Stages Of Micropropagation Research Articles

Influence of culture medium composition on the micropropagation of Thymus tauricus Klokov et Des.-Shost.

Thymus tauricus Klokov et Des.-Shost. is a perennial dwarf shrub of the Lamiaceae family. This plant has anti-inflammatory, antispasmodic, analgesic and antiseptic properties. The use of biotechnological methods makes it possible to increase the efficiency of traditional methods of breeding and seed production. Our investigation aimed to study the influence of the culture medium composition on the explants morphogenesis at 1-2th stages of Thymus tauricus clonal micropropagation. When comparing three cytokinins, the best explants development was revealed on culture media containing kinetin. Both high vitrification rate of microshoots (31.2-90.2%) and formation of small shoots were observed on media supplemented with BAP or TDZ. The most effective culture medium at the introduction stage is MS with 1.0 mg/l kinetin and 1.0 mg/l GA3 or 1.0 mg/l kinetin. On this medium, on average, 8.4-10.1 microshoots per explant and shoots length of 1.9 cm were obtained. It was found that the optimal culture medium at the actually propagation stage is MS with 1.0 mg/l Kin or 1.0 mg/l kinetin and 0.5 mg/l IAA, on which the multiplication index 26.9-28.5 were obtained. Induction of rhizogenesis with a frequency of 30.2-94.4% and a root length of 1.6-3.0 cm was noted on culture media for the second stage of clonal micropropagation. The results of the studies are the basis for development T. tauricus clonal micropropagation method.

Ipomoea batatas (L.) Lam. cultivation in the conditions of light culture in vitro and ex vitro

Relevance. Currently, food products that include prebiotics, in particular, inulin, are particularly popular. Interest in this substance is justified by its valuable properties – it is a good immunomodulator, cleanses the body of toxins, radionuclides, "bad" cholesterol, promotes the assimilation of useful trace elements necessary for human life. Inulin is contained in plants such as jerusalem artichoke, chicory, as well as in sweet potatoes, the popularity of which is increasing every year. However, sweet potato plants are afraid of cold and frost-resistant. Therefore, the creation of new varieties and hybrids that are resistant to low temperatures is an urgent problem. Cellular biotechnology is aimed at solving this problem using methods of clonal microreproduction, cell selection, somatic hybridization, etc. For rapid reproduction and obtaining high-quality planting material, biotechnology methods are used, in particular, clonal micro-propagation. However, in this technology there are difficulties associated with poor adaptation of microclones to ex vitro conditions. This fact introduces an additional requirement for the selection of optimal rooting modes in vitro and ex vitro adaptation of microclones.Material and methodology. The aim of the work was to study the influence of cultivation conditions on in vitro rooting and ex vitro adaptation of I. batatas (L.) microclones. The object of the study was sweet potato microgears propagated in vitro. I. batatas micro-gears were cultured in vitro on a Murashige-Skug medium, differing by the type of auxins. The influence of red (R) and far red (FR) light on the shoots rooting in vitro and the adaptation of microclones ex vitro was studied.Results. It has been experimentally established that the cultivation of micro-gears on a medium containing indolyl butyric acid at a concentration of 0.5-1 mg/l and under conditions of illumination by LED lamps of red and far red light in equal amounts leads to the production of microclones with a well-developed root system and vegetative biomass. The use of an aeroponic installation at the last stage of clonal micro-propagation makes it possible to obtain high-quality planting material that can adapt well to open ground conditions.

Adaptation of microclones of blackberries to in vivo conditions

The results of studying the effect of mineral fertilizing on rhizogenesis and the development of aboveground organs of regenerant plants of blackberry thornless adaptable to in vivo conditions in the laboratory of selection, vegetable growing and horticulture, cloning “UNITS” Agrotechnopark “of Belgorod State Agrarian University are presented. Regenerated plants of thornless blackberry cultivar Agavam were adapted to in vivo conditions in a peat-perlite mixture with the addition of microelements and growth regulator root 16 days earlier than in the control. An active growth of the aboveground part and roots of regenerated plants of thornless blackberry was noted on the 21st day, in the control - on the 42nd day after the start of adaptation. By the end of the rooting stage on the 24th day, the regenerant plants formed an aerial part of two pairs of leaves 22 mm high and a developed root system - 37 mm. The mineral and hormonal composition of nutrient media for the cultivation of thornless blackberries has been optimized, an effective combination of physical and chemical factors at different stages of micropropagation has been determined, which enhance the proliferation of shoots and roots, and the dependence of the efficiency of adaptation of regenerated plants to in vivo conditions has been established. Along with traditional breeding methods, new opportunities for solving the problem of thorn-free blackberry varieties are provided, along with traditional breeding methods, which make it possible to accelerate the process of obtaining valuable planting material to provide the population and the processing industry with valuable berry products.

Regeneration from leaf explants of steppe cherry (Prunus fruticosa Pall)

The article represents data on morphogenesis from leaf explants of three steppe cherry genotypes, as well as the degree of somaclonal variability at in vitro and ex vitro stages, and in the field. It was revealed that a content of 6-benzylaminopurine, 4.43 μM, in combination with auxin, 0.5–0.6 μM, stimulates in the light the direct organogenesis in the tissues of the leaf base. This reaction was observed from 16.7 to 75.0% of explants, depending on the genotype. An equal 6-benzylaminopurine - auxin ratio (1: 1) led to the callus along with microshoots. Depending on the genotype, up to 30.0% of explants had such a mixed type of organogenesis. The mitotic index value in the apical leaflets differed depending on the day time. At the stage of micropropagation itself, an increase of the mitotic index was observed from 10 to 16 hours; at the stage ex vitro, no significant differences in the mitosis frequency were revealed within this time interval. No significant differences were found between the level of the mitotic index for plants obtained directly from leaves and those from buds. The mitosis passed without disturbances. No phenotypic changes in plant habit, shape and color of leaves were found.

Automation and digitalization of healthy fruit and berry plants reproduction

High-quality planting material is a condition for increasing productivity and labor productivity in the cultivation of agricultural products. High-quality planting material is material that is free from harmful pathogens. The process of plant healing requires significant time and material costs. However, with the proper organization, plant healing can be significantly accelerated and cheapened. The main condition for accelerating this process is maximum automation and digitalization of all stages of clonal micropropagation. The article shows the results of many years of work on the cultivation of healthy planting material. Variants of possible improvement of planting stock recovery are proposed with the introduction of the necessary hardware and software at specific stages of recovery and reproduction.

Optimisation of micropropagation protocols for temperate eucalypt hybrids in South Africa, with a focus on auxin transport proteins

Globally the demand for forestry resources is booming, and consequently innovative approaches to cultivate valuable hardwoods, such as eucalypts, are crucial. Although micropropagation offers a means to clonally propagate desirable cultivars, this technique is often variety-specific for eucalypts. Thus, recalcitrant genotypes are often excluded from micropropagation programmes as development requires large investments of time, money and expertise. In this study, a minimal maintenance medium was developed to reduce subculturing frequency (∼8 days) and to evaluate each micropropagation stage independently. Furthermore, a generic protocol for use across different varieties was developed by assessing the in vitro performance of three different Eucalyptus grandis x Eucalyptus nitens varieties (varieties 1–3) across various treatments. The purpose of this protocol was to ease the in vitro establishment of novel eucalypt varieties by providing a foundation for future protocol optimisation. During multiplication, medium containing 0.5 mg l−1 meta-topolin with 0.1 mg l−1 of indole acetic acid (IAA) induced the highest bud proliferation and shoot elongation for varieties 1 (59.1 ± 2.9; 1.2 cm ± 0.08), 2 (62.3 ± 2.7; 1.5 cm ± 0.09) and 3 (58.1 ± 3; 1.2 cm ± 0.07). Following multiplication, the rooting treatment that achieved the most consistent rooting percentages among varieties 1 (33.4%), 2 (43.5%) and 3 (34.3%) incorporated 0.029 mg l−1 of racemic-GR24 with 0.5 mg l−1 of IAA. Additionally, root vigour was assessed by measuring the root number and length, which varied considerably based on the variety and treatment in question. Overall, variety 2 was the most amenable to plant growth regulators and upon further investigation was found to possess equal expression levels of the auxin transporters PIN1 and AUX1. Conversely, the other varieties displayed unequal ratios of these transporters. Considering that auxins are principal media components, these expression profiles may serve as markers to identify eucalypt cultivars amenable to micropropagation.

The impact of copper oxide and silver nanoparticles on woody plants obtained by in vitro method

The article substantiates the necessity of studying behaviour of copper oxide and silver nanoparticles on forest cultures (downy birch 29-58 (Betula pubescens Ehrh.) and poplar ‘Pyramidal-osokoreviy Kamyshinsky’ (Poplus pyramidalis Roz. x Poplus nigra L.) in in vitro as well as in ‘soil-plant-microbiota’ to ensure stability of forest cultures in forest regeneration. The impact of nanoparticles on shoot regeneration and propagation processes was evaluated by introducing nanoparticles into Woody Plant medium. Differences in the influence of nanoparticles on the life processes of plants depending on their concentration and the stage of clonal micropropagation have been established. The results are demonstrated by a 15-25% reduction in the frequency of infection of poplar and birch explants as well as by an increase in their regenerating potential at the stage of introduction in tissue culture. When the nanoparticle solution is used in the soil substrate, a decrease in the number of diseased plants and an increase in their survival rate of 30% can be observed. The inhibitory effect of silver nanoparticles on some ecological and trophic groups of microorganisms has been established. These results can be used in the application of CuO and Ag nanoparticles in the biotechnology of clonal micropropagation of forest crops.

Влияние освещения на ризогенез ягодных растений при клональном микроразмножении

Introduction. Forest berry plants are popular on the food market and in pharmacy for their high nutritional and medicinal value. Plantations of forest berry plants can proliferate on unused lands, including depleted peatlands. Clonal micropropagation is the most effective method for obtaining large quantities of high quality planting material. Light-emitting diodes are highly effective for clonal micropropagation. The research objective was to study the effect of different spectral ranges on the process of root formation of forest berry plants in vitro.
 Study objects and methods. The research featured regenerant plants of half-highbush blueberry, arctic bramble, American cranberry, European cranberry, lingonberry, and Kamchatka bilberry of different cultivars. A set of experiments made it possible to study the effect of lighting type on the growth and development of the root system of forest berry plants in vitro using white fluorescent lamps, white spectrum LED lamps, and LED lamps with a combination of white, red, and blue spectra at the in vitro rooting stage of clonal micropropagation.
 Results and its discussion. The largest number (3.4–14.6 pcs.) and the maximum total length (10.0–156.9 cm) of roots were observed under LED lamps with a combination of white, red, and blue spectra. The effect was by 1.1–2.8 and 2.0–4.5 times higher than in the case of white-spectrum LED lamps, and by 2.3–7.0 and 3.3–14.9 times than in the case of fluorescent lamps. Variety and shape proved to have no significant effect on biometric indicators.
 Conclusion. LED lamps had a positive effect on the process of rhizogenesis of forest berry plants during clonal micropropagation. They appeared to be more effective than fluorescent lamps. The combination of white, blue, and red spectra increased the biometric parameters of plants at the stage of in vitro rooting.

EFFICIENCY OF MICROPROPAGATION OF SOME TAJIK SELECTION VARIETIES OF GRAPES IN IN VITRO CULTURE

An assessment of the micropropagation efficiency of Tajik selection varieties of grapes growing in the territory of the
 country was for the first time in Tajikistan given.
 During four transplantings different regenerative capacity in in vitro culture has been noted at the explants
 micropropagation stage of the explored varieties of grapes. The average coefficient of micropropagation of varieties is 2.5 and
 varies in the range from 1.2 to 3.2, which is apparently due to the varietal characteristics of grapes from the Tajik selection
 group. On the average for varieties of the Tajik selection group maximum propagation coefficient is noticed for the
 2nd transplanting.
 The average value of the propagation coefficient for all explants types of Tajik selection varieties of grapes differs a little
 between one another.

Unraveling Factors Affecting Micropropagation of Four Persian Walnut Varieties

Walnuts are considered recalcitrant to tissue culture, with a great genetic determinism on all stages of micropropagation; while other factors, also with great impact, become more complicated with the reproduction of newly realized varieties. In this research, a holistic approach aimed to determine the influence of genotype and the nutritive formulation throughout the whole cycle of micropropagation of four Persian walnut varieties (Juglans regia L.) was presented. During the in vitro establishment it was determined that besides genotype and culture medium, the effect of collection season and the likely interaction amongst factors had a great influence on the successful introduction of all four genotypes. However, all cultures were affected by a deep decay, being necessary the introduction of ethylenediamine di-2-hydroxyphenyl acetate ferric, as iron source, and Phloroglucinol in both Murashige and Skoog (1962) and the corrected Driver and Kuniyuki (1987) formulations. These modifications allowed the stabilization of cultures, maintaining thereafter a steady quality. Either proliferation, rooting and ex vitro survival of four clones were affected by the culture medium, obtaining the best results with the corrected Driver and Kuniyuki (1987) formulation. Finally, in vitro plants produced from all clones were acclimated with high survival rates (75.9–91.1% for the best culture medium), depending of clone and the culture medium used. Microsatellite analysis showed that micropropagated plants maintained the same genetic profiles of their corresponding donor trees. These results might contribute to deepening of the understanding of factors that determine the success of micropropagation of walnuts, and the extents of its influence; whereas, it sets the basis for the commercial micropropagation of all four clones.

Influence of Light Conditions and Medium Composition on Morphophysiological Characteristics of Stevia rebaudiana Bertoni In Vitro and In Vivo

We investigated the influence of different conditions (light composition and plant growth regulators (PGRs) in culture media) on the morphophysiological parameters of Stevia rebaudiana Bertoni in vitro and in vivo. Both PGRs and the light spectra applied were found to significantly affect plant morphogenesis. During the micropropagation stage of S. rebaudiana, optimal growth, with a multiplication coefficient of 15, was obtained in an MS culture medium containing 2,4-epibrassinolide (Epin) and indole-3-acetic acid (IAA) at concentrations of 0.1 and 0.5 mg L−1, respectively. During the rooting stage, we found that the addition of 0.5 mg L−1 hydroxycinnamic acid (Zircon) to the MS medium led to an optimal root formation frequency of 85% and resulted in the formation of strong plants with well-developed leaf blades. Cultivation on media containing 0.1 mg L−1 Epin and 0.5 mg L−1 IAA and receiving coherent light irradiation on a weekly basis resulted in a 100% increase in the multiplication coefficient, better adventitious shoot growth, and a 33% increase in the number of leaves. S. rebaudiana microshoots, cultured on MS media containing 1.0 mg L−1 6-benzylaminopurine (BAP) and 0.5 mg L−1 IAA with red monochrome light treatments, increased the multiplication coefficient by 30% compared with controls (white light, media without PGRs).

СОВЕРШЕНСТВОВАНИЕ КЛОНАЛЬНОГО МИКРОРАЗМНОЖЕНИЯ ЯГОДНЫХ КУЛЬТУР

The aim of the study is to optimize the conditions for in vitro cultivation of blue honeysuckle, raspberry and strawberry. The work was carried out in 2012-2020. The Murasige-Skuga medium (1/2 MS) was the control for all cultures for the initiation of explants. Additionally, we used a modified nutrient medium 1/2 MS with a reduced NH4 content by 15 % compared to the base MS; and Woodi Plant Medium (1/2 WPM) for honeysuckle; for raspberries - Quoirin-Lepoivre (1/2 QL) and 1/2 Anderson; for strawberries - 1/2 MS. For micropropagation and rooting, the following media were used: honeysuckle - modified MS and WPM; raspberries - QL and Anderson; strawberries - MS modified by Siliplant and Boksyu; control for all - MS. The following growth regulators were added to the optimal each culture a nutrient medium: 6-benzylaminopurine (6-BAP), gibberellic acid (GA), waste products of the large wax moth larvae, indolyl-3-butyric acid (IBA), Siliplant, EcoFus, HB-101. The effect of LED-phytoirradiators with a combination of red, blue and white light in the spectrum 2: 1: 1, 1: 1: 1, 2: 1, respectively, and LED-irradiators with a changing spectrum and flashing were studied at the stages of micropropagation and rooting in all cultures. The survival rate of honeysuckle explants on 1/2 WPM medium was 62.2 % (control 27.9 %). The highest reproduction factor of 5.1 (control 2.6) was achieved when using LED 2 red : 1 blue : 1 white on MS modified + 6-BAP 1.0 mg/L + kinetin 0.5 mg/L, and high rooting rate of honeysuckle 89.0 % (76.0 % k) was achieved on MS modified + IBA 0.5 mg/L. Cultivation of red raspberries on QL + 6-BAP 1.0 mg/L + GA 0.5 mg/L and LED irradiation 2 red : 1 blue : 1 white provided a reproduction factor of 5.3 (control 2.7), addition of IBA 0.5 mg/L + HB-101 100 μL/L in QL and LED irradiation 1 red : 1 blue : 1 white contributed to 100 % rooting. The addition of 6-BAP 1.0 mg/L + IBA 0.2 mg/L + GA 0.5 mg/L in QL and LED lighting 1 red : 1 blue : 1 white increased the reproduction factor of remontant raspberries by 1.6 times (from 2, 6 to 4.1), and the use of QL + IBA 0.5 mg/L + HB-101 50 μL/L and LED 2 red : 1 blue : 1 white increased its rooting ability to 96 % (control 67 %). LED irradiation with a changing spectrum during cultivation of garden strawberries on MS + Siliplant + EcoFus at 0.5 ml/L provided a reproduction factor of 5.9 (control 3.8), and the reproduction factor of remontant strawberries on MS + HB-101 100 μl/L was 7.4 (control 5.6). The addition of IBA 0.5 mg/L + HB-101 100 μL/L to the MS promoted the rooting of garden strawberries of 100 % when using a LED irradiator with a changing spectrum, and remontant strawberries – with a blinking LED irradiator

ПОЛУЧЕНИЕ ВЫСОКОКАЧЕСТВЕННОГО ПОСАДОЧНОГО МАТЕРИАЛА ЛЕСНЫХ ЯГОДНЫХ РАСТЕНИЙ МЕТОДОМ КЛОНАЛЬНОГО МИКРОРАЗМНОЖЕНИЯ ДЛЯ РЕКУЛЬТИВАЦИИ ВЫРАБОТАННЫХ ТОРФЯНИКОВ

The results of studies on the improvement of technology for producing highquality planting material of half-high blueberry and Arctic bramble by the method of clonal micropropagation are presented in the current paper. Creation of forest berry plantations in peat extraction areas allows reducing environmental damage and significantly increasing the efficiency of the timber industry. In recent decades, there has been an increasing interest in the creation of forest berry plantations on drained and cutover peatlands in Russia and other countries. It is necessary to use varietal planting material for the successful cultivation of forest berry plants on an industrial scale. Clonal micropropagation is the most effective of the vegetative methods for obtaining planting material, which allows receiving a huge amount of healthy planting material all year round in the conditions of a small laboratory area. Chloride-free ecosterilizer and bleaching agent based on sodium hypochlorite “Belizna” with an exposure of 15 and 20 min showed high efficiency in sterilization of explants of half-high blueberry and Arctic bramble. The highest viability of explants of the studied forest berry crops was observed when sterilized with a 0.1 % mercuric chloride solution and 15 min exposure, and its sharp decrease at 20 min exposure. At the stage of micropropagation, with an increase in the concentration of cytokinin 6-BAP from 0.5 to 1.0 mg/L on the nutrient Woody Plant Medium the number of shoots in regenerated plants of half-high blueberry (Northcountry and Northblue cultivars) and Arctic bramble (Anna and Sofia cultivars) increased. The effect of the concentration of IBA-derived auxin on the number and length of roots of regenerated plants was observed at the in vitro rooting stage.

Bazı Turunçgil Melezlerinin in vitro Koşullarda Mikroçoğaltım ve Köklenme Performanslarının Araştırılması

Turunçgiller yetiştiriciliği yapılan en önemli ve yüksek tür çeşitliliğine sahip meyve gruplarından biridir. Taze olarak tüketilmesi ve farklı endüstrilerde ham madde olarak kullanılması turunçgillerin önemini artırmaktadır. Bu nedenle turunçgillerde yeni çeşit ve anaç geliştirme çalışmaları oldukça önemlidir. Melezleme çalışmaları sonucunda elde edilen değerli bitkilerin çoğaltılması için bitki doku kültürü çalışmaları güçlü bir alternatiftir. Bu çalışmada, Sunki mandarin X Rubidoux üç yapraklı melezlerinin in vitro koşullarda mikroçoğaltımında genotip ve farklı besin ortamlarının (Murashige and Skoog Medium -MS, Woody Plant Medium -WPM, Rugini Olive Medium -ROM) etkisi incelenmiştir. Ayrıca bitkilerin köklendirilmesi üzerine genotip ve farklı bitki büyüme düzenleyicilerin (indol bütirik asit - IBA ve naftalen asetik asit - NAA) etkisi araştırılmıştır. Elde edilen köklü bitkiler dış koşullara alıştırılmıştır. Çalışmada kullanılan genotiplerden mikroçoğaltım aşamasında en başarılı genotipler SR-60 ve SR-75 olurken, köklenme aşamasında en başarılı genotipler SR-41, SR-47 ve SR-60 olmuştur. Besin ortamlarının mikroçoğaltım üzerine etkisi istatistiksel olarak önemsiz bulunmuştur (P<0.05). Köklenme denemelerinde bitki boyu ve kök uzunluğu için en başarılı bitki büyüme düzenleyici IBA, kök sayısı için en başarılı bitki büyüme düzenleyici ise NAA olarak belirlenmiştir.

Paphiopedilum insigne Morphological and Physiological Features during In Vitro Rooting and Ex Vitro Acclimatization Depending on the Types of Auxin and Substrate.

To obtain healthy and good quality plants from in vitro cultivation, it is necessary to produce plantlets with well-developed rooting systems because they must undergo acclimatization, a final and a very difficult stage of micropropagation. In the present research, the effect of auxins NAA, IAA and IBA in concentrations of 0.5; 1; 2.5 and 5 mg·dm−3 on the Paphiopedilum insigne in vitro rooting was studied, and it was noted that 1 mg·dm−3 of IAA or IBA enabled the obtaining of a lot of rooted and good quality plantlets. The subsequent influence of the two most advantageous auxins on the acclimatization of plantlets in different substrates (sphagnum moss, sphagnum moss + substrate for orchids, substrate for orchids, substrate for orchids + acid peat) was tested, in the means of morphological features of plants and their physiological parameters, i.e., chlorophyll fluorescence (FV, Fm, Fv/Fm), stress enzyme activity (catalase, ascorbate peroxidase), and water balance. Considering all the tested features, it might be stated that the best results were obtained when explants were rooted in vitro in the presence of 1 mg·dm−3 of IAA and then planted ex vitro in substrate for orchids.

Clonal micropropagation of bog cranberry (Vaccinium oxycoccos L.) in the Udmurt Republic

The paper presents the results of 2018-2019 research on improving the technology of growing planting material of bog cranberry (Vaccinium oxycoccos L.) of Krasa Severa, Severyanka, Virussaare varieties on the basis of in vitro. Studied was the effect of the concentrations of growth regulators in the composition of the nutrient medium according to Anderson's recipe on the reproduction and subsequent rooting of micro cuttings, as well as the duration of cultivation and adaptation of micro plants depending on partial pruning of shoots. It has been established that at the stage of introduction into in vitro culture, sterilization of explants with 33% hydrogen peroxide in an exposure of 5-8 minutes with washing in 5 portions of sterile distillate gives 60-80 % of viable shoots. The optimum phase of plant development for the successful introduction of in vitro culture is the swelling of buds. Cultivation of micro cuttings was carried out in a light room at a temperature of 25±2 °С, a photoperiod of 16 hours. The duration of each subculturing was 30-60 days. For the stage of actual micropropagation on Anderson's nutrient medium, an increase in the dose of cytokinin 6-benzylaminopurine (6-BAP) from 0.2 to 0.5 mg/l and an increase in the duration of cultivation from 30 to 60 days contributed to a significant increase in the multiplication factor on average for the tested cranberry varieties.According to the efficiency of micropropagation, the varieties Virussaare and Krasa Severa were distinguished – 9.3-12.0 pcs/stalk, respectively. At the rooting stage, the use of a root-forming reagent of indolyl-3-acetic acid (IUK) in doses of 0.2, 0.5 and 1.0 mg/l in the composition of Anderson's nutrient medium did not affect the quality of root formation and the length of shoots of Virussaare micro-plants. No significant varietal differences in the root-forming ability of microcuttings were found. The tendency of better rooting of micro cuttings was observed in the Virussaare variety (90.3 %) compared to the Severyanka (85.7 %) and Krasa Severa (79.3 %) varieties. Micro plants of the Krasa Severa cultivar were characterized by the longest shoots, the total number of roots was less, but their length was longer in comparison with other cultivars. For the adaptation stage, a substrate from a mixture of lowland peat and sphagnum moss was used (1:1). The efficiency of adaptation of micro plants of cranberry varieties when cutting the tip of the shoots was 100 %. Pruning of micro plants shoots contributed to the formation of more side shoots and better development of the aboveground part of the plants.

The effect of humic acid (HA) and zinc oxide nanoparticles (ZnO-NPS) on in vitro regeneration of date palm (Phoenix dactylifera L.) cv. Quntar

Date palm (Phoenix dactylifera L.) micropropagation still faces many problems, such as reduced growth and development of callus, low multiplication efficiency and low rooting rate. The effect of Humic Acid (HA; 0, 0.5, 2.5, and 5.0 mg L−1) and Zinc oxide nanoparticles (ZnONPs; 0, 50, 75 and 150 mg L−1) at different stages of micropropagation (i.e., callus establishment, shoot multiplication and rooting) was studied in date palm cv. Quntar. Media supplemented with 2.5 mg L−1 HA in combination with 50 m L−1 ZnO-NPs gave the highest callus induction (274 mg/jar), the highest percentage of callus producing buds (83.34%), and average bud formation (14.2) per callus. The macronutrients Nitrogen (N), Phosphorus (P), Potassium (K), Sulfur (S) as well the micronutrient Zinc (Zn), significantly increased in in vitro shoots regenerated in medium containing of 2.5 mg L−1 HA and 50 mg L−1 ZnO-NPs compared to other treatments. The addition of HA and ZnO-NPs to the medium was most effective in root regeneration and the number of roots per shoot, where the best result (83.34%, 4.4 roots per shoot, respectively) was obtained using 2.5 mg L−1 humic acid and 75 mg of L−1 zinc oxide (ZnO-NPs), compared with other treatments. These results also indicate that using 2.5 mg L−1 HA combined with 75 m L−1 ZnO-NPs as a supplement can increase antioxidant enzymes CAT and POD compared with other treatments. The data revealed that maximum chlorophyll content of shoots occurred in a medium supplemented with 2.5 mg L−1 HA and 75 mg L−1 ZnO-NPs. Furthermore, we found that the combined application between 2.5 mg L−1 HA + 75 mg L−1 ZnO-NPS resulted in the highest shoots content of endogenous IAA (3.644 µg.kg−1), compared with other treatments. We have established a highly efficient micropropagation system for date palm by using (HA) and zinc oxide nanoparticles (ZnO-NPS). The initiation, broad optimization, and comprehensive phytochemical studies of tissue cultures were elaborated.

Aftereffect of long-term deposition of raspberry micro-plants in a light room on nutrient media with the addition of Superstim drug modifications on their rhizogenesis and adaptation

When the clonal micropropagation of raspberry remontant the sorts of Briliantovay and Hercules) at the stages of in vitro rhizogenesis and adaptation to non-sterile conditions, the aftereffect of depositing micro-plants in a light room was evaluated when adding Superstim-1 and Superstim-2 preparations in small and ultra-small doses to the nutrient medium. Analysis of effective concentrations during deposition and aftereffect studies at all further stages of clonal micropropagation showed the advantage of the preparation Superstim-1 in concentration for the diamond variety 1 × 10-9 %, for the Heracles variety 1 × 10-6 %. When you Deposit in the culture room, micro plants remain viable without signs of necrosis and chlorosis for 10-12 months, during the remediation of the propagation coefficient is 3-5 times higher than those of control, on the rhizogenesis 20-30 days reduce the duration of the period of subcultivation, and the best options 100% accustomed at the stage of adaptation to non-sterile conditions in 2 - 2.5 times superior to the control indicators.

Biotechnology for the production of Iris spuria L. plant material with a specified content of extractives

The study was aimed at the establishment of stock plant material of Iris spuria L. (Art And Soul) through biotechnological methods and its primary phytochemical analysis. For the first time, a series of studies aimed at multiplying plant materials Iris spuria L. (Art And Soul) resulted in the development of culture media and cultivation schemes. In the stage of micropropagation as such, the most optimal content in culture media was found to be 2.5-5.0 µm BAP. To ensure a more complete realization of morphogenetic potentials, it is necessary to alternate phytohormone and hormone-free media. In this case, L-glutamine and adenine sulfate should be added to hormone-free media in the amount of 100 mg/l (MS+100 mg/l L-glutamine+100 mg/l adenine sulfate). Qualitative and quantitative compositions of extractives removed in a Soxhlet apparatus from the technological plant material of Iris spuria depends on the solvent polarity. The amount of extractives increases with accelerated solvent polarity (96% ethanol → 60% ethanol → water). In this case, phenols, condensed and hydrolyzable tannins, alkaloids, glycosides were extracted. The study proves the relationship between the accumulation of quercetin and the hormonal composition of culture media, which allows for the regulation of these polyphenols being accumulated in the production of the plant material of Iris spuria L. (Art And Soul).

Some aspects of clonal micropropagation of Amelanchier Medik. genus representatives

The article presents studies of organogenesis peculiarities of varieties belonging to various species of Amelanchier Medik. genus in in vitro culture at the stages of micropropagation, rooting and depositing. Mineral composition and growth regulator for active adventive microshoots formation were determined. The optimal auxin source for rhizogenesis of various Amelanchier alnifolia varieties has been established. Conditions for explants' preservation in aseptic conditions for 12 months have been revealed. Different effects of cytokinin type substances on shadberry regeneration potential under aseptic conditions were noted. Analysis of regenerants' morphometric indicators showed the effectiveness of using 6-benzylaminopurin (BAP) in the concentration of 1.0 mg/L for shadberry varieties' clonal micropropagation and indolebutyric acid (IBA) at 1.0 mg/L concentration for microshoots' rooting.