Micropropagation Method Research Articles

A comparative microbiological study of the essential oils from the genus Rosa L.

The medicinal plant raw material containing essential oils always attract the attention of scientists from various countries worldwide. The search for new sources of the plant raw material rich in biologically active substances, particularly volatile compounds, remains a relevant task in modern pharmaceutical science. Our focus has been drawn to Rosa damascena Mill. of the “Veselka” (“Rainbow”) variety grown in the natural environment of Zaporizhzhia; its essential oil exhibits anti-inflammatory, antimicrobial, and antiseptic effects. To obtain a valuable medicinal plant raw material from Rosa damascena with a high content of essential oil, it was introduced into in vitro culture to produce aseptic regenerants, followed by their reintroduction into the natural environment. Aim. To conduct a comparative microbiological study of essential oils from Rosa damascena of the “Veselka” variety grown via clonal micropropagation in in vitro culture and in the natural environment of Zaporizhzhia. Materials and methods. The study objects were essential oils obtained by water distillation from rose petals cultivated using the method of clonal micropropagation in in vitro culture (at the premises of the Educational and Scientific Medical Laboratory Center with a vivarium) and the natural environment of Zaporizhzhia. The study of the antimicrobial activity was conducted in the microbiological laboratory of the Department of Microbiology, Virology, and Immunology at Zaporizhzhia State Medical and Pharmaceutical University. Results and discussion. The essential oils from Rosa damascena grown via clonal micropropagation in in vitro culture and under natural conditions in Zaporizhzhia effectively inhibited the growth of E. coli (21.3 mm and 12.0 mm, respectively) and S. aureus (11.3 mm and 10.2 mm, respectively). The results of the study on C. albicans indicate a high antifungal activity in both essential oils; the mean inhibition zone diameter in the experiments using the oil from Rosa damascena cultivated in in vitro culture is less: 33.3 mm compared to 40 mm in the experiments with the oil from roses grown in the natural environment of Zaporizhzhia. Conclusions. The method of clonal micropropagation of Rosa damascena of the “Veselka” variety is effective for obtaining a large amount of planting material in a short time with the subsequent reintroduction into the natural environment to expand the raw material base of valuable medicinal plants with a high content of essential oils. The results of the study of the antimicrobial activity of the essential oil of Rosa damascena of the “Veselka” variety cultivated through clonal micropropagation in in vitro culture have exhibited a high antimicrobial activity and moderate antifungal activity compared to the essential oil from Rosa damascena grown in the natural environment of Zaporizhzhia. Therefore, Rosa damascena Mill. of the “Veselka” variety cultivated under in vitro conditions is a promising source of essential oil with a high antibacterial effect in order to create new herbal formulations.

Ecophysiological Recovery of Micropropagated Olive Cultivars: Field Research in an Irrigated Super-High-Density Orchard

This research focuses on the seasonal patterns of the ecophysiological recovery of four olive cultivars (Arbequina, Coratina, Frantoio, and Urano), both micropropagated and self-rooted, grown in a mature, irrigated, super high-density (SHD) orchard under Mediterranean conditions (Southern of Italy). The aim was to observe the impact of the micropropagation method on the ecophysiological responses. Ecophysiological parameters, including leaf water potential (LWP), stomatal conductance (gs), net photosynthetic rate (Pn), and transpiration rate (E) were assessed. Self-rooted trees consistently exhibited superior gs, E, and Pn recovery compared to the micropropagated ones. ‘Arbequina’ maintained elevated levels of Pn under water-deficit conditions. ‘Coratina’ exhibited increases in gs and E after irrigation. ‘Frantoio’ demonstrated recovery capabilities, with lower LWP and higher Pn under stress. ‘Urano’ micropropagated trees achieved higher gs values in mid-summer, while self-rooted trees sustained higher Pn later in the season. This field research highlighted the important role of the propagation method in optimizing the physiological performance of olive cultivars in SHD orchards. Furthermore, it highlighted the necessity of long-term studies on the effects of propagation methods and their interactions with other farming practices.

Micropropagation of Vaccinium corymbosum L.‘Bluecrop’ in Rocker Temporary Immersion System (TIS) Bioreactor

Blueberries are high-value fruits. The traditional method of propagation by cuttings cannot supply the modern market with large quantities of seedlings. The method of micropropagation of plants in vitro makes it possible to bring the production of blueberry seedlings to the highest level. Blueberries have not been sufficiently studied in in vitro culture, so the search for the simplest and most cost-effective methods of micropropagation remains relevant. The problem of accelerated micropropagation of blueberries can be solved using rocker-type bioreactors, which differ from other models in terms of simplicity of design and low cost. A study was carried out to evaluate the effectiveness of micropropagation of Vaccinium corymbosum 'Bluecrop' in rocker bioreactors. Two types of bioreactors were compared: the bioreactor of the Platform system and the TIS rocker bioreactor modified by the author. As a control, blueberries were grown on a semi-solid medium. The effectiveness of blueberry micropropagation was evaluated by the following indicators: multiplication coefficient, shoot length, and proportion of vitrified shoots. Experiments were conducted on WPM medium, with zeatin supplementation at a concentration of 1.0 mg/l, resulting in optimal results. It is shown that the rocker bioreactor is slightly inferior to the plantform bioreactor in micropropagation but outperforms the method of micropropagation on semisolid media. The rocker bioreactor can be fully utilized for production purposes. In order to reduce costs and increase technical reliability, the working principle of the mechanical drive of the author's model of a rocker-type bioreactor was changed.

Features of clonal micropropagation of plum PK SK 1 clonal rootstock

Methodological features of the main clonal micropropagation stages of plum PK SK 1 clonal rootstock (of NCFSCHVW selection) are described. The rootstock is characterized by a high adaptability to soil and climatic conditions of the region. In particular, it exhibits good winter hardiness, resistance to overwatered soils, tolerance to drought, and the capacity to grow on heavy soils. Plum seedlings on this rootstock develop a powerful root system. These advantages render the development and improvement of methods for clonal micropropagation of plum PK SK 1 clonal rootstock highly relevant. The research was carried out in 2019-2022. The period of active shoot growth, i.e., the first and second decade of may, was established to be the most optimal period for introduction of explants into in vitro culture. In this period, regeneration reaches 78.1–93.8 %. Eff ective shoot multiplication proceeds on MS medium with Fe-EDTA or MS medium with Fe-EDDHA and 6-BAP in the amount of 1 mg/L, with the highest number of shoots formed from one explant following the fourth passage (on average, 11.7 and 12.3 shoots per explant). A signifi cant eff ect of the 6-BAP concentration on the number of shoots formed was observed. The combined eff ect of the form of chelated iron and 6-BAP concentration on the number of shoots formed was established only after 3-5 passages on MS medium with Fe-EDDHA. The rooting stage of PK SK 1 rootstock can also be carried out on a hormone-free MS medium with Fe-EDDHA. The rooting effi ciency reaches 57.1–61.8 %. On average, 2.4 roots are formed per shoot with a length from 1.2 to 5.1 cm, with the root quality being high (not cartilaginous and not brittle). For the adaptation stage, rooted shoots with a height of at least 2 cm having two or more roots with a length of at least 2-3 cm are selected. A mixture of the Agrobalt plant nutrient soil, vermiculite, and perlite in a ratio of 3:1:1 can be used as a substrate. Following four weeks of adaptation, the number of adapted plants averaged 87.5 %.

Optimization of clonal micropropagation of stone fruit crops

The aim of the research was to optimize the clonal micropropagation of stone crops through the use of improved techniques. The objects of the research at the stage of introduction into culture in vitro were meristematic apexes, at the stage of micro-propagation itself – micro-shoots, at the rooting stage – rooted micro-shoots, at the adaptation stage – micro-plants. All experiments were carried out according to generally accepted methods using the example of the steppe cherry variety Shchedraya, the domestic plum variety Kazanskaya and the hybrid sweet cherry Fatezh. As a result of the research, it was found that the use of a 10 % chlorhexidine solution for sterilization of the initial plant material of stone crops increased the survival rate of apexes on average by 2.4 %; the combined use of growth regulators 6-benzylaminopurine 0.5 mg/l, gibberellic acid 0.2 mg/l, indo-lil-3-butyric acid (IBA) 0.2 mg/l in a nutrient medium and an experimental diode pulsed phytoirradiator in the last proliferation passage activated the proliferation of micro-shoots, increasing the reproduction coefficient of stone crops in 6 passages by an average of 0.9 pcs/explant; the addition of 1.0 mg/l (IBA ) to the nutrient medium provided an increase in the rooting capacity of micro-shoots of stone crops by an average of 9.6 %. The treatment of micro-plants of stone crops at the stage of adaptation by spraying with an 8 % solution of the extract of the products of the larvae of the large wax moth and the use of an experimental LED pulsed phytoirradiator with a mixed spectrum contributed to an increase in their survival rate by an average of 12.4 %. Compliance with the proposed improved methods of clonal micropropagation of stone crops using growth regulators and an experimental LED pulse phytoirradiator with a mixed spectrum made it possible to increase the yield of standard adapted micro-plants by 2 times. At the same time, the cost of one adapted micro-plant decreased by an average of 11.4 rubles, and the profitability of obtaining improved stone crop material increased by 33.7 % and amounted to 160.7 %.

Micropropagation, genetic fidelity and chromatographic analysis in Evolvulus alsinoides (L.): A potent multipurpose medicinal plant

Evolvulus alsinoides (L.) is an annually growing medicinal plant used in many popular systems as a source of traditional medicine. However, fresh plant material for medicinal uses and research purposes is critically unavailable year-round due to its short life cycle. A micropropagation method for E. alsinoides was developed, and its genetic diversity was described. Surface sterilized nodal explants were inoculated onto Murashige and Skoog (MS) medium supplemented with different plant growth regulators; the best response of multiple shoot induction was observed on MS media augmented with 5 μM kinetin (Kn) and 0.1 μM Indole-3-acetic acid (IAA), which gave a maximum number of 10.27 ± 0.0118 shoots. Further, the addition of 30 μM of adenine sulfate to the MS media enhanced shoot multiplication (18). The cloned shoots were accomplished to in vitro rooting on MS medium with 0.1 μM Indole-3-butyric acid (IBA) and 10 μM phloroglucinol, resulting in 16 healthy roots. The rooted plantlets were acclimatized in the shade house. The genetic stability of micropropagated plants was confirmed through molecular marker analysis. Gas chromatography-mass spectrometry (GC-MS) analysis of the tissues of clones revealed the contents of secondary metabolites. High-Performance Thin Layer Chromatography (HPTLC) revealed that squalene levels were highest in plants cultured in MS medium supplemented with adenine sulfate (378.89 ng/ 15 μl). The methods developed and defined are highly reproducible and can be used for commercial production of E. alsinoides valued for human health.

Ex-situ Conservation of Bulbophyllum leopardinum, A Threatened Medicinal Orchid of Nepal

A successful micropropagation method was developed via the in-vitro seed germination and seedling growth of the epiphytic and/or lithophytic orchid Bulbophyllum leopardinum, a species having horticultural and therapeutic significance. To enhance seed germination, several quantities and combinations of naphthalene acetic acid (NAA), 6-benzyl amino purine (BAP), indole acetic acid (IAA), gibberellic acid (GA3), and coconut water (CW) were added to 0.8% (w/v) agar-solidified MS medium. Half-strength MS medium has been experimented with alone and in combination with BAP, Kinetin (Kn), and GA3 to promote shoot development. In-vitro-developed healthy shoots were chosen to establish roots in a half-strength MS (HMS) medium supplemented with various auxins. The best and earliest seed germination with the greenest protocorms (96.3±0.5% in 7 weeks) was achieved on HMS medium fortified with 15% CW (H15C). Further tests for the shoot as well as root development were continued with an H15C medium. H15C with 1 mg/l BAP and 1.5 mg/l Kinetin was most effective for early in vitro development and differentiation into seedlings with the many long shoots (9.3±0.1 shoots and 2.4±0.1 cm per culture) within 12 weeks of sub-culture. The most suitable rooting hormone was 1 mg/l NAA (4.2±0.26 roots per culture). This medium also produced the longest roots (1.9±0.09 cm per culture). By successfully developing a protocol for the mass propagation of B. leopardinum, this research has enhanced both the cultivation and the commercialization potential of this species.

Effect of Different Plant Growth Regulators on In-vitro Micropropagation of Banana Cultivar Grand Naine (Musa spp.)

Banana (Musa sapientum) is one of the most important fruit crop cultivated in India. It is widely grown in more than 150 countries, producing on an average of 105 million tons of fruit per annum. Grand naine Banana variety is the most commonly cultivated banana with high yielding plantain. This variety mainly grown in the region of Uttar Pradesh state. Present study mainly focuses on the in-vitro micropropagation method which is described for banana cv. Grand naine. For this study, healthy sword suckers of three months old banana plantlets was used as explants. The explants was sterilized thoroughly and cultured on MS (Murashige and Skoog) media supplemented with the different concentration and combination of auxin and cytokinin like BAP, IAA and NAA which were used for the optimization of best plant growth regulator combination for multiple shoot generation and shoot growth. Among all the tested cytokinin 5 mg/l BAP along with 1mg/l IAA in medium concentration performed well and produced healthy shoots further at higher concentration of all tested cytokinin showed declining effect within the growth parameters. For the rooting purpose, MS media was supplemented with different levels of IBA, IAA and NAA. Results indicated that rooting was highly promoted in the medium supplemented with 0.5 mg/L IAA + 1.5 mg/L IBA.

Prospects for the creation of new phytochemical medicinal products based on Myrtus communis L. (a review)

The number of medicinal products based on biologically active substances (BAS) of plant origin is increasing every year. Scientists are exploring non-official plant raw materials used in non-traditional medicine to expand the raw material base for creating new phytopreparations. Among numerous plants, common myrtle (Myrtus communis L.), which is a fairly common indoor plant, is recognized as a valuable source of BAS. The aim of the work is to analyze and summarize new literature data on the botanical characteristics, chemical composition, pharmacological and toxicological properties of Myrtus communis L. Materials and methods. Data were collected from several legitimate databases and services such as PubMed, Medline, Scopus, Google Scholar, ResearchGate using the key terms “Myrtus communis L.”, “botanical characteristics”, “pharmacological studies”, “phytochemical studies” in each database data Relevant and related data were filtered as appropriate if deemed relevant to the topic of interest. Time frames have also been adjusted to provide up-to-date information. Results. Myrtus communis L., commonly known as common myrtle, is a widely recognized medicinal plant in the Mediterranean region. Various parts of this plant, including the leaves, fruits, and flowers, have been used in traditional medicine for treating a wide range of ailments. The pharmacological properties of common myrtle are attributed to the presence of essential oils and phenolic compounds, and these properties include antimicrobial, antiviral, antidiabetic, antispasmodic, vasodilator, antiulcer, antioxidant, antitumor, and anti-inflammatory effects. This article offers an overview of the botanical characteristics and chemical composition of different parts of common myrtle, and it presents a literature review of pharmacological studies that provide evidence for the plant’s medicinal properties. Conclusions. Common myrtle is a promising plant for further pharmacognostic research with the aim of expanding the raw material base, using the method of clonal micropropagation and creating new phytopreparations based on it.

First Ex Situ In Vitro Propagation Protocol of Coronilla viminalis Salisb., An Endangered Fodder Species Adapted to Drought and Salinity

Coronilla viminalis Salisb. is a fodder leguminous plant from the Canary Islands and Northwestern Africa with adaptation to drought. Its conservation status is critical. Its low germination capacity and intense loss of viability of the seeds over time complicate its conservation, limiting its long-term maintenance in germplasm banks and hampering its potential use as a resilient fodder crop. Therefore, in this work, two alternative propagation methods have been addressed. The first was based on facilitating seed germination under aseptic conditions, supplementing the media with gibberellic acid (GA3). The latter consisted of creating a micropropagation method from nodal segments and testing different media and pretreatments with indole-3-acetic acid (IAA). The quantity and quality of the roots were assessed, and the plant acclimation rate was measured. The results showed that the optimum concentration of GA3 for germination, up to 98%, was 250 PPM. The experiments showed that using IAA in the micropropagation media is critical for in vitro rooting in this species. A hormonal pretreatment with IAA significantly improved the rooting efficiency compared to supplementing it into the culture media. One hundred percent of acclimated plants survived the process. These new protocols will help conserve the species and explore its possibilities as fodder crops.

Efficiency-cost, micropropagation, bioreactors, temporary immersion systems

Objective: to estimate the production price of strawberry “mother” plant of the CP-Jacona variety in both TIS and TS, and to compare these prices with the price of the imported “mother” plant. Design/Methodology/Approach: micropropagation methods have been used as an effective means for the mass production of pathogen-free plants, in small spaces and relatively short periods of time. In particular, in vitro Temporary Immersion Systems (TIS) applied to the production of strawberry “mother” plants have been shown to offer technological and quantitative benefits, as well as a higher proliferation rate, compared to the in vitro Traditional System (TS). Despite the benefits of TIS, these systems have not been evaluated in terms of the price at which the “mother” plant can be produced and whether it is a profitable option to supply strawberry producers. The traditional method of financial analysis was applied. Results: the price of the CP-Jacona variety plant obtained from the third production period through TIS was lower than the price of the imported Festival or Camarosa varieties. Findings/Conclusions: both the high production rate and the low rate of loss from handling in TIS were fundamental aspects to obtain lower prices than those of imported varieties.

Efficient In Vitro Platform for Multiplication, Acclimatization, and Deliver of High-NaCl-Tolerant Clones of the Halophyte Arthrocaulon macrostachyum

Halophytes are species able to grow and complete their life cycle under NaCl concentrations above 0.2 M. In a context of growing population and uncertain agricultural food sustainability, there is an interest on halophytes as potential source of food and fodder. However, scarce research has addressed in vitro propagation of halophytes as a tool for study, conservation, and propagation of elite germplasm. In this study, an efficient method for micropropagation of the succulent and salt accumulator halophyte Arthrocaulon macrostachyum has been established for the first time, using shoot tips from in vitro-germinated plant material. During shoot multiplication stage, superior genotypes were selected from explants grown in high strength and NaCl content medium and subsequently rooted and acclimatized to ex vitro conditions. A comprehensive characterization including determination of oxidative stress, photosynthesis performance, and mineral nutrient contents was done. This research gains insight into the physiological and biochemical characterization of halophytes during micropropagation and provides a solid platform for the production of elite A. macrostachyum germplasm for ulterior uses.

Assessment of genetic stability of micropropagated Curculigo latifolia from Indonesia by RAPD and ISSR

Micropropagation of Curculigo latifolia is very important because this plant has high economic value including containing Curculin and Neoculin as sweetener and taste-modifying proteins. Micropropagated plants may experience genetic changes like somaclonal variation so it is necessary to assess their genetic stability. The purpose of this study was to obtain a micropropagation method of C. latifolia that is more efficient than the previous research and to assess the genetic stability of micropropagated C. latifolia. The plant material used was sterile leaves obtained from sterile seedlings raised from C. latifolia seed germination. Leaves were grown on a callus induction medium containing 5 mg l−1 indole-3-butyric acid and 3 mg l−1 BA in a dark room. Callus formed were subcultured to growth selection medium and shoots formed were subcultured to Murashige and Skoog-free growth regulator medium with the addition of 6 g l−1 agar and 3% sucrose. The cultures were incubated at 23ºC ± 2ºC under 1,500 lux white LED (PhilipTM TL) on a 16 hour/8 hour light/dark photoperiod. The cultures were incubated for 14 weeks. Genetic stability of in vitro propagated C. latifolia from Indonesia was assessed by molecular markers, RAPD and ISSR. The micropropagation method produces an efficient protocol because each growth stage occurs in a free growth regulator medium except for callus induction. Based on the results of the assessment using molecular markers, no polymorphism in DNA amplification using RAPD and a high percentage of genetic stability (66.67%) from ISSR analysis were obtained. In sum, the in vitro micropropagation protocol created can be used commercially to produce C. latifolia on a large scale that is genetically stable.

ВЛИЯНИЕ ТЕХНОЛОГИИ ПРОИЗВОДСТВА МАТОЧНЫХ РАСТЕНИЙ ВИНОГРАДА НА ИХ СПОСОБНОСТЬ К ВЕГЕТАТИВНОМУ РАЗМНОЖЕНИЮ

For many years, grapes were considered to be an unpromising crop in the conditions of the Central Non-Black Earth Zone but their wider spread has been encouraged by the appear- ance of new varieties whose berries ripen in a relatively short time. The assortment of modern grape varieties for amateur cultivation in the non-Black Earth Zone consists mainly of interspecific hybrids based on Vitis amurensis Rupr., Vitis riparia Michx., Vitis labrusca L., Vitis berlandieri Planch., which causes problems with their vegetative propagation. The article is devoted to the study of the effect of the method of obtaining planting material from vine mother plants on the increase of the regenerative capacity of the singletree cuttings obtained from them. During the threeyear research on the grape varieties Kishmish No. 342 and Moskovskiy Beliy, experiments were carried out on the additional growth of mother plants propagated by green and singletree cuttings and by the method of clonal micropropagation. As a result of the research, the advantage of using the technology of clonal micropropagation in the production of seedlings for the establishment of vine mother plantations was revealed. At the same time, the variety Kishmish No. 342 showed reliable differences from the control in terms of development indicators from the 2nd year of cultivation, and the variety Moskovskiy Beliy – from the 3rd year. Both varieties received 1.8–2.1 times more standard seedlings from mother plants propagated in vitro for three years on the basis of high rooting ability of singletree cuttings and the proportion of seedlings according to GOST 31783–2012.

Using Mathematical Optimization Models to Improve the Efficiency of Duckweeds (Wolffia arrhiza and Lemna minor) Micropropagation.

The method of plant micropropagation is widely used to obtain genetically homogeneous and infection-free plants for the needs of various industries and agriculture. Optimization of plant growth and development conditions plays a key role in economically successful micropropagation. Computer technologies have provided researchers with new approaches for modeling and a better understanding of the role of the factors involved in plant growth in vitro. To develop new models for optimizing growth conditions, we used plants with a high speed of vegetative in vitro reproduction, such as duckweed (Wolffia arrhiza and Lemna minor). Using the development of the optimal modeling of the biological processes, we have obtained the prescriptions for an individually balanced culture medium that enabled us to obtain 1.5-2.0 times more duckweed biomass with a 1.5 times higher protein concentration in the dry mass. Thus, we have demonstrated that the method of optimization modeling of the biological processes based on solving multinomial tasks from the series of quadratic equations can be used for the optimization of trophic needs of plants, specifically for micropropagation of duckweeds in vitro.

ПЕРСПЕКТИВЫ ИСПОЛЬЗОВАНИЯ РОДОДЕНДРОНОВ В ГОРОДСКОМ ОЗЕЛЕНЕНИИ И ВОЗМОЖНОСТИ ПРИМЕНЕНИЯ БИОТЕХНОЛОГИЧЕСКИХ МЕТОДОВ ДЛЯ ИХ МАССОВОГО РАЗМНОЖЕНИЯ

The article assesses the use of introduced representatives of the genus Rhododendron L. in urban landscaping in central Russia. The species and varietal diversity of rhododendrons allows us to meet the highest demands of specialists in green construction and landscape architecture. Rhododendrons are a difficult-to-root crop, so there is a constant search for means and methods that can increase the yield of planting material. Along with traditional methods of plant propagation, the method of clonal micropropagation is successfully used. The prospects of biotechnological methods for mass propagation of rhododendrons are shown.

Use of rocker bioreactor for propagation of the Vaccinium corymbosum L.

Plant biotechnology has the potential to meet the market demand for a significant number of seedlings. The current state of the industry is characterised by exemplary results in the field of micropropagation, with notable achievements observed in the cultivation of diverse plant crops, particularly berries. Nevertheless, certain aspects of this technology remain imperfect at the industrial level. The multiplication of microshoots of berry crops remains a costly process. The transition of numerous laboratories from semi-solid agar medium to liquid has resulted in a reduction in the costs associated with propagation. However, the majority of standard bioreactors are costly and frequently challenging to maintain. In the present study, a rocker-type bioreactor system for micropropagation of blueberries was tested using Vaccinium corymbosum L. ‘Pink Lemonade’ as an example. In comparison to the traditional method of micropropagation on a gel medium, the rocker-type bioreactor demonstrates superior outcomes. The multiplication coefficient of blueberry shoots was found to be approximately twofold higher when the liquid WPM medium was supplemented with 0.2 mg/l zeatin within the bioreactor, in comparison to the control variant cultivated on a semi-solid medium of identical composition. The shoots obtained in the bioreactor successfully completed the stages of rooting and acclimatisation to post-aseptic conditions.

Introduction and breeding of shadbush in Russia and abroad

Shadbush is one of the underutilized berry crops. Originally, it was an ornamental crop, but now it is increasingly used as a food crop. Numerous species of shadbush belong to the genus Amelanchier Medik.; wild species grow in North America, Europe, as well as in Western and Eastern Asia, and each region has different species composition. The year of 1590 marked the beginning of introduction of wild shadbush species with the most valuable features to other countries. In the 19th century, the introduction of shadbush became most popular. In 1800, shadbush breeding was launched in Canada, and then in the USA. In 1937, the production of commercial varieties was established. At present, in Russia, the most productive work on introduction and breeding of shadbush is carried out in Michurinsk (All-Russian Scientific Research Institute of Horticulture named after I.V. Michurin), Novosibirsk (Central Siberian Botanical Garden of SB RAS), and Moscow (N.V. Tsitsin Main Botanical Garden). Unfortunately, the assortment of shadbush varieties in Russia is still quite limited and is represented by only two varieties. In recent years, the development of methods of shadbush clonal micropropagation has been underway, which makes it possible to obtain planting material on an industrial scale and accelerate the reproduction of rare varieties.

Biochemical and Molecular Characterization of Musa sp. Cultured in Temporary Immersion Bioreactor

The genus Musa sp. contains commercially important fleshy fruit-producing plants, including plantains and bananas, with a strong potential for providing food security and sources of revenue to farmers. Concerns with the quality of vegetative tissues along with the possibility of the transmission of phytopathogens makes the availability of healthy plantlets limited for farmers. Micropropagation of plantains offers an alternative to producing large numbers of plantlets. However, conventional methods of micropropagation have high production costs and are labor-intensive. Recently, the temporary immersion bioreactor (TIB) has emerged as an alternative to conventional micropropagation (CM) methods. Our work utilized SEM microscopy (scanning electron microscope) and molecular and biochemical tools (qRT-PCR and ICP-OES) to characterize and compare the morphological properties, elemental composition, and photosynthetic gene expression of plantains cultured on TIB. Additionally, morphological features of growth and propagation rates were analyzed to compare outputs obtained from TIB and CM. Results showed higher growth and multiplication rates for plantlets cultivated in TIB. Gene expression analysis of selected photosynthetic genes demonstrated high transcript abundance of phosphoenolpyruvate carboxylase (PEPC) in plantain tissues obtained by TIB. Elemental composition analysis showed higher content of iron in plantains grown in TIB, suggesting a potential correlation with PEPC expression. These results demonstrate that micropropagation of Musa sp. via the liquid medium in TIB is an efficient and low-cost approach in comparison with solid media in CM.

Enhancing Gerbera Micropropagation Efficiency and Genetic Fidelity through Cytokinin and Auxin Combination Strategies

The present investigation was carried out to assess the impact of 6-benzyladenine (BA) in conjunction with Indole-3-butyric acid (IBA) on gerbera explant establishment under micropropagation. Additionally, the effects of BA, either individually or in combination with Kinetin (KIN), on shoot proliferation in two Gerbera cultivars, namely Kormoran and Dolores was experimented too. Throughout the experiment, various morphological changes were documented occurring during these micropropagation phases and also monitored potential genetic alterations using SSR markers. The studies revealed that the combination of BA and IBA yielded exceptional results, achieving a 100% success rate in explant regeneration within the shortest time frame. Notably, when BA and IBA were applied at lower concentrations, the number of shoots generated was reduced. However, the most substantial proliferation of shoots was observed when the growth medium contained 4 mg of BA and 0.5 mg of IBA per litre. Furthermore, our investigation into genetic fidelity using SSR analysis revealed no detectable polymorphism between the mother plant and the micropropagated plantlets in both the Gerbera cultivars, affirming the reliability of the micropropagation method in preserving genetic consistency.