Indolylbutyric Acid Research Articles

In vitro propagation of the rare fern Polystichum craspedosorum (Maxim.) Diels. via green globular bodies

Polystichum craspedosorum (Maxim.) Diels. is a rare species of fern listed in the Red Data Book of the Amur Region. Cultivation of ferns in vitro via green globular bodies (GGBs) is a promising direction for the conservation and reproduction of ferns. The objective of the study was to estimate the efficiency of different media on GGB propagation and sporophyte regeneration of P. craspedosorum. Two media options were effective for GGB propagation and sporophyte regeneration: 1/2MS without ammonium nitrate, vitamins and plant growth regulators (variant 1) and 1/2MS without plant growth regulators (variant 2), both of which were also supplemented with 2 % sucrose and 0.8 % agar, and adjusted to pH 5.8. At the same time, media containing a single cytokinin (0.5 mg/l 6-benzylaminopurine) and a single auxin (0.5 mg/l indolylbutyric acid) were less effective for GGB propagation and sporophyte regeneration. The total number of initiated GGBs was 313.4 and 275.8 as a mean per explant after two months in vitro culture for variant 1 and 2, respectively. The number of GGBs with developed sporophytes was 187.1 and 158.4, the sporophyte regeneration rate was 59.7 % and 57.4 %. The sporophytes survival rate was 85 %.under ex vitro conditions.

КАЛЛУСООБРАЗОВАНИЕ КАК ФИЗИОЛОГИЧЕСКИЙ ПРИЗНАК НЕДОСТАТКА АУКСИНА В СОСТАВЕ СТИМУЛЯТОРА РИЗОГЕНЕЗА (НА ПРИМЕРЕ УКОРЕНЕНИЯ IN VIVО ЧЕРЕНКОВ ХВОЙНЫХ КУЛЬТУР)

Characteristics of the effectiveness of rooting of coniferous crops are the percentage of rooting, the number of roots formed and their length. Callus formation often accompanies the rooting process, but research data on the relationship of this process with the effectiveness of rooting is contradictory. In the conditions of film cuttings equipped with heated substrate, studies were carried out on the relationship between the rooting of cuttings of coniferous crops and callus formation. To stimulate the rooting of semi-lignified cuttings of Juniperus scopulorum S. cultivar Skyrocket, growth powders Chrysatop and Rizapon 1% containing indolyl-3-butyric acid (IBA) in different concentrations were used. In the control treatment without a stimulant, all cuttings formed callus without rooting; when the bases of the cuttings were dusted with Chrysatop containing 0.25% IBA, callus formation decreased by 3.8 times, and the survival rate was 76%. In the variant with Rizapon powder containing 1% IMC, no callus formation was observed at all, the survival rate reached 81.4%. To stimulate the rooting of green cuttings of Juniperus scopulorum S. variety Blue Arrow, aqueous solutions of potassium salt IMC (IMC-K) were used in the concentration range from 0 to 150 mg/l when the bases of the cuttings were soaked for 16 hours. In the concentration range from 0 to 100 mg/l, an increase in survival rate by 1.89 and 3.87 times was observed, respectively, callus formation decreased by 3.38 times. To stimulate the rooting of semi-lignified cuttings of Thuja occidentalis L. variety Smaragd, Juníperus sabina L. species form and green cuttings of Thuja occidentalis L. variety Brabant, Kornevin growth powder containing 0.5% IBA was used. To obtain a concentration gradient, the powder was diluted with talc. Against the background of high survival rate of cuttings of Thuja occidentalis L. of both varieties – 82.85–96.9%, callus formation decreased from 92.9% in the control variant to 6.3% in the variant with pure Kornevin in cuttings of the Smaragd variety and from 71.88% in the control variant to 19.1% in variant Kornevin:talc 1:1 in cuttings of the Brabant variety. When rooting cuttings of Juniperus sabina L. with an increase in the concentration of IBA in the powder from 0 to 0.25% IBA, the survival rate increased from 10.71 to 89.29%, and the formation of callus on the cuttings decreased from 44.59 to 0%. The data obtained indicate that calus formation can serve as a sign of a lack of exogenous auxin in the rooting stimulator and, accordingly, an increase in the auxin content, all other things being equal, within the effective range is one of the ways to increase the percentage of survival of cuttings of coniferous crops.

The influence of mineral fertilizers and plant growth regulators on the development of clone rootstocks of apple trees 54-118 in container growing technology

When replicating apple tree planting material in protected soil, technologies for growing plants in container culture using an organomineral substrate, long-acting mineral fertilizers and plant growth regulators are widely used. The purpose of the research was to establish the effectiveness of the combined use of plant growth regulators and long-acting fertilizers when growing clonal apple rootstocks 54-118 in container technology in protected ground. The experiments were carried out in 2021-2023 according to generally accepted methods. The article presents the results of a study of the effect of joint use of OsmocotePRO long-acting fertilizer for 3-4 months. (17-11-10+2MgO+TE) and plant growth regulators indolyl-3-butyric acid (IBA) and gibberellic acid (GA3) on plant growth and development. These technology elements can be used both when obtaining planting material for planting mother plants of clonal rootstocks, and for obtaining seedlings.

Rhizogenesis of rocket (Eruca sativa Mill.) and wall-rocket Diplotaxis tenuifolia (L.) DC.

Purpose. To test the efficiency of the plant growth regulating substances at different concentrations for stimulating the rhizogenesis of rocket (E. sativa) and wall-rocket (D. tenuifolia). Methods. The experiment was carried out at the Biotechnology Laboratory of the Institute of Bioenergy Crops and Sugar Beet of the National Academy of Agrarian Sciences in 2020–2022. Rocket varieties ‘Znahar’, ‘Lybid’, ‘Zlat’, ‘Silvetta’ and wall-rocket variety ‘Liudmyla’ were used in the study. Seeds were examined for their quality indicators, sterilized with a solution of 35% laundry detergent and ethanol and seeded in vitro. Sterile seedlings were transferred to the Murashige-Skoog (MS) medium supplemented with benzylaminopurine (BAP) for propagation. Naphthylacetic acid (NAA), indolyl-3-acetic acid (IAA), indolyl-3-butyric acid (IBA) and gibberellic acid (GA) were added to the nutrient medium according to the MS prescription in different concentrations. Results. The introduction of IBA (0.8 mg/l) and GA into the nutrient medium, regardless of the concentration, resulted in the longest root system in all treatments. The studied modification made it possible to obtain root length on the 9th day of cultivation from 7 to 64 cm and on the 14th day from 10 to 70 cm. On the 14th day of cultivation, in the plants grown under the concentration of GA >1.0 mg/l, the root not only the elongated and browned, but reached 70 cm in length. For the introduction of IBA into the medium previously supplemented with GA, on the 9th day of cultivation, negative effect was noted in all treatments, namely the elongation of internodes and central root. On the 14th day, vitrification of shoots and significant elongation of both central and lateral roots occurred in all treatments. The central root reached 70 cm in length, which is not advisable during the adaptation period. The results of the experiment indicate that on the 21st day, the longest root system (12 cm and 42 cm) was obtained in the rocket variety ‘Zlat’ (0.1 mg/l GA and 1.5 mg/l GA, respectively) and the shortest root length (6 cm and 21 cm) in the wall-rocket variety ‘Liudmyla’ (0.1 mg/l GA and 1.5 mg/l GA, respectively). Importantly, the root length of ‘Zlat’ variety on the 21st day of cultivation, compared to the 14th day, at a concentration of 0.1 mg/l increased by only 3 cm, while in the 'Liudmyla' variety by 2 cm. Considering this, it is impractical to carry out cultivation up to 21 days. The lowest root length was obtained with the use of IBA and GA: rhizogenesis occurred more slowly than in previous treatments, and on the 14th day the root length was in the range from 3 to 20 cm. Conclusions. The longest root system in all studied treatments was obtained in variety ‘Zlat’, while shortest in ‘Liudmyla’. The optimal concentrations of NAA + IBA + GA for the cultivation of the studied rocket and wall-rocket varieties are 0.3 and 0.5 mg/l.

Compositions of Fungicides with Growth Regulators which Reduce the Retarding Effect of Crop Protectants

One of the reasons for the decrease in the yield of crops is the retardant effect of fungicides. In addition, many fungicides have a toxigenic effect on the mycelium of the fungus. The combination of a fungicide and a plant growth stimulant in a complex preparation can help to reduce the level of the negative effect of the pesticide. It is proposed to introduce individual growth regulators and their combination into the composition of the seed protectant in doses of 50, 100, 200 g/l. The length and the weight of the root and shoot of seedlings under the influence of the seed protectant were less than in the control, and the weight of the endosperm of grain was greater. Under the influence of the seed protectant and gibberellic acid, the length of the shoots corresponded to the control, but their weight was decreased. Indolylbutyric acid, added to the seed protectant, contributed to the effective consumption of spare substances of the endosperm of grain and root growth. Under the influence of the protectant and diphenylurea, the length and the weight of the shoot increased significantly in a balanced manner. The composition of the protectant and the named growth regulators in minimal doses affected the growth of the roots and shoots in the best way of all variants. Preference should be given to low doses of phytohormones and their combination.

ОЦЕНКА БИОМЕТРИЧЕСКИХ ПОКАЗАТЕЛЕЙ КЛОНОВЫХ ПОДВОЕВ ГРУШИ И АЙВЫ, ПОЛУЧЕННЫХ С ПОМОЩЬЮ ЗЕЛЕНЫХ ЧЕРЕНКОВ

Pear and quince belong to different botanical species, so most pear varieties have partial or complete incompatibility with quince. Quince is also a relative of pears, they have the same diseases and pests, so preventive treatments need to be carried out several times a season. Quince is not only planted next to a pear, it is also used as a rootstock for growing dwarf pear varieties. But the pear is more difficult to root without special plant growth regulators than the quince. Pear culture is very difficult to reproduce without specialized plant growth stimulants. Currently, there are a lot of different preparations for rooting various fruit and berry crops. Growth stimulants strengthen the root system of plants, accelerate the flowering process, promote earlier formation of fruits, increasing their size. Clonal rootstocks are plants obtained by vegetative propagation from green or lignified cuttings. The article presents the results of research on the use of various plant growth stimulants, with the help of which clonal rootstocks of pears and quinces were rooted and further studied. As substances stimulating the processes of root formation, an aqueous solution was used: indolyl butyric acid (BCI) - 50 mg/l for 24 hours, epin-extra - 1.0 mg/l for 24 hours, succinic acid - 200 mg/l for 24 hours. Water was used as a control. It was found that the greatest rooting result of green cuttings treated with β-indolyl-3-butyric acid was characterized by Northern quince (66.7%), also pear forms PG 12 (k) (57.6%), PG 2 (58.4%), PG 17-16 (59.3%). When processing varieties, forms of pear and quince with succinic acid, Northern quince (60.0%) and pear forms PG 17-16 (58.7%), PG 12 (k) (55.7%), PG 2 (56.7%) had the greatest rooting result. When using epin-extra, the greatest rooting results were: Northern quince (55.0%), pear forms PG 12 (k), PG 2, PG 17-16, (50.0%, 52.3% and 54.0%, respectively). Without treatment with plant growth stimulants, the best result was observed in Northern quince (51.7%). When treated with plant growth stimulants with β-indolyl-3-butyric acid, succinic acid, epin-extra and without the use of plant growth stimulants, Northern quince had the highest plant height, the diameter of the conditional root neck, the number of roots, and the length of the roots.

Use of Plant Regulators for Activation of Antioxidant Enzymes in Basil Plants under Water Deficit Conditions

Basil is susceptible to biotic or abiotic stress, negatively interfering with growth and production. Thus, the objective of this work was to evaluate the physiological effects of the application of plant regulators in basil plants that suffer from water deficit. The experiment was conducted in a randomized block design (RBD) in a 2 × 4 factorial scheme, including plants that were subjected to water stress and those that were not. In addition, plants also received five doses of Stimulate® composed of indolylbutyric acid (IBA) + gibberellic acid (GA3) + kinetin (Kt) with four repetitions each. The experiment was evaluated through the biochemical analyses of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and lipid peroxidation performed 20, 35, and 50 days after transplanting (DAT). The mixture of plant regulators attenuateds the effects through the increasing activities of these enzymes. The plants that received the highest dosages (9 and 12 mL L−1) offered the best protetion. Parameters of growth measures such as number of leaves and leaf area also showed significant responses regarding the application of the plant growth regulators. The use of a mixture of plant regulators, despite satisfactory results, does not make basil economically viable because it presents inaccurate results regarding its use.

Effect of different concentrations and combinations of some plant growth regulators on Punica granatum anther culture

Plant growth regulators (PGR), also known as plant hormones, are a variety of chemical compounds that have a significant impact on the development and differentiation of plant cells. This work aimed to study the effect of various combinations of Plant Growth Regulators (PGRs) for example [6-Benzyladenine (BA), 2,4-dichlorophenoxyaceticg acid (2,4-D), Naphthalene acetic acid (NAA) and indol-3-ylbutyric acid (IBA)] on callus induction and shoot regeneration from anther culture of S1 progenies resulted from Manfalouty cultivar open pollinated. The results showed that highest value of percentage of callus induction was 10.6% on MS medium supplemented with 1 mg / l of NAA and 2 mg / l of BA also highest value of percentage of shoot formation 60 % on MS medium supplemented with 3 mg/ l of NAA and 2 mg /l of BA.

Micropropagação de Trichopilia suavis Lindl. & Paxton

Abstract Trichopilia suavis is one of the important representatives of the genus Trichopilia, which can be used as a potted plant for the interior design. also, its beautiful inflorescences are added unusual element to bouquet compositions and actively used in the production of perfumes. This study was carried out in order to develop and improve micropropagation method for mass clonal production of T. suavis through protocorm culture. Half-strength Murashige and Skoog culture medium (½) has been modified by adding different organic components and growth regulators. The protocorms obtained from seeds germinated by asymbiotic method were used as explants. The results showed that the optimal treatment for formation of new protocorms (7.50 ± 0.7 units per protocorm) was obtained by culture on ½ MS medium with 0.5 mg L-1 of 6-Benzylaminopurine (6-BAP) and 100 mL L-1 of coconut water. At the stage of rooting, the highest number and length of roots (7.0 ± 0.45 units per plantlet, 2.64 ± 0.18 cm) were obtained through subsequent cultivation of plantlets on ½ MS medium with 0.5 mg L-1 indolyl-3-butyric acid (IBA), 1.0 g L-1 charcoal with the addition of 50 g L-1 of banana puree. The obtained seedlings were successfully adapted in a substrate consisting of bark, perlite and peat in a ratio of 1:1:1.

THE EFFECT OF INDOLYLBUTYRIC ACID ON THE REGENERATION OF MAIZE HYBRIDS IN VITRO

Еру Optimization of the regeneration potential in vitro for maize (Zea mays L.) hybrids with the participation of perspective inbreds is an actual task for biotechnological maintenance of this important crop breeding process.
 The purpose of the research was the investigation of the effect of the auxin phytohormone — indolylbutyric acid (IBA) on the in vitro regeneration capacity of maize hybrids. The task of the work was to determine the effect of IBA in concentrations of 0.1 and 0.2 mg / L on the total regeneration frequency from type II callus tissue, as well as on the number of regenerated plants per 100 calli. The material of the study was 90-day type II callus tissue of maize hybrids between inbreds DK633, DK3070, DK236 and DK267 of Lancaster germplasm from the Dnipro breeding program and inbreds with high regeneration capacity A188, Chi31 and PLS61. The method of cell, tissue and organ culture in vitro was used. The initiation of plant regeneration from callus tissue was performed on the mineral MS medium, supplemented with 100 mg / L inositol, 100 mg / L casein hydrolysate, 690 mg / L L-proline, 20 g / L sucrose, 6 g / L agar, and 0.1 or 0.2 mg / L IBA.
 It was found that IBA significantly affected the maize hybrids’ regeneration capacity. With 0.1 mg / L IBA in the medium, the total regeneration frequency increased up to 32.1 ± 7.7 %, compared to the control without IBA (15.6 ± 5.3 %) and to 0.2 mg / L IBA (17.0 ± 5.5 %). IBA (0.1 mg / L) contributed also to the increase of the number of regenerated plants up to 19.5 ± 4.3 pcs/100 calli, compared with their number without IBA (9.7 ± 4.3 pcs/100 calli) and with 0.2 mg / L IBA (6.5 ± 3.6 pcs/100 calli). The genotype of callus tissue significantly affected the regeneration ability of the hybrids. The highest number of regenerated plants per 100 calli under 0.1 mg / L IBA was obtained in hybrid DK236xChi31 (68.8 pcs/100 calli), the smallest one — in PLS61xDK236 (4.3 pcs/100 calli). The highest total regeneration frequency was observed for DK236xChi31 (75.0 %), and the lowest —for PLS61xDK236 (4.4 %) as well.
 In the perspective the obtained outcomes can be used to optimize the biotechnology of maize callusogenesis and plant regeneration in vitro and to increase somaclonal production from long-term cultivated calli for application in breeding process.

CREATION OF IN VITRO COLLECTION AND SEEDS CRYOGENIC BANK OF BERBERIS HETEROPODA

The influence of various seed treatment on germination of eight forms of Berberis heteropoda Schrenk was studied. Laboratory germination (LG) of freshly harvested seeds of this species does not exceed 5%. To stimulate seed germination, the following types of treatment were used – exposure to liquid nitrogen (LN) at -196°C for 20 h, stratification in wet perlite for 2 months at 4°C and stratification with preliminary immersion in LN for 20 h. After LN treatment LG increased by an average of 2.8 times. No germination was observed in samples 1-5 and 8 either before or after exposure to LN, presumably due to the high-water content in larger seeds of these forms. It should be noted that stratification accelerates the energy of seed germination, in addition, the LG after stratification averaged 21.6%, which is 5 times higher than the LG of seeds without additional treatments. The obtained shoots were introduced into in vitro culture on Murashige and Skoog medium with 30 g/l sucrose, 0.8 mg/l 6-benzylaminopurine, 0.02 mg/l indolylbutyric acid, 0.1 mg/l gibberellic acid, 1 mg/l ascorbic acid, 1 mg/l calcium pantothenate, 4 g/l agar, 1.75 g/l Gelrite, pH 5.7. The viability of shoots after testing on specialized medium 523 was 46.4%. Seeds of eight accessions of B. heteropoda were placed in a cryogenic bank at -196°C, 3 accessions were placed for long-term storage at -20°C.  

Микроклональное размножение и особенности адаптации к условиям ex vitro лесных ягодных растений рода Vaccinium

Modern cost-effective propagation methods yield a large amount of high-quality healthy planting material of economically valuable forest berry plants. However, ex vitro adaptation of Vaccinium species and in vitro cultivation of the Kamchatka bilberry remain understudied. The research objective was to study the effect of growth-regulating substances on the organogenesis and adaptation to non-sterile conditions of the lingonberry and the Kamchatka bilberry during clonal micropropagation. The study featured regenerant lingonberries (Vaccinium vitis-idaea L.) of Koralle, Kostromichka, and Kostromskaya Rozovaya cultivars, as well as the Sakhalin and Kuril varieties of the Kamchatka bilberry (Vaccinium praestans Lamb.). A chemical analysis was performed to reveal the following dependencies: the effect of sterilizing agents and sterilization time on the viability of explants, the effect of the nutrient medium and the growth-regulating substances on microshoots and roots, and the effect of the substrate on the survival of plants in non-sterile conditions. The highest survival rate of lingonberry (72%) and bilberry (96%) explants belonged to 0.2% of AgNO3 with 10 min of sterilization time. The maximal values of the total shoot length in vitro were observed at 0.2 mg/L of 2-iP: AN nutrient medium (7.2 cm) for lingonberries and WPM 1/2 nutrient medium (10.5 cm) for bilberries. The longest total root length in vitro for lingonberries was registered when using 2.0 mg/L (5.8 cm) of indoleacetic acid, while for bilberries it was 1.0 mg/L (1.9 cm) of indolylbutyric acid. The maximal survival rate ex vitro belonged to the high-moor peat substrate (89–92%) for lingonberries and a 1:1 mix of peat with sand (91–95%) for bilb erries. Clonal micropropagation with growth regulators (2-iP, indolylbutyric and indoleacetic acids) and peat substrates proved expedient for in vitro cultivation and ex vitro adaptation of the lingonberry and the Kamchatka bilberry. This scheme delivered a large amount of high-quality planting material with high plant survival.

ORGANOGENESE IN VITRO DES FRAGMENTS DE TUBERCULES DE POMME DE TERRE (SOLANUM TUBEROSUM L. VAR. AIDA) CULTIVES DANS LE DEPARTEMENT DES PLATEAUX (REPUBLIQUE DU CONGO)

The production of potatoes rich in starch and essential nutrients does not cover the needs of Congolese consumers. This is due to strong genetic erosion and the absence of high-performance local varieties as well as the unavailability of seeds for new plantations. The seed produced by traditional techniques does not meet the demand of producers. This study aims to produce seed potatoes by the technique of in vitro culture. Tubers disinfected with 10% sodium hypochlorite are kept at 4°C for 20 days to produce microtubers by bud burst others are placed in heat therapy conditions to produce micro-cuttings. Microtubers and microcuttings are taken, then cultured in vitro on Murashige and Skoog medium on the one hand and on Murashige and Skoog medium to which 0.05 mg/ml of indolylbutyric acid is added on the other hand. The cold favors the bud burst of the potato tubers after a week. On vitroplants from microtubers developing on Murashige and Skoog medium to which 0.05 mg / ml of indolylbutyric acid is added, 5 roots, 6 leaves and a stem with a height of 6.05 cm are recorded. On the medium of Murashige and Skoog, we observe 3 roots, 7 leaves and a stem 5.69 cm in height. Regarding microprogation, the results obtained showed that growth hormone plays an important role in the different stages of regeneration compared to the medium without phytohormone.

МІКРОКЛОНАЛЬНЕ РОЗМНОЖЕННЯ ФУНДУКА

For the rapid introduction of large quantities of high-quality hazelnut planting material, it is important to use various modifications of microclonal propagation: classic on gel media, bioreactors with periodic flooding (TiS) and photoautotrophic methods. At the first stage of classical methods there are three main problems: phenol formation; endogenous contamination; selection of trophic and hormonal determinants of in vitro ontogenesis. In cases of infection of mother plants with viruses, viroids, it is mandatory to use meristem explants with a size of not more than 0.3 mm, followed by diagnosis of the effectiveness of recovery. To prevent self-intoxication with phenol-like oxidation products, a set of measures is used to prepare explant donors and modify nutrient media. The most common artificial nutrient media are Driver and Buzzard (DKW) and Nas and Read (NRM). Hazelnuts are sensitive to excess nitrogen and calcium and copper deficiency in artificial nutrient media. On media with a high N content, regenerants have shortened and thickened shoots, often with signs of hypehydration. Inhibition of calcium uptake by nitrogen leads to necrotization of shoot tips and root tips. The incorporation of iron ions into the metabolism of a plant depends on their valence and the form of chelating agents. During the multiplication and induction of rhizogenesis in aseptic conditions, mainly synthetic cytokinin benzylaminopurine and auxin indolylbutyric acid are used. The effectiveness of hormones increases with the use of combinations within groups, in particular cytokinins are benzylaminopurine and kinetin. At the stage of multiplication there is a predominance of cytokinins over auxins, and at the stage of rhizogenesis the number of auxins in the environment is greater than the number of cytokinins. Of the organic components, vitamins B1, B6, C, PP, amino acids and inositol are added to the environment. Ex vitro acclimatization of plants is carried out on peat-pearlite substrates in closed soil or modules of photoautrophic microclonal propagation with increased light intensity and high carbon dioxide content. The photoautotrophic method combines animation, rhizogenesis and acclimatization at the same time. The effectiveness of adaptation to factor-nestatic conditions increases with mycorrhizae by fungi of the genera Glomus, Trichoderma, Tuber in the transfer of plants from aseptic conditions.

Morphogenesis of cell lines of plants Lysimachia nummularia L. at in vitro culture promising for the content of biologically active substances

Lysimachia nummularia L. is a plant belonging to the Primulaceae family, which is particularly valuable as a medicinal raw material used in folk medicine in many countries. It has excellent antibacterial and antioxidant capacity of metabolites. That is why the microclonal propagation of Lysimachia nummularia L. is a relevant issue. The purpose of this paper is to develop approaches to microclonal propagation of L. nummularia. For microclonal reproduction of L. nummularia, the method of activation of isolated tissues and organs already present in the plant meristem and induction of direct regeneration directly by explant tissues was used. To select highly productive cell lines of representatives of the genus Lysimachia, callus culture was obtained by indirect morphogenesis from stem and leaf explants. It was found that the formation of tissues and organs of L. nummularia into in vitro culture depended on the composition of the nutrient medium and the quantitative and qualitative ratio of growth regulators in it. Active proliferation of L. nummularia microshoots into in vitro culture was noted on the variants of Murashige and Skoog, and Driver and Kuniyuki nutrient medium with 6-benzylaminopurine 4.0 mg∙l -1, indolyl butyric acid 0.03 mg∙l -1, gibberellic acid 0.1 mg∙l -1. It was established that for microclonal reproduction, induction, and proliferation of the root system and obtaining regenerating plants of L. nummularia, the most effective is the use of nutrient media according to Murashige and Skoog with the addition of thidiazuron 0.5 mg∙l -1 and 0.25 mg∙l -1 kinetin. The optimal conditions for the induction of callusogenesis and obtaining the culture of cells and callus tissues of L. nummularia and its passage in vitro were selected. It has been shown that the modified nutrient medium of Murashige and Skoog, with 2.4-dichlorophenacetic acid 1.5 mg∙l -1 and indole-3-acetic acid 0.2 mg∙l -1, is optimal for the accumulation of callus tissue biomass of L. nummularia, which ensured the frequency of callusogenesis for the first and second passages up to 98.0 ± 0.2%. 5 cell lines that actively synthesize stilbenoids and the highly productive LN-EE 02/19 cell line, which is capable of synthesizing and accumulating in callus tissues up to 10-12 mg∙g-1 of myricetrin, were selectively isolated. As a result of the analysis, the callus culture cell line LN-EE 02/19 was obtained, which allows obtaining myricetrin in amounts up to 10.0-12.0 mg∙l -1 of raw biomass. The developed protocol can be used both for L. nummularia plants and other representatives of the Primulaceae family

World experience and prospects of hazelnut and almond breeding in Ukraine

Hazelnuts and almonds are promising industrial crops that can differentiate the risks posed by climate change. Domestic almond varieties adapted to local conditions have been created in Ukraine, which require rapid propagation. Creating highly productive and disease-resistant varieties of hazelnuts also requires the reproduction of this crop in signifcant quantities. Of all the methods of vegetative propagation microclonal is the most promising one. Prior to introduction into aseptic culture, mother plants are grown in conditions that minimize endogenous accumulation in tissues of contaminants and phenolic substances. Sodium hypochlorite and Blanidas 300 are used for decontamination of explants. The addition of PPM biocide to the nutrient medium is promising to improve the sterilization of hazelnut explants. The decrease in contamination of almond explants decreases with increasing concentration of Cu, Ag ions in the environment. Mineral nutrients and phytohormones are hte main determinants of in vitro ontogeny. Along with the classic media (DKW, QL, MS, WPM) NRM environment is promising for hazelnuts, and NAM environment – for almonds. Excess in the environment of some elements reduces the assimilation of others, which is manifested visually. Thus, an excess of nitrogen causes symptoms of insufcient absorption of calcium, copper. This process is intensifed as the temperature increases. In addition, regenerating shoots are shortened, thickened and have excessive tissue hydration. Turkish scientists have proposed an original method of creating recipes for almonds and hazelnuts on the quantitative content of elements in the seeds of these plants. Cytokine are the most common phytohormones at the multiplication stage. For both cultures, most researchers prefer benzylaminopurine. Indolylbutyric acid is most often used to induce rhizogenesis. A perlite-based substrate is effective for the regenerants adaptation. It is suggested to inoculate regenerants on bitter almond seedlings as a type of adaptation. A method of photoautrophic microclonal propagation has been developed in Ukraine. These technological methods, which, along with vegetative propagation, allow the adaptation of regenerants in a short time. The efciency is achieved by activating the processes of photosynthesis due to increased carbon dioxide content and light intensity. Key words: hazelnuts, almonds, climate change, microclonal reproduction, determinants, nutrient media, phytohormones.

Preservation in a cryobank of Juglans regia L. accession from several populations in the Sairam-Ugam State National Natural Park

Persian walnut (Juglans regia L.) is one of the most economically important nut crops. In Kazakhstan, the wild-growing population of Juglans regia is registered in the south of the country and is protected on the territory of the Sairam-Ugam State National Natural Park. As a result of expeditions in 2018-2019 136 accessions of Juglans regia were described according to the descriptors of the International Plant Genetic Resources Institute, fruits of 73 accessions of walnuts were collected, which are highly diverse in terms of size, shape and weight of nuts, kernel yield, color and texture of the shell and other parameters. To conserve the biodiversity of Juglans regia, the efficient cryopreservation technique has been developed for isolated embryonic axes. After surface disinfection with 0.1% mercuric chloride solution and rinsing with sterile distilled water, the embryonic axes were desiccated under laminar flow 1 hour to moister content of 12.1%, placed in cryovials, and immersed in a Dewar vessel with liquid nitrogen. In 72.4% of embryonic axes after cryogenic storage, plant recovery was noted on the Driver-Kuniyuki walnut medium with 1 mg/l 6-benzylaminopurine and 0.01 mg/l indolylbutyric acid. A cryogenic collection of 73 Juglans regia accessions has been established. For the first time, the genetic material of Juglans regia, growing in Kazakhstan, has been stored in a cryogenic bank at -196°C. Key words: Juglans regia L., walnut, cryopreservation, cryobank, isolated embryonic axes

Совершенствование методики клонального микроразмножения перспективных сортов Lonicera caerulea L.

The work is devoted to the optimization of clonal micropropagation method of the Lonicera caerulea L. cultivars. The influence of various components of the nutrient medium at all stages of cultivation in vitro has been established. The highest multiplication factor in varieties Diana and Yugana was noted with the addition of glucose at a concentration of 40 g/l and the addition of sucrose at a concentration of 20 g/l. When studying the effect of auxin type on the root formation of honeysuckle varieties, the highest percentage of rooting in Yugana varieties was observed on a nutrient medium with indolylbutyric acid (99 %), and in Gzhelka variety with indoleacetic acid (96 %). It was found that 200 mg/l of iron chelate (Fe(III)-EDDHA) in the composition of the nutrient medium had a positive effect on the percentage of root formation of the variety Diana 81 %. For the Yugana variety (rooting rate 76 %), it is better to use a nutrient medium with the addition of iron chelate Fe(III)-EDTA at a concentration of 73,4 mg/l.

Clonal Micropropagation, Rhizogenesis and Adaptive Capacity of Certain Rose (Rosa L.) Variety Explants

This article presents research on the inclusion of the biotechnological link in rose breeding. Roses are heterozygous plants and give splitting when propagated by seed. Therefore, clonal micropropagation has been included in the research to reproduce and preserve valuable genotypes and new starting material. The aim of this study was to select the type of auxin and its optimal concentrations for better Rhizogenesis of micro-sprouts of selected rose cultivars, and to select the suitable substrate for more effective adaptation of regenerant plants. For the introduction of explants into culture, cultivation and preparation of nutrient media, the traditional methods adopted in tissue culture work were applied. For rooting of well-formed micro-sprouts, liquid nutrient medium with a halved content of macro- and microsalts and sucrose was used. In addition, different concentrations of 3-indolyl butyric acid and β-indolyl-3-acetic acid were added to the medium.Based on the findings of the study, it is recommended to use indolyl butyric acid at concentrations of 0.5 mg/l and 1.0 mg/l for clonal micropropagation of roses, since at these concentrations, the rooting frequency indices by varieties were the highest. When adapting the plant material, it is better to use a substrate consisting of peat, soddy soil and perlite in a ratio of 1:1:1. The best results in the studies were noted in the varieties Alan Titchmarsh, Coral Surprise and Tchaikovski.

Dracocephalum charkeviczii Prob. in vitro

We established sterile clones of a medicinal plant dragonhead Dracocephalum charkeviczii Prob. (Lamiaceae). We used shoots of the plants from natural habitat, notably leaf nodes with axillary bud, as explants, sterilized the explants in 0.1 % Diocide for 4 minutes and cultivated them on agarized nutrient half-strength MS medium with vitamins, 2 % sucrose, 0.4 % agar, 1 mg/l indolylbutyric acid (IBA), or 0.5 or 1.0 mg/l 6-benzyladenine (BA). Medium pH was 4.6 or 5.4. We took hormonefree half-salt MS medium as a control. We observed the maximum propagation coefficient equal to 9.4 in microplants cultivated on the medium with 1 mg/l BA. Application of hormone-free MS medium can be recommended for D. charkeviczii propagation due to its simplicity. The use of the medium protocol with 1 mg/l BA on the first step and with IBA on the second step will allow rapid multiplication and establishment of full-fledged plants of D. charkeviczii for different purposes such as restoration of natural populations, source of valuable metabolites, landscaping, etc.