hormones and auxins were used in several nutrient mediums for growing in artificial nutrient medium. The results depicted that the highest rate was observed in the variant, where 2ml.BAP+0.5NAA+30g sucrose+7g accounted for 87% of plant rooting rate. It was observed that the rooting rate was slightly higher in the control variant of WPM-Woody plant medium-nutrient medium than the control variant of the MS nutrient medium, which was 17%.

Effects of different combinations of growth regulators on the in vitro growth parameters of Poncirus trifoliata L. (Raf.)

In this study, an in vitro propagation protocol of Poncirus trifoliata L. (Raf.) rootstock, which is one of the main rootstocks used in citrus, was created. Monocular node parts of the plant, which is the source of explant, were cultured in MS nutrient media containing BAP at dissmilar concentrations (0.5-1.0-2.0 mg/l). Afterward, it was observed that shoot appearance and tillering were achieved in the node explants cultured in MS nutrient medium comprising 1.0 mg/l BAP. The shoots were removed to a new culture in MS nutrient media containing NAA and IBA for rooting. While 30% rooting was achieved in the rooting medium, 62% of the rooted plantlets were acclimated to external conditions. 2 mg/l BAP and 2 mg/l silver nitrate (AgNO3) doses were supplied to the MS medium. The results of this study were obtained and a compound was found to the yellowing and leaf fall problem. As a result, in vitro propagation of Poncirus trifoliata, which is among the important citrus rootstocks, was tried, and its rooting ability was investigated in different nutrient media concentrations after shoots were obtained from different explants.

High Efficiency Plant Regeneration From Shoot Tip Explants Of Staurogyne Repens (Nees) Kuntze

A method for tissue culture of micropropagation of aquatic plant Staurogyne repens (Nees) Kuntze has been developed. Different phytohormones were investigated to optimize the growth of explants. The shoot tip explants of S. repens were placed in MS nutrient medium including BAP (0.30-1.50 mg/l) + TDZ (0.10 mg/l) + NAA (0.10 mg/l) and Kn (0.30-1.50 mg/l) + TDZ (0.10 mg/l) + NAA (0.10 mg/l). Shoot regeneration frequencies were changed as 77.77-100% and 66.66-100% in cultivation medium augmented with BAPTDZ- NAA and Kn-TDZ-NAA, respectively. In the growing media with BAP, the maximum shoot number (14.44 shoot/explant) was obtained in MS medium with 1.20 mg/l BAP + 0.10 mg/l TDZ + 0.10 mg/l NAA. In Kn applications, the highest shoot number (9.50 shoot/explant) was recorded in growing medium with 0.30 mg/l Kn + 0.10 mg/l TDZ + 0.10 mg/l NAA. The longest shoots (2.33 cm) was reached in nutrient medium with 0.90 mg/l Kn + 0.10 mg/l TDZ + 0.10 mg/l NAA. Regenerated plants rooting at 0.25 mg/l IAA have been effectively acclimated to aquarium conditions. Bangladesh J. Bot. 51(4): 697-704, 2022 (December)

Creation of seed breeding material of seed oats (Avena sativa L.) using embryo culture

The use of somaclonal seed oat samples in the breeding process allows to significantly expand the genetic variability of the experimental material. Callus was obtained by embryo culture from mature oat embryos sown on MS nutrient medium from 32.6% in No. 471–1117 with a concentration of 4 mg/l 2,4-D to 97.33% in No. 583–35 with a concentration of 2 mg/l. The regeneration ability of calli from different mature embryos of seeded oat offspring obtained on a nutrient medium with a concentration of 2,4-D in an induction medium of 2 mg/l ranged from 9.87 ± 2.42 in membranous offspring 479–1342 to 14.63 ± 5.52% in whole grains. As a result, regeneration lines of oat for obtaining seeds and further use in the breeding process were created.

Clonal Micropropagation of Cymbidium erythrostylum Rolfe

Cymbidium erythrostylum Rolfe is one of the most beautiful species of the genus cymbidium which is used as a cut flower or indoor plant. However, it was registered as a rare species even in its original habitat. This study was carried out to develop a protocol for in vitro propagation of C. erythrostilum. We used protosomes obtained by the asymbiotic germination of seed on 1/2 of Murashige and Skoog nutrient medium (MS) supplemented with 1 mg/L 2-Isopentenyl adenine (2iP) as objects of study. During the multiplication stage, the number of formed protosomes on a culture medium containing 0.8 mg/L 6-Benzylaminopurine (6-BAP) was higher (3.8 ± 0.2 protosomes). During the subsequent cultivation on ½ MS medium supplemented with 2 mg/L 6-BAP and 0.5 mg/L a-naphthaleneacetic acid (NAA), the highest numbers of shoots (4.00 ± 0.19 shoots/plantlet) and leaves (4.50 ± 0.14 leaves/plantlet) were obtained. At the rooting stage under in vitro conditions, the most effective was the use of ½ MS nutrient medium with the addition of 0.5 mg/L of indolyl-3-butyric acid IBA, 1 g/l charcoal, and 50 g/l banana puree. The obtained plants were successfully adapted to a substrate consisting of bark, perlite, and peat in a ratio of 1:1:1.