Skoog Nutrient Medium Research Articles

Изучение влияния препарата Янтарин в составе питательной среды Мурасиге и Скуга на клональное микроразмножение некоторых сортов земляники садовой

The article presents the results of studies on the influence of different variants of modification of Murashige and Skoog (MS) nutrient medium with the help of preparations of growth-regulating action. The researches were carried out on the basis of the educational and production laboratory of agricultural biotechnology of FGBOU VO Kostroma State Agricultural Academy. Regenerant plants of garden strawberry: Asia and Albion were used as research objects. As research subjects were used: growth regulator of cytokinin nature 6-BAP (6-benzinaminopurine), which is standard in plant biotechnology and growth-stimulating preparation - Yantarine, VRK. In the process of research work, a comparative analysis of the effect of different doses of cytokinin 6-BAP and Yantharin preparation on the growth and reproduction of plants of the studied varieties of garden strawberry was carried out. As a result, it was found that at the stage of clonal micropropagation of garden strawberry varieties Asia and Albion, the use of the preparation Yantarine (10.0 mg/L medium) in the nutrient medium together with cytokinin 6-BAP (0.5 mg/L) and 6-BAP (1.0 mg/L) in comparison with the traditional variant 6-BAP (0.5 mg/L medium) - control did not have a stimulating effect on the increase in height and number of micro rosettes formed by each plant regardless of variety. Keywords: CLONAL MICROPROPAGATION, MULTIPLICATION COEFFICIENT, GARDEN EARTH, VARIETY, GROWTH REGULATOR, IN VITRO

Silver nanoparticle-induced in vitro flowering in Dendrocalamus strictus (Roxb.) nees and genetic fidelity assessment of regenerants using molecular markers.

Dendrocalamus strictus (Roxb.) Nees, generally referred to as 'Male bamboo,' is a globally prevalent and highly significant species of bamboo. It is a versatile species and possesses notable industrial significance. However, despite its numerous applications, the production of this plant is insufficient to fulfill the worldwide demand. The challenges that impede the dissemination of D. strictus encompass the unpredictable blooming pattern (30-70 years), low seed production, and limited seed viability. Therefore, tissue culture presents a reliable and effective option for the mass production of standardized planting material. This study investigated the effects of silver nanoparticles (AgNPs) at a concentration of 6.0 mg L- 1 in the Murashige and Skoog (MS) nutrient medium fortified with pre-optimized plant growth regulators (3.0 mg L- 1 6-benzylaminopurine + 0.5 mg L- 1 α-naphthalene acetic acid) on the induction of flowering in a controlled environment in D. strictus. The use of AgNPs in the media induced a maximum of 14 inflorescences per culture vessel, 9 flowers per inflorescence, and improved the performance of the micropropagated plantlets during acclimatization in the greenhouse and field. The ISSR and SCoT amplified polymorphic DNA analysis of the regenerants resulted in the formation of 49 bands (300 to 2000bp size) and 36 scorable bands (350 to 2000bp) respectively. All the PCR amplicons produced by SCoT and ISSR were monomorphic confirming the genetic uniformity of the tissue cultured plants of D. strictus with the mother plant. It can be inferred that the incorporation of AgNPs during the shoot proliferation phase has the potential to stimulate in vitro flowering in D. strictus. This finding could provide valuable insights into innovative strategies for enhancing crop productivity and genetic manipulation for accelerated breeding and agricultural advancement.

Influence of FeSO4.7H2O, Indole-3-Butyric Acid and Different Nutrient Medium on In Vitro Sapling Propagation and Micrografting of Walnut (Juglans Regia L.)

Walnuts are considered a functional food and play a significant role worldwide in people's regular diets. The cultivation and trade of walnut rootstocks and saplings are crucial agricultural activities globally and in Türkiye. Rootstocks are used for grafting and propagating different walnut varieties, and they are essential for tree development, fruit yield, product quantity, and most importantly, their ability to adapt to various ecological conditions and resist diseases and pests. Fruit growing and production of fruit rootstocks of deep-rooted plants in horticulture is given high importance in many countries. However, poor rooting and slow-growing sapling prevent establishing high-yield clonal production. Especially walnut sapling propagation is more complicated and time-consuming than other woody plants. Our research aims to grow high-quality saplings by using biotechnological techniques. This study used modifications to the current in vitro basal medium and evaluated the effects on in vitro rooting and micrografting of walnut species. The optimal medium for shoots induction (2.93±0.90) of the plant was determined as 4 number medium (Murashige and Skoog (MS) nutrient medium with 5.4 µM NAA + 6.9 µM TDZ + 0.6 µM GA3 and solidified with 0.7% agar). In trials to develop sapling, the 10.33 and 6 rooted (2.5cm root length) shoots were obtained from MS nutrient medium with 14.7 and 19.6 µM of IBA and 348.42 µM of FeSO4. 7H2O, and two micrografted plants in MS medium with 19.6 µM of IBA and 348.42 µM of FeSO4.7H2O survived. After three months in the growth chamber, six saplings were ready for transfer to the field. Our findings suggested that FeSO4.7H2O and high dose IBA treatment in MS medium is efficient for in vitro rooting and obtaining in vitro micrografted saplings.

FEATURES OF IN VITRO CULTIVATION OF ASTRAGALUS DASYANTHUS PALL PLANTS

The research is devoted to the study of the peculiarities of in vitro cultivation of the medicinal plant Astragalus dasyanthus Pall, which is listed in the Red Book of Ukraine and has the status of an endangered species. The cultivation of woolly-flowered astragalus in Ukraine is currently difficult due to the inaccessibility of the raw material base in the temporarily occupied southern part of Ukraine, as well as due to the low seed productivity of the crop. The use of in vitro plant tissue cultivation methods to restore natural communities of A. dasyanthus and expand possible sources of raw materials for the synthesis of medicinal substances is promising. In this work, we investigated the conditions for obtaining aseptic Astragalus dasyanthus plants from seeds and evaluated the effect of preliminary mechanical scarification on its germination energy, taking into account the percentage of sterile seedlings obtained. In addition, the efficiency of microclonal propagation of A. dasyanthus was determined depending on the composition of the culture medium, in particular the concentration of macronutrients and the presence and concentration of growth regulators. It has been shown that aseptic plants of Astragalus dasyanthus can be obtained by surface sterilization of seeds that have been previously subjected to mechanical scarification or by germination on Murashige and Skoog nutrient medium with the addition of 0.1 mg/l benzylaminopurine. It was determined that the type and concentration of cytokinin affects the efficiency of microclonal propagation of A. dasyanthus plants, while halving the content of macronutrients in the nutrient medium does not significantly affect this item. It was found that the addition of benzylaminopurine to the culture medium stimulates the formation of more additional shoots of A. dasyanthus in vitro compared to the culture medium containing kinetin. Thus, Murashige and Skoog medium with the addition of 0.5 mg/l of benzylaminopurine is optimal for in vitro cultivation of woolly-flowered Astragalus plants.

Propagation of Canadian rose ‘John Franklin’ in vitro culture

Many researchers are interested in optimizing the clonal micropropagation of ornamental plants. It is known that the varietal specificity of the studied crops makes it necessary to select and optimize the composition of the nutrient medium for each stage of propagation in vitro. The purpose of our research was to identify the effect of the composition and concentration of nutrient hormones on multiple shoot formation, root development in regenerating plants of the rose 'John Franklin' in vitro. A technology has been developed for obtaining a large amount of self-rooted planting material of a winter-hardy decorative rose variety in vitro culture. Clonal micropropagation was performed using vegetative buds as explants. The multiple of shoots is achieved by activating the axillary meristems of the shoots. It has been shown that the efficiency of micropropagation of the 'John Franklin' rose increases on the Murashige and Skoog nutrient medium containing BAP at a concentration of 1.0 mg/l and IAA – 0.5 mg/l. This medium provides high quality regeneration of micro-shoots with a propagation coefficient equal to 5.2. The optimal modification of the nutrient medium at the rooting stage was revealed, and the expediency of using IBA with a concentration of 0.5 mg/l as auxin was noted. The selected conditions allow you to get a larger number of roots on the shoot – 5 pcs. and their greatest length is 51.5 mm.

Размножение in vitro перспективных сортов картофеля для возделывания в условиях Республики Коми

The paper deals with the results of clonal micropropagation of promising potato varieties for cultivation in the conditions of the Komi Republic. The study material is three potato varieties (Zyryanets, Pechorsky, Vychegodsky), characterized by the high resistance to late blight and abiotic stress factors in the conditions of the Komi Republic. The primary explants were weakly germinated eyeholes, as well as cuttings and shoot tips isolated from 14- and 30-day-old potato plants which underwent surface sterilization and then were cultivated in the Murashige–Skoog nutrient medium containing various hormones. The Pechorsky variety was identified to have the high growth index and specific growth rate of shoots in vitro. This variety can be included in studies on cell and genetic engineering aimed at creating potato forms with the complex resistance.

Antioxidant and Antitumor Potential of Micropropagated Balkan Endemic Sideritis scardica Griseb

Sideritis scardica Griseb. is a critically endangered Balkan endemic species, known for its antioxidant, neuroprotective and anti-inflammatory properties. The aim of the present study was to detail an efficient protocol for the micropropagation of S. scardica. In vitro cultures were initiated from the shoot tips of 40 days-old in vivo seedlings and the effects of different plant growth regulator treatments were examined. A Murashige and Skoog nutrient medium (MS) containing 1 mg/L zeatin and 0.1 mg/L indole-3-acetic acid (IAA) proved to be the most efficient for shoot multiplication as it produced quality, vigorous shoots with a mean number of six shoots per explant. For the first time, the antioxidant and antitumor activities of extracts from in vitro-obtained plants were evaluated. In vitro cultivated plants grown in the field revealed a higher total polyphenol content (3929.1 ± 112.2 mg GAE/100 g vs. 3563.5 ± 52.8 mg GAE/100 g) and higher ORAC antioxidant activity (1211.6 ± 27.3 µmol TE/g vs. 939.9 ± 52.4 µmol TE/g) than in situ cultivated plants. A comparison of the antitumor activities of extracts from in vitro propagated shoots, field-grown in vitro-obtained plants and in situ plants on HeLa (cervical adenocarcinoma), HT-29 (colorectal adenocarcinoma) and MCF-7 (breast cancer) human cancer cell lines showed that in vitro propagated shoots had a significant concentration-dependent cytotoxic effect on the cervical adenocarcinoma cell line HeLa, while the field-grown in vitro-obtained and in situ-collected samples induced the highest reduction in the viability of the mammary carcinoma cell line MCF-7. In both cases, the cells of the control non-tumor cell line, BALB/3T3, were significantly less affected. The results showed that the in vitro multiplication protocol ensured the obtainment of numerous plants with antioxidant and antitumor potential.

IMPROVEMENT OF THE CHRYSANTHEMUM × KOREANUM HORT. CLONAL MICROPROPAGATION PROCESSES USING BACILLUS MEGATERIUM ONU500

The aim of this study was to improve the Chrysanthemum × Koreanum micropropagation processes using Bacillus megaterium ONU500 at the post-aseptic adaptation stage. Methods. At the initial stages of clonal micropropagation, two surface sterilization protocols with different fungicidal preparations were tested. The influence of the agar concentration in the medium on the development of explants was determined. At the ex vitro adaptation stage, the roots of the microclones were inoculated with bacteria of the strain B. megaterium ONU500. Results. The most effective protocol for plant material surface disinfection, which included quinozol and ethanol, was established. It was determined that the use of Murashige and Skoog (MS) nutrient medium supplemented with 0.4% agar at the stage of in vitro culture establishment increased the survival of chrysanthemum initial explants by 9.3% and accelerated proliferation by 1 day. It was found that inoculation of the chrysanthemum microclone rhizosphere with bacteria had a positive effect on the growth of adapted plantlets, increasing their survival rate by 20.4–22.2%, shoot height by 1.8–1.9 cm, and average number of nodes by 2.0–2.2 nodes. Conclusion. The effectiveness of the use of fungicides for the treatment of chrysanthemum plant material during in vitro culture establishment and the influence of the consistency of the nutrient medium on the development of explants were determined, which is new and relevant for increasing the efficiency of clonal micropropagation of this plant. Also, experiments on the inoculation of chrysanthemum microclones at the stage of ex vitro adaptation were carried out for the first time, and the positive effect of such inoculation on the morphometric characteristics of seedlings and their survival rate was shown. When establishing chrysanthemum explants in vitro, it is recommended to use the fungicide quinozol for surface sterilization, and carry out cultivation in a semi-liquid nutrient medium with 0.4% agar. At the postaseptic adaptation stage of microclones, it is advisable to inoculate the roots with bacteria of the strain B. megaterium ONU500 at concentrations of 108–109 CFU/ml.

Influence of nutrient media on the physiological parameters of grape microclones

Successful rooting of grape microclones in uncontrolled environmental conditions (in vivo) depends on the level of resistance, which is formed at the stage of their passage and growth in vitro. An important role in this case is played by indicators of the water regime of the vegetative mass. The purpose of this study was to get acquainted with the results of determining the water-holding capacity, the intensity of transpiration of grape microclones in vitro and to establish the share of their influence on the adaptation potential in vivo. Biotechnological, laboratory, vegetation, and mathematical and statistical research methods were used in this study. The obtained results showed that to optimize the physiological processes in the tissues of the leaves and shoots of grape microclones, to increase their viability under in vivo conditions, it is advisable to cultivate them in vitro on structured nutrient media (MS + agroperlite, MS + vermiculite, MS + agroperlite + vermiculite) with the content of phytohormones IAA – 0.2 mg/l, and 6-BAP – 0.3 mg/l. Structured nutrient media contributed to an increase in water retention capacity and a decrease in the intensity of transpiration of tissues of leaves and shoots of microclones of both technical and rootstock varieties. During 60 minutes of research, from 0.006 g to 0.034 g of water evaporated from microclones of technical varieties, from 0.003 g to 0.053 g from microclones of rootstock varieties, respectively. Transpiration intensity (after 10 min) decreased by 1.7-1.8 times. On the control Murashige and Skoog nutrient medium, plants evaporated a larger amount of water during the corresponding period of time: from 0.006 g to 0.079 g (technical varieties) and from 0.008 g to 0.086 g (rootstock varieties); the intensity of transpiration was higher. After cultivation of grape microclones on structured nutrient media, they were characterized by a higher content of dry matter in the tissues of leaves and shoots (14.6-15.0%) and better survival rates in vivo (76.3-98.5%, at 58.5-65.2% in control). The reliability of the results obtained is confirmed by the results of multivariate analysis of variance. The obtained results expand the understanding of dynamic changes in the indicators of the water regime of the vegetative mass of grape microclones in vitro, their influence on plant survival under in vivo conditions

Preliminary Study: Micropropagation Using Five Types of Chelated Iron and the Subsequent Acclimation of Blue Honeysuckle (Lonicera caerulea var. kamtschatica Sevast.)

Blue honeysuckle (Lonicera caerulea var. kamtschatica Sevast.) is a valuable berry crop with a unique biochemical composition. It is rich in vitamins, minerals, and biologically active substances. Different species and cultivars of honeysuckle require different cultivation conditions in the field of accelerated reproduction in vitro. Taking into account the high clonal replication potential of the plant, we conducted research on the chelated-iron form’s influence on the micropropagation productivity of the blue honeysuckle “Lulia” cultivar at the multiplication, rooting, and subsequent acclimatization stages of microplants. In a preliminary study, five types of iron chelates were tested with carboxyl- and phosphorus-containing ligands: Fe(III)-EDTA, Fe(III)-DTPA, Fe(III)-EDDHA, Fe(III)-HEDP, and Fe(II)-HEDP. Each type of iron chelate was applied at four concentrations: standard, decreased by 2 times, and increased by 1.5 times and 2 times in the basic Murashige and Skoog (MS) nutrient medium. It was found that the blue honeysuckle “Lulia” had a selectivity to the type of iron chelate that was used. The nutrient-medium modifications with iron chelates, which caused the plant response, contributed to a significant improvement in the plant’s physiological status and increased its survival rate during the microplant’s acclimation to the nonsterile conditions stage. At the rooting stage, an increase in the rooting rate of up to 100% (Fe (III)-EDDHA), an increase in the number of shoots by 1.5–2 times, and an increase in the number of roots by 1.4–1.9 times were observed. The positive effect of the iron’s chelated forms was also observed at the acclimation stage. According to the results of the research, the most suitable iron forms for clonal micropropagation of blue honeysuckle were carboxyl-containing Fe(III)-EDDHA and phosphonate-containing Fe(II)-HEDP. Moreover, the effectiveness of both complexonates was confirmed in a wide concentration range: Fe(III)-EDDHA from (×1.0) to (×2.0), and Fe(II)-HEDP from (×0.5) up to (×1.5).

Induction of garlic (<i>Allium sativum</i> L.) inflorescence callusogenesis <i>in vitro</i>

In order to expand the genetic diversity of winter garlic (Allium sativum L.), the possibility of using in vitro technology to obtain somaclonal variants of a callus culture was studied. The results of the study of the efficiency of inflorescences callusogenesis of winter garlic cultivar ladiator depending on the explant age (7, 14 and 21 days from the moment when inflorescences emerge from the leaf axils) and the combination of the growth regulators (2,4-dichlorophenoxyacetic acid in concentrations 0,25; 0,5; 1,0; 2,0 mg/l and kinetin in concentrations 0,25; 0,5; 1,0 mg/l) are presented. It was found that stepwise sterilization of the material, based on sequential treatment with sterilizing solutions, allows obtaining 92-98% sterile explants. The first signs of the beginning of callusogenesis were observed in early garlic inflorescences on the 35th day of cultivation, and in the inflorescences aged 21 days - on the 50th day. Callus formation from the base of inflorescences was recorded when 0.5; 1.0 and 2.0 mg/L of 2,4-dichlorophenoxyacetic acid and kinetin were added to the nutrient medium at the concentrations studied. The most intensive callus formation was detected in 7-day-old explants on the Murashige - Skoog nutrient medium enriched with 2.0 mg/L 2,4-dichlorophenoxyacetic acid and 0.5 and 1.0 mg/L kinetin. The proportion of explants with callus was 94% and 96%, respectively. Callus formation from the base of inflorescences at 14 and 21 days of age using these nutrient media options was observed in 44 and 22% of explants, respectively.

Selection for Sustainable Preservation through In Vitro Propagation of Mature Pyrus spinosa Genotypes Rich in Total Phenolics and Antioxidants

Pyrus spinosa Forssk. (almond-leaved pear) is a wild-growing native tree of the hilly countryside of Greece, which has recently gained interest for its valuable endogenous substances. In the present work, the determination of the total phenol content (TPC) and antioxidant capacity (AC) was carried out during 2021 and 2022 in leaves from 32 genotypes of mature wild-growing trees at four different locations of Central Macedonia District, Greece: Agia Anastasia, Chalkidiki, Lagadas and Pieria. The measurements taken from the leaf samples of all genotypes revealed a strong positive linear correlation between TPC and AC (R2 = 0.772). Two genotypes from Agia Anastasia, coded as AA2 and AA3, were comparatively the richest in TPC (44.86 for AA2 and 46.32 mg GAE/g fresh weight for AA3) and AC (70.31 and 71.21 μg AAE/g fresh weight for the same genotypes). For these two genotypes of high TPC and AC, an efficient micropropagation protocol was developed to preserve and multiply this valuable germplasm. Newly emerged shoot tips were excised from winter shoots and, after disinfestation, they were established on a modified Murashige and Skoog (MS) nutrient medium with 5 μΜ of 6-Benzyloaminopurine (BAP). For shoot multiplication, explants from clean cultures were transferred to Pear Medium 1 with 5 μΜ of BAP. The effect of the nutrient medium was tested by using five different nutrient media (modified MS, Pear Medium 1, Pear Medium 2, DKW and WPM) supplemented with 5 μΜ of BAP. Pear Medium 1 was the most effective in shoot formation. Among the four different BAP concentrations (0, 5, 10 and 20 μΜ) used in Pear Medium 1, 5 μΜ of BAP resulted in the production of the significantly highest number of shoots (22.7 shoots per explant, 2.4 cm long). The exposure of cultures to 10 μmol·m−2·s−1 for one week under light irradiance followed by 35 μmol·m−2·s−1 for four weeks increased both the number and length of the shoots produced. A 20-s dip of the shoot bases into 49.0 μΜ of Indole-3-butyric acid (IBA) in EtOH equally affected rooting in a modified rooting MS (79%) and Pear Medium 1 (80%). The root system developed better in the modified rooting MS medium (mean number of roots of 2.7 with a length of 3.9 cm). The acclimatization of the plantlets was successfully carried out (high survival rates five weeks after their establishment) on a perlite substrate under controlled environmental conditions.

The Effect of Some Heavy Metals on the Growth of Garlic under In Vitro Conditions

Garlic (Allium sativum L.) is an important plant species because of its nutritional and medical value. One of the important advantages of plant tissue culture is in vitro selection. The tolerance of garlic to some heavy metals under in vitro conditions was studied. In vitro experiments were carried out in Murashige and Skoog (MS) nutrient medium supplemented with different doses of cadmium (Cd), cobalt (Co), nickel (Ni), aluminum (Al), copper (Cu), chromium (Cr), and lead (Pb) at 100 µm, 200 µm, 300 µm, 400 µm, and 500 µm. During experiments, plant length, number of leaves, leaf length, root number, and root length parameters were considered. In the findings, there was a decrease in both leaf and root development as doses of heavy metals increased. In addition, it has been found that the decrease in plant development at doses of 400 µm and 500 µm was higher compared with other doses. There was no root formation at high doses of Cd, Co, Ni, and Cu (400 µm and 500 µm). As a recommendation, further research should be conducted under in vivo conditions to evaluate the level of tolerance to heavy metals in garlic.

IN VITRO PRODUCTION OF ACTIVE COMPOUNDS IN THE CALLUS OF THE CHIA PLANT (SALVIA HISPANICA)

The latest research aimed to study the effects of plant growth regulators on inducing callus from the chia plant (Salvia hispanica) and stimulating it to produce some medicinal compounds in vitro. The study transpired at the Plant Tissue Culture Laboratory, College of Agriculture, University of Anbar, Iraq. The use of Murashige and Skoog (MS) nutrient medium containing 2,4-D Auxin (dichlorophenoxyacetic acid) had concentrations of 1.0, 2.0, and 3.0 mg L-1 and cytokinin benzyl adenine (BA) with concentrations of 0.0, 0.5, and 1.0 mg L-1. The use of their interactions continued to induce a perpetuation of callus. In another experiment, the salicylic acid at concentrations of 0, 10, 20, and 30 mg L-1 stimulated the callus to produce medicinal compounds. The results showed that for induction of callus and its sustainability in the leaves of the chia plant, the best concentration revealed 2,4-D at a rate of 3 mg L-1, which achieved the highest fresh and dry weight and dry matter percentage with values of 0.5151 g, 0.0723 g, and 13.531%, respectively. The results also showed that the addition of salicylic acid at the concentration of 10 mg L-1 to the nutrient media stimulated the formation of the active compounds.

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.

Methodological aspects of increasing the intensity of callusogenesis and organogenesis of Linum usitatissimum L. in vitro

Aim. Improving methods for increasing the efficiency of obtaining callus cultures and somaclones of flax (Linum usitatissimum L.) in vitro. Methods. Hypocotyl segments were cultured on Murashige and Skoog nutrient medium supplemented with sucrose (30 g/l) and phytohormones at various concentrations. Other conditions: photoperiod 16 hours, illuminance 2500 lx, relative humidity 60–80%, air temperature 22–24°C. Results. The ability to form callus and somatic embryogenesis of flax depends on the phytohormonal composition of the nutrient medium, the size of the explants and the distance between them. Conclusions. For intensive callus formation and somatic embryogenesis in vitro, the optimal concentrations of BAP (mg/l) can be expressed as 1.0 ≤ BAP ≤ 1.75; the optimal concentrations of BAP for the medium supplemented with NAA (0.05 mg/l) 0.5 ≤ BAP ≤ 2.0; the optimal concentration of NAA for the medium supplemented with BAP (1.0 mg/l) 0.025 ≤ NAA ≤ 0.150; and the optimal concentrations of IAA for the medium supplemented with BAP (1.0 mg/l) 0.05 ≤ IAA ≤ 0.50. Addition of 0.5 mg/l GA3 to the medium with NAA and BAP is effective. It is optimal to use hypocotyl explants 3–6 mm long and place them at a distance of 1.5–2.5 cm from each other. Organogenicity of callus is significantly reduced in the process of subculturing.

Sustainable Exploitation of Greek Rosmarinus officinalis L. Populations for Ornamental Use through Propagation by Shoot Cuttings and In Vitro Cultures

Rosmarinus officinalis L. belongs to the Lamiaceae family and is cultivated worldwide due to its diverse uses as an ornamental evergreen shrub in gardening, as well as a food seasoning and a natural medicine. The present research was conducted to study the morphological traits of seven wild-grown rosemary populations in Greece, as well as the propagation of two selected populations with the most desirable ornamental characteristics, by the use of shoot cuttings and in vitro cultures. From the study of the morphological traits of the seven populations, those with interesting features were grown in the areas of Amaliada and Piperia and, thus, were chosen for studying their propagation. Rooting of the shoot cuttings was carried out during the four seasons of the year, using potassium salt Indole-3-butyric acid (K-IBA) at concentrations of 0.5 and 1 g·L−1, in various substrates under the intermittent mist or fog system. It was found that the shoot cuttings rooted easily in all four seasons, but there were better results for the population of ‘Piperia’ in autumn, with 1 g·L−1 K-IBA (80%), and ‘Amaliada’ in spring, with 0.5 g·L−1 K-IBA (82.5%), while higher rooting percentages were achieved in the fog system, on a substrate consisting of perlite and peat, in a ratio of 2:1 (85%). For the in vitro cultures, shoot tips excised from the two selected populations were successfully disinfested by pre-soaking in an antioxidant solution and then, by sterilizing them in 0.6% (w/v) NaOCl, followed by transferring them onto a Murashige and Skoog (MS) nutrient medium. ‘Amaliada’ cultures proved to be the most productive population (2.1 shoots per explant), with the highest shoot formation frequency (91.6%), when cultivated on the MS nutrient medium without plant growth regulators. For ‘Piperia’ cultures, the highest shoot formation frequency (66.6%) was achieved on the MS nutrient medium supplemented with 0.25 or 0.5 mg·L−1 6-Benzyloaminopurine (BAP) and 0.1 mg·L−1 Indole-3-acetic acid (IAA). Spontaneous root formation frequency was noticed on the MS nutrient medium, containing 0.5 mg·L−1 BAP and 0.1 mg·L−1 IAA, for both ‘Amaliada’ and ‘Piperia’ cultures (50% and 41.6%, respectively) in a single stage, with root lengths of 7.1 and 5.3 cm, respectively. Rosemary plantlets, with roots formed in vitro after transplanting them in soilless substrate, were acclimatized adequately in the greenhouse environment (~70%).

Innovative technologies for cloned plants adaptation

Chrysanthemum plants are popular with amateur gardeners and breeders. It is widely used in landscaping cities, parks, and also for bouquets. Plants are notable for longer flowering and a bright color of flowers. Extracts of chrysanthemum plants contain various biologically active substances with bactericidal properties and are used in pharmacology and dentistry. The objective is to develop an effective technology for rapid in vitro reproduction and adaptation of chrysanthemum microclones. The objects of the study were chrysanthemum plants of three varieties: Bacardi, Korean Dawn, and Snow White. Semifloscules and flower buds were used as primary explants. The explants were cultured on a Murashige and Skoog (MS) nutrient medium containing mineral salts, as well as various growth regulators: 1) 1 mg L-1 BAP + 0.5 mg L-1 IAA; 2) 15% Aminoven solution 3 mL L-1 ; 3) Stimul 1 mL L-1 ; and 4) a hormone-free medium. Microshoots were adapted using an aeroponic system. Unrooted microshoots were used for adaptation. The GrowPlant X-Stream 120 propagator was used as equipment for adaptation. The frequency of adventious budding depended on the hormonal composition of the nutrient medium. Aminoven in the nutrient medium led to the induction of the formation of adventitious buds on average in 85% of cases. The same indicator averaged 72% on the Stimul-containing medium. In a BAP-containing medium, the considered indicator was the same with the control variant (hormone-free medium). The Snow White explants had the highest regenerative activity, Bacardi was second, and Korean Dawn was third. Studies have shown that the efficiency of adaptation of unrooted microshoots under aeroponic conditions is significantly influenced by the composition of the nutrient medium during micropropagation. Thus, preliminary cultivation of microshoots in vitro on Stimul and Aminoven-containing media led to the rapid formation of the root system and active growth of the aerial part. In these variants, the shoot growth index (I) was 0.65-0.71, and specific rate (µ) was 0.89-1.1 mm day-1.

Effect of 1-naphthylacetic and indol-3-acetics acid on the intensity of callusogenesis and organogenesis of Linum usitatissimum L. in vitro

Aim. Investigate the effect of auxins of exogenous origin in nutrient medium in vitro on the germination and organogenesis intensity in Linum usitatissimum L. convar. elongatum (‘Hlinum’ variety) at the constant concentration of 6-benzylaminopurine (BAP). Methods. Hypocotyl segments were cultured on Murashige and Skoog nutrient medium supplemented with sucrose (30 g/l) and phytohormones at various concentrations. Other conditions: photoperiod 16 hours, relative humidity 60–80%, air temperature 22–24°C. Results. Common flax has a great capacity to form callus and shoots under the effect of the following factors: 1) only auxins, 2) only cytokines, 3) combinations of auxins and cytokines. Somatic embryogenesis is also possible on a nonhormonal nutrient medium. Conclusions. For somatic embryogenesis in vitro, the optimal concentrations of BAP can be expressed as 1.0 ≤ BAP ≤ 1.75, the optimal concentrations of BAP for the medium supplemented with 1-naphthylacetic (NAA, 0.05 mg/l) 0.5 ≤ BAP ≤ 2.0, the optimal concentration of NAA for the medium supplemented with BAP (1.0 mg/l) 0.025 ≤ NAA ≤ 0.150, and the optimal concentrations of indol-3-acetics acid (IAA) for the medium supplemented with BAP (1.0 mg/l) 0.05 ≤ IAA ≤ 0.50. Keywords: Linum usitatissimum L., in vitro, phytohormones, callus, organogenesis.

Bazı Biber (Capsicum annuum L.) Genotiplerinin Anter Kültürü Performanslarının Belirlenmesi

Bu çalışmada, farklı tiplerde 23 biber genotipinde anter kültürü performansı belirlenmiştir. Çiçek tomurcuklarının dezenfeksiyonunda, tomurcuklar öncelikle % 70’lik etil alkolde 30 saniye bekletilmiş ve sonrasında 10 dakika % 15’lik sodyum hipoklorit çözeltisinde tutulmuşlardır. Anterler ilk aşamada; 30 g/L sakkaroz, 2.5 g/L aktif kömür, 15 mg/L gümüş nitrat (AgNO3), 4 mg/L 1-Naphthaleneacetic acid (NAA), 0.5 mg/L 6-Benzylaminopurine (BAP) ve 6.5 g/L agar içeren Murashige and Skoog (MS) besin ortamında kültüre alınmışlardır. Sonrasında anterler, 30 g/L sakkaroz ve 6.5 g/L agar içeren ve büyümeyi düzenleyici içermeyen MS besin ortamına aktarılmışlardır. Elde edilen embriyolarda, aynı besin ortamına alınmışlardır (ikinci ortama). Anterler, ön uygulama olarak, +35ºC’de ve karanlık koşullarda 2 gün süresince bekletilmişlerdir. Çalışma sonuçları değerlendirildiğinde, genotiplere göre 100 anter için ortalama embriyo sayıları 0.83 ile 44.44 arasında değişmiştir. En yüksek oran, FT-509 genotipinde gözlemlenmiş ve bu genotipi FT-508 (23.61 embriyo/100 anter), FT-1181 (23.37 embriyo/100 anter) ve FT-905 (22.89 embriyo/100 anter) genotipleri takip etmiştir. En düşük performansı, 0.83 adet embriyo/100 anter ile FT-1178 genotipi göstermiştir. Oluşan embriyoların tamamı, bitkiye dönüşebilmiştir. Bu nedenle, 100 anter için ortalama bitki oluşturan embriyo sayısı değerleri, 100 anter için ortalama embriyo sayıları ile aynı olmuştur. En yüksek ortalama bitki sayısı 20 adet ile FT-508 no’lu genotipte gözlemlenmiş ve FT-509 (17.5 bitki), FT-1181 (16 bitki), FT-905 (12 bitki), FT-263 (10.5 bitki) ile FT-507 (8.25 bitki) genotipleri tarafından takip edilmiştir. En düşük bitki sayısı ise 0.5 adet ile FT-1178 no’lu genotipte kaydedilmiştir.