Plant Micropropagation Research Articles

Acidification of the culture medium as a strategy to control endophytic contaminations in Prunus spp. rootstocks cultured in GreenTray TIS bioreactor

Overgrowth of endophytes in some in vitro cultures may disrupt the normal shoot tip growth and proliferation, being necessary to obtain endophytes-free cultures to achieve a normal plant micropropagation process. To remove these contaminations from the culture medium, antibiotics are commonly added to the culture medium. However, its use in plant production must be urgently reduced because of the current restrictions imposed by the European Union. For that purpose, the effect of acidic low (pH 3) and neutral (pH 7) pH was tested in the GreenTray® TIS bioreactor as an alternative to control endophytes growth without affecting the micropropagation of the Prunus rootstock RP-20 explants. The results demonstrated that culture at pH 3 did not affect the number of shoots, shoot FW, shoot length and the amount of chlorophyll pigments, but significantly reduced endophytes population. The identification also revealed that Roseomonas mucosa, Microbacterium oxydans, Bacillus subtilis and Luteibacter yeojuensis were the bacterial isolates responsible of those contaminations. These results might suppose a real breakthrough in the in vitro tissue culture field, although more research is required to meet the pH requirements for the different plant species and other endophytic microorganisms.

Studies on Phytochemical, Antimicrobial activity and Micro propagation of Medicinal plants from Eastern Ghats of Andhra Pradesh

The aim of the present study was to investigate the presence of phyto chemical, antimicrobial activities and micro propagation of the selected medicinal plants i.e. Rauwolfia serpentina, Adhatoda vasica and Alstonia scholaris. These endemic plants belong to Apocynaceae and were collected from higher altitudes of Eastern Ghats, Lambasingi forest region, Andhra Pradesh India. To determine the total phenolic and flavonoid contents, Soxhlet apparatus was used for this study. Solvents used were in this study are water, ethyl acetate, chloroform and methanol. Among them the solvent methanolic extract of Rauwolfia serpentina (57.15±1.2) and Alstonia scholaris (55.06±0.7) showed high content of saponins. The test microrganism which were studied against the efficacy of selected medicinal plant extracts were, two bacterial strians i.e., Streptococcus pyogenes, Pseudomonas aeruginosa and two fungal strains Aspergillus niger and Vibrio cholera. The antimicrobial activity was proved that the methanol extracts was found to be maximum antimicrobial growth inhibition. The simple and effective protocol was developed to propagate the Rauwolfia plant from nodal explants. Maximum no of 95% plantlets regenerated successfully. These propagated plantlets were hardened to survive in vivo conditions.

Growth of Micropropagated Solanum tuberosum L. Plantlets under Artificial Solar Spectrum and Different Mono- and Polychromatic LED Lights

In agriculture, LED light sources have increasingly replaced the standard luminescent lamps and have acquired an important role in plant micropropagation. We studied the effect of different light sources such as narrow-band LEDs (bright blue, blue, green, yellow, deep red, and red) and wide-band LEDs (cold white, white, warm white, full spectrum, and an artificial solar spectrum sun box constructed by us) on development of potato plantlets in vitro. White luminescent lamps were used as a control. The light intensity of 49 µmol · m−2 · s−1 was provided in all light treatments. We showed that the long-wave narrow-band light treatments were inapplicable for potato micropropagation, because plantlets were weak with small leaves, inhibited roots, and significantly elongated stems. Blue lights provided growth of shortened plantlets with large leaves, well-growing roots, and abundant green mass. The chlorophyll content was lower under blue and bright blue light and was at the same level in the remained treatments. Significant differences in the stomatal apparatus development were observed depending on the light source. These differences were not always reflected in the plantlet phenotype: e.g., plantlets under blue and bright blue lights showed no differences in any characteristics except stomatal density and size of stomatal guard cells. We found no significant effect of blue light portion in the white lights and full spectrum on plantlet growth. An artificial solar spectrum sun box was the most suitable for potato micropropagation, because it supported the development of plantlets with good fitness, uniform internodes length, abundant roots and green mass accumulation.

The efficiency of interaction between cytokines and Auxins in Micropropagation of Chrysanthemum plant (Chrysanthemum indicum L.)

This study was conducted in the Tissue Culture Laboratory at the College of Science / Tikrit University, with the aim of micropropagation of the Chrysanthemum plant and determining the optimal concentration of the growth regulators used, In this study, a single nodes cutting from the shoots of Chrysanthemum indicum L. was used and the growth regulator (kinetin) Kin with concentrations (0.0-4.0) mg.L−1 time alone and again with the interaction (Indol-3-butyric acid) IBA at concentrations (0.0-0.6 ) mg.L−1, The results showed that the interaction between the concentrations of Kin and IBA had a higher significant effect than the use of Kin alone, Where the average of vegetative growth of Chrysanthemum plant increased as the average number of shoots, shoot length and number of leaves amounted to 7.2 shoot.plant−1, 5.27 cm and 17.0 leaf.plant−1 when added 4 mg.L−1 of Kin with 0.6 mg.L−1 of IBA to Murashige and skoog medium (MS) after being 3.3 shoot.explant−1, 3.81 cm and 5.3 leaf.shoot−1 respectively when 4 mg L-1 of Kin has added alone to MS medium, As for the rooting experiments, in which the average MS was used with the full strength of the salts and half the strength of the salts containing different concentrations of IBA growth regulator (0.0-2.0) mg.L−1 for rooting the shoots resulting from the multiplication experiments. The MS medium half the salt strength excelled and giving the highest average of rooting percentage, average number of roots and average root length of 90%, 12.5 root.explant−1 and 3.5 cm respectively when adding 1 mg.L −1 IBA to the medium.

Potential Role and Utilization of Plant Growth Promoting Microbes in Plant Tissue Culture.

Plant growth promoting microbes (PGPMs) play major roles in diverse ecosystems, including atmospheric nitrogen fixation, water uptake, solubilization, and transport of minerals from the soil to the plant. Different PGPMs are proposed as biofertilizers, biostimulants, and/or biocontrol agents to improve plant growth and productivity and thereby to contribute to agricultural sustainability and food security. However, little information exists regarding the use of PGPMs in micropropagation such as the in vitro plant tissue culture. This review presents an overview of the importance of PGPMs and their potential application in plant micropropagation. Our analysis, based on published articles, reveals that the process of in vitro classical tissue culture techniques, under strictly aseptic conditions, deserves to be reviewed to allow vitroplants to benefit from the positive effect of PGPMs. Furthermore, exploiting the potential benefits of PGPMs will lead to lessen the cost production of vitroplants during micropropagation process and will make the technique of plant tissue culture more efficient. The last part of the review will indicate where research is needed in the future.

Using Micropropagation to Develop Medicinal Plants into Crops.

Medicinal plants are still the major source of therapies for several illnesses and only part of the herbal products originates from cultivated biomass. Wild harvests represent the major supply for therapies, and such practices threaten species diversity as well as the quality and safety of the final products. This work intends to show the relevance of developing medicinal plants into crops and the use of micropropagation as technique to mass produce high-demand biomass, thus solving the supply issues of therapeutic natural substances. Herein, the review includes examples of in vitro procedures and their role in the crop development of pharmaceuticals, phytomedicinals, and functional foods. Additionally, it describes the production of high-yielding genotypes, uniform clones from highly heterozygous plants, and the identification of elite phenotypes using bioassays as a selection tool. Finally, we explore the significance of micropropagation techniques for the following: a) pharmaceutical crops for production of small therapeutic molecules (STM), b) phytomedicinal crops for production of standardized therapeutic natural products, and c) the micropropagation of plants for the production of large therapeutic molecules (LTM) including fructooligosaccharides classified as prebiotic and functional food crops.

Biofertlizer Lumbrical improves the growth and ex vitro acclimatization of micropropagated pear plants

In vitro micropropagation of plants is highly useful for obtaining large quantities of planting material with valuable economic qualities. However, plantlets grow in vitro in a specific environment and the adaptation after the transfer to ex vitro conditions is difficult. Therefore, the acclimatization is a key step, which mostly determines the success of micropropagation. The aim of this investigation was to study the effect of the biofertlizer Lumbrical on ex vitro acclimatization of micropropagated pear rootstock OHF 333 (Pyrus communis L.). Micropropagated and rooted plantlets were potted in peat and perlite (2:1) mixture with or without Lumbrical. They were grown in a growth chamber at a temperature of 22±2 °C and photoperiod of 16/8 hours supplied by cool-white fluorescent lamps (150 µmol m-2 s-1 Photosynthetic Photon Flux Density, PPFD). The plants were covered with transparent foil to maintain the high humidity, and ten days later, the humidity was gradually decreased. Biometric parameters, anatomic-morphological analyses, net photosynthetic rate and chlorophyll a fluorescence (JIP test) were measured 21 days after transplanting the plants to ex vitro conditions. The obtained results showed that the plants, acclimatized ex vitro in the substrate with Lumbrical, presented better growth (stem length, number of leaves, leaf area and fresh mass) and photosynthetic characteristics as compared to the control plants. This biostimulator could also be used to improve acclimatization in other woody species

Stomatal Morphology and Desiccation Response of Persian Walnut Tissue Culture Plantlets Influenced by the Gelling Agent of In Vitro Culture Medium

There are diverse types of gelling agents that are used in media cultures. Agar and Gelrite are among the gelling agents used in DKW culture medium, as the common culture medium for the micropropagation of walnut plants. The effects of these gelling agents have not been investigated on the successful production of in vitro explants and the response of the explants to ex vitro evaporative demand is overlooked so far. Stomata and water relations of tissue culture medium determine the successful production of in vitro plants, therefore this experiment was conducted to investigate the effect of two types of gelling agents (Agar and Gelrite) on the stomatal characteristics, transpiration rate (E), and desiccation responses of in vitro walnut explants. Stomatal morphology, transpiration rate, RWC, and some morpho-physiological traits such as shoot length, chlorophyll content, osmotic potential (ψs), proline, and glycine betaine content were evaluated in micropropagated walnut explants cultured on Agar or Gelrite. Analysis of results indicated no considerable changes in the morpho-physiological characteristics of explants grown in DKW medium containing Agar or Gelrite gelling agents. Compared with the medium containing Agar, adding Gelrite to the DKW medium caused a decrease in E and an increase in relative leaf water content (RWC) of the walnut explant's leaves during desiccation. Gelrite induced generation of more closed stomata leading to a reduction in E and increase in RWC during desiccation. This resulted in improvement of walnut plantlet's capacity to conserve their water content and as the consequence promoted ability to prevent ex vitro wilting.

Silver nanoparticles as the sterilant in large-scale micropropagation of chrysanthemum

Micropropagation has proven to be an effective method for large-scale plant production in a short time and a useful tool for plant breeding. Microbial contamination is one of the most difficult micropropagation challenges, resulting in reduced plant quality and loss of valuable stocks. Therefore, sterilization of culture media is a critical step in plant micropropagation. However, sterilized media might reduce the activity of plant growth regulators and nutritional components of culture media. The sterilization effects of silver nanoparticles (AgNP) on the growth of explants and culture media were examined. The treatment with 250 ppm AgNP for 15 to 20 min of 4-wk-old ex vitro leaves proved optimal for controlling the contamination. Furthermore, the Murashige and Skoog medium containing 4 ppm AgNP resulted in 100% medium disinfection (no contamination) after 4 wk of culture. The plantlets obtained from non-sterilized MS medium (NoM) containing 4 ppm AgNP and 4 g L−1 agar gave similar results as the control medium with 8 g L−1 agar and the absence of AgNP. Large scale culture systems using NoM in large plastic containers of two different sizes (NoM1 and NoM2) could produce quality plantlets. Chrysanthemum plantlets in the NoM1 system showed higher antioxidant enzyme activities of ascorbate peroxidase and superoxide dismutase than plantlets in the autoclaved medium. Furthermore, the plantlets from NoM were better acclimatized under greenhouse conditions than those from the autoclaved medium (AuM) system. The developmental stages (flower buds and blooming time) of NoM1 and NoM2 plantlets, were 1 wk earlier than those from the AuM system. The successful use of AgNP as a sterilizer and as a component of culture media would reduce the cost of micropropagation and improve plants' quality.

Micropropagation of banana (Musa spp) using temporary immersion bioreactor system

Banana is an important crop in the tropics which possess the potential for commercial production in Nigeria. Large scale production requires large volume of planting materials which may be difficult to obtain using conventional methods of propagation. Temporary immersion bioreactor system (TIBs) is a cost effective method for micropropagation of plants. The present study was carried out to develop an efficient method for rapid multiplication of banana using temporary immersion bioreactor system (TIBs). Banana microshoots were regenerated from young suckers obtained from field grown plants using conventional plant tissue culture. Microshoots of 2cm length were used as explants for multiplication in temporary immersion bioreactor system. Ten (10) explants were cultured in bioreactor bottles containing Murashinge and Skoog (MS) liquid media supplemented with different concentrations of 6-bezylaminopurine (BAP) with or without 250mg/L Activated Charcoal (AC). Results showed that explants cultured in media supplemented with 2 mg/L or 1mg/L BAP without AC gave the highest shoot multiplication rate of 900% and 800%, respectively compared to hormone free media. Production of competent plants (plants ready for ex vitro establisment) were however, influenced by the presence of AC and the highest percentage of competent plants (80%) were produced when media was fortified with 1mg/L BAP+ 250mg AC. Regenerated plants were successfully established in the field and were morphologically normal and fertile.

Micropropagation of the therapeutic-honey plants

Demand for therapeutic honey is driving establishment of Leptospermum plantations. This study developed micropropagation methods for two species – Leptospermum polygalifolium Salisb. and L. scoparium J.R.Forst. & G.Forst. The study determined how shoot proliferation and adventitious rooting were influenced by the original explant position on the seedling and the concentration of benzyladenine (BA) in the proliferation medium. Hormone-free node culture was highly effective for both species. Nodal explants often formed roots in the absence of BA and developed elongated axillary shoots. Median shoot numbers of 584 and 659 were formed in 31–32 weeks from a single L. polygalifolium or L. scoparium seed, respectively. A low BA dose was effective for callogenesis and shoot proliferation of L. polygalifolium, but not L. scoparium. The median number of shoots produced from a single L. polygalifolium seed was 630 using 2.22-μM BA. This dose induced extremely high shoot numbers in some clones because explants often produced extensive callus and multiple short shoots. Shoots formed adventitious roots without indole-3-butyric acid and plantlets were acclimatised to nursery conditions. The original explant position did not influence shoot proliferation or adventitious rooting. Leptospermum polygalifolium and L. scoparium proved amenable to micropropagation, facilitating rapid establishment of nectar plantations.

Optimization of stages of clonal micropropagation of garden strawberry varieties Nelly and Kemiya breeding of NCFSCHVW

The production of strawberry planting material using the method of clonal micropropagation of plants is promising, modern and already widely used. This paper presents the results of assessing the breeding potential of garden strawberry varieties Nelly and Kemiya (selection of FSBSI NCFSCHVW) in in vitro culture. Apexes from growing rosettes of strawberries served as the starting material for the initiation. The best survival rate of explants was noted during the initiation in the second decade of August, which were in Nelly variety - 90%, Kemiya - 83.3%. At the stage of proliferation, 6-Benzylaminopurine (6-BAP) was injected into the culture medium at various concentrations (0; 0.5; 1.0 and 1.5 mg / L). To increase the efficiency of micropropagation of strawberries Nelly and Kemiya varieties, it is advisable to add 6-BAP into the culture medium with the amount of 1.0 mg / l. In this variant, 10.6 shoots were formed in the explants in Nelly variety, 5.9 in Kemiya variety. With an increase of the concentration of 6-BAP to 1.5 mg / l, the number of vitrified shoots were raised. According to the results of three subcultures, it was noted that the Nelly variety has a higher reproduction potential compared to the Kemiya variety. At the stage of rhizogenesis, the addition of auxins to the medium was not required, since the roots of regenerated plants formed independently.

The effectiveness of the use of the silicon-containing preparation Siliplant in the case of clonal micropropagation of the rose Reine Sammut

Clonal micropropagation of plants is a modern popular method of vegetative reproduction, which allows to obtain high-quality planting material in a large volume in a short time. The research is devoted to the technology of clonal micropropagation of the rose Reine Sammut, in particular, optimization of the nutrient medium composition. To increase the efficiency of micropropagation, the silicon-containing preparation Siliplant was added to the composition of the nutrient medium according to the Murashige-Skoog (MS) recipe as an alternative to the complex of trace elements. At the stage of the actual micropropagation with the combined use of 6-benzylaminopurine (6-BAP) at doses of 1.0; 2.0 and 3.0 mg/l and Siliplant at doses of 1.0; 2.0 and 3.0 ml/l, no large differences were found. Nevertheless, the inclusion of Siliplant (1.0 ml/l) and 6-BAP (3.0 mg/l) in the medium contributed to an increase in the reproduction factor by 3.1 pcs./cuttings at LSD05= 1.0 pcs. In addition, Siliplant at a dose of 1.0 ml/l as part of a hormone-free nutrient medium MS contributed to a significant increase in shoot growth (by 4.2 mm at LSD05= 3.6), and at a dose of 3.0 ml/l stimulated the development of standard micro-plants of roses.

A review on in-vitro micropropagation of agave and other plants

This article gives a brief review on research progress on the use of tissue culture in the micropropagation of agaves and a few economic plants. Fragments of leaf meristem, rhizomes, meristematic apex, root and excised embryo are used as explants in MS medium. Techniques used in the micropropagation of several species of Agave gave promising results. In vitro propagation is extensively used for propagation of Agave spp. and native plant species in some established centers in some countries. In vitro somatic embryogenesis is considered as an important pre-requisite for genetic improvement, as well as for mass propagation. The asceptic mass production of callus is efficiently used in the extraction of secondary metabolites of medicinal use. The technique is recommended specially for the propagation of perennial native economic plant species like Agave which takes more than eight years to reach reproductive stages and produces seeds. The massive production of micropropagated native plants could be effectively used in the reforestation of the species in their natural habitats.

The effect of different compositions of growth media on the development of microplants of the Antonina potato variety

The results of studying the influence of growth media of different compositions on the growth and development of healthy microplants of potato variety Antonina in laboratory conditions in vitro are presented. Six compositions of Murashige and Skoog culture medium were considered: standard according to the recipe modified for micropropagation (control); with a reduced content of mineral components (up to 1/2 and 1/3); with an increased content of agar-agar (10 g/l); with a reduced content of agar-agar (4 g/l), and modified by the addition of 3 mg/l giberrellic acid and 1 mg/l indoleacetic acid. The following parameters of cultivated plants were studied: plant length, root presence, number of internodes, total plant weight, leaf weight, root weight, leaf surface area. The use of growth media with a reduced content of mineral components led to an increase in the length of the plants under study by 8–9%, a decrease in the number of internodes by 0.49–0.85 pcs/plant, an increase in the mass of the root system by 23% and a decrease in the mass of the shoot due to reduced leaf mass by 16–31%, as well as a decrease in the total leaf surface area by 21–30%. When growing plants on a growth medium with an increased content of agar-agar, the following changes were observed: a decrease in the length of plants by 13%, a decrease in the mass of the root system by 8%, a decrease in the shoot mass due to reduced leaf mass by 11% and stem mass by 17%. Plants grown on a growth medium with a low content of agar-agar had a shorter stem length (by 12%), a lower number of internodes (by 0.93 pcs/ plant), a lower mass of root system and leaves by 31% and 11%, respectively, compared to the control. In this variant, the rate of rhizogenesis was also reduced. With the addition of giberrellic and indoleacetic acid to the growth medium, the plants demonstrated a significant increase in length by 32%, a decrease in the root mass by 77%, and a decrease in the shoot mass due to reduced leaf mass by 58% and stem mass by 33%. The total leaf surface area was 34% lower than the control values. In order to accelerate micropropagation of healthy Antonina potato plants and prepare the plants for replanting to an aero-hydroponic system, Murashige and Skoog medium modified for micropropagation proved to be the best option among the tested growth media. In order to keep this variety in the in-vitro collection, it is recommended to use the MS growth medium with a low agar-agar content.

NANOPARTÍCULAS DE PLATA EN EL ESTABLECIMIENTO in vitro DE ÁPICES DE GLADIOLO

La micropropagación de plantas puede ser limitada por varios factores, como la contaminación in vitro. La desinfección en la etapa de establecimiento es uno de los pasos más importantes en la prevención de la contaminación bacteriana y fúngica. Las nanopartículas de plata (NPsAg) son un material que muestra alta capacidad en la eliminación de microorganismos. El objetivo de esta investigación fue evaluar el efecto de las NPsAg como auxiliar en la desinfección de ápices de gladiolo y en la desinfección del medio de cultivo. Las NPsAg se aplicaron como auxiliar en la desinfección de explantes, en cuatro concentraciones (25, 50, 100 y 200 mg L-1) y se combinaron con cuatro tiempos de inmersión (5, 10, 15 y 20 min). Por otro lado, las NPsAg se utilizaron en cinco concentraciones (0, 25, 50, 100 y 200 mg L-1), como parte del medio de cultivo para evitar el crecimiento de microorganismos. Con 50 mg L-1 de NPsAg se obtuvo el mayor porcentaje de asepsia (91.67 %) y la mejor longitud del brote (28.62 mm). El tiempo de inmersión de 5 a 20 min de los explantes en NPsAg no tuvo efecto en la asepsia de los mismos, pues el resultado fue similar en los cuatro tiempos de inmersión. El análisis de interacción de factores sugiere que 50 mg L-1 durante 15 min fue el mejor tratamiento. La desinfección del medio de cultivo con 25, 50, 100 y 200 mg L-1 de NPsAg generó 100 % de asepsia, en tanto que el control, sin NPsAg, mostró 80 % de contaminación con microorganismos. El uso de 50 mg L-1 generó una longitud del brote 35.5 % mayor, en comparación con el control sin NPsAg.

INFLUENCE OF THE STERILIZATION METHOD AND THE TIME OF INTRODUCTION INTO IN VITRO CULTURE ON THE VIABILITY OF EXPLANTS OF THE CLONAL APPLE STOCK 54-118

The stage of introducing explants into a sterile culture is difficult in the technology of clonal micropropagation of plants. The article shows the possible ways of sterilization and the introduction terms of explants of the clonal apple stocks 54-118 into in vitro culture in order to reduce planting infection and increase the yield of sterile viable explants. The best time for introducing clonal apple stocks 54-118 into sterile in vitro culture is the period of active shoot growth. Sterilization of explants with ethyl alcohol (70,0 %, 1 min) in combination with hydrogen peroxide (33,0 %) for 7 minutes and ethyl alcohol (70,0 %, 1 min) in combination with diacide (0,1 %) within 6 minutes contributed to the production of 63,0 % and 60,0 % of viable sterile explants.

True-to-type micropropagated plants of para rubber (Hevea brasiliensis Müll. Arg.) via somatic embryogenesis

Plant micropropagation via somatic embryogenesis is a powerful technique for rapid mass propagation, especially in para rubber (Hevea brasiliensis Müll. Arg.). However, somaclonal variations are the major limitation of this process. To date, DNA fingerprinting, i.e., RAPD (Randomly Amplified Polymorphic DNA), Star Codon Targeted (SCoT), and SSRs (Simple Sequence Repeats), is one of the most successful technologies to detect the genetic fidelity in the somatic embryos. The aim of present study was to induce somatic embryos from inner integument explants of para rubber cv. ‘RRIM 600’ at different developmental stages and subsequent acclimatization and transplantation (under greenhouse and field conditions) of the propagated seedlings. The genetic stability of the plants derived from somatic embryos was also analysed in comparison to the mother plant using RAPD, SCoT and SSRs markers. Somatic embryos derived from inner integuments of 5-week-old immature seeds after pollination were more efficient than older and younger seeds. In addition, para rubber mother plants cv. ‘RRIM600’ and plants derived from somatic embryogenesis demonstrated the same pattern of DNA fragments, as confirmed by three PCR-based techniques, RAPD, SCoT and SSRs, whereas these in the pattern were different from ‘RRIT 226’, ‘PB 235’, ‘PB 251’, ‘PB 255’ and ‘BMP 24’. Interestingly, T2 plant was found to possess somaclonal variations when compared with mother plant. Based on the results, we confirm that the plants derived from somatic embryogenesis of para rubber cv. ‘RRIM 600’ were true-to-type to that of ‘RRIM 600’ master stock.

The role of phytohormones in the induction of morphogenesis by Calycanthus floridus L. explants for in vitro propagation

Aims. Induction of morphogenesis in vitro by the cultivation of explants on nutrient media depends on many interrelated factors. The main inducers and coordinators of morphogenic processes in vitro are phytohormones, which are introduced into nutrient media in certain concentrations and play a major role in regulating the growth and development of explants. Therefore, elucidation of the morphogenic potential dependence of explants of Clycanthus floridus L. the effect of exogenous growth regulators (phytohormones) is determined by the aim of the research. Materials and methods. The work was conducted in the Plant Micropropagation Laboratory of the National Dendrological Park "Sofiyivka" of the National Academy of Sciences of Ukraine. Plant material C. floridus was used for research obtained from seeds sprouted in vitro. To determine the efficiency of growth, development, and morphogenic potential of C. floridus explants used basal nutrient media according to Murashige and Skoog (MS) and Driver and Kuniyuki Walnut (DKW). Morphogenesis was stimulated by exogenous phytohormones: 6-benzylaminopurine (BAP) – 0.1–2.0 mg/l, β-indolylbutyric acid (IBA) – 0.01–1.00 mg/L and cysteine – 0.5–2.0 mg/l. Results and discussion.The data obtained during the experiment showed that the growth, development and increase of the morphogenic potential of explants, and, accordingly, the multiplication coefficient, largely depend on the content and concentrations of phytohormones in nutrient media. In explants C. floridus cultured on nutrient media according to Murashige and Skoog, modified with the addition of 0.5 mg/L 6-BAP and 0.08 β-IBA and Driver and Kuniyuki with a content of 0.5 mg/L 6-BAP; 0.08 mg/l β-IBA and 1.0 g/L cysteine, active morphogenesis processes and an increase in the multiplication coefficient were detected. Conclusions. The morphogenic capacity of C. floridus explants was investigated. It was found that the active passage of morphogenesis processes in explants and an increase in the multiplication coefficient indicators were facilitated by modification of nutrient media prescribed by Murashige and Skoog with the addition of 0.5 mg/L 6-BAP and 0.08 β-IBA and Driver and Kuniyuki with the content of 0.5 mg/L 6-BAP; 0.08 mg/l β-IBA; 1.0 g/L cysteine.

Farm or lab? Chamazulene content of Artemisia arborescens (Vill.) L. essential oil and callus volatile metabolites isolate

Plant micropropagation has been proposed as a promising solution for the improvement of medicinal plants and the commercial production of phytochemicals. Tree Wormwood (Artemisia arborescens, Asteraceae) has been identified as a promising source for chamazulene production, a phytochemical compound with anti-inflammatory activity. However, the concurrent presence of β-thujone, an acute toxic agent is a significant limitation of Tree Wormwood utilization in essential oil (EO) production. Present study is aiming to develop in vitro cultivation protocols of tree wormwood and study the effects of plant growth regulators (PGRs) and culture conditions on the callus production of volatile metabolites (VM) and its equilibrium of chamazulene and β-thujone. Three treatments all containing the auxin 1-Naphthaleneacetic acid (NAA), and each one respectively the cytokines Kinetin (K), 16-Benzylaminopurine (BN), and Thidiazuron (TD) in Murashige and Skoog (MS) basal media, were applied under constant dark (D), while K and BN were also grown under light (L). Both EO and VMs, retrieved from the naturally and laboratory grown biomass were analyzed by a Gas Chromatographer-Mass Spectrometer (GC–MS) coupled with Gas Chromatographer-Flame Ionization Detector (GC-FID). The biotechnologically produced EOs presented a significantly increased yield per area and time, while callogenesis PGRs combinations and culture conditions exhibited promising results on the induction of chamazulene biosynthesis.