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Salvia officinalis, commonly known as sage, is highly valued for its medicinal and ornamental properties. In the present work, 12 native sage populations of north-west Greece were evaluated for eight ornamental traits. Populations from the locations of Aristi, Kefalovryso and Igoumenitsa were selected as the best performing and for their preservation and availability in the market, their asexual propagation was investigated by (a) shoot cutting and (b) in vitro techniques. Propagation by cuttings was investigated during the four seasons. Aristi exhibited the highest rooting (65%) in spring with a well-developed root system (4.7 root number and 5.0 cm length) by applying 0.5 g·L−1 Indole-3-butyric acid, potassium salt (K-IBA), established on perlite under a fog system. However, the rooting performance of Aristi spring cuttings was not affected by different substrates of peat:perlite (0:1, 1:1, 1:2 v/v) or rooting systems (mist, fog) tested. Furthermore, the in vitro propagation of the selected sage populations was investigated using shoot tips as explants. After successful disinfection, the effect of Murashige and Skoog (MS) medium in ten different combinations of Indole-3-acetic acid (IAA), 6-Benzylaminopurine (BAP) and Thidiazuron (TDZ) were tested on shoot multiplication. Aristi presented the highest number of newly formed shoots on MS9 (0.1 mg·L−1 IAA and 0.8 mg·L−1 TDZ) and MS5 (0.1 mg·L−1 IAA and 0.8 mg·L−1 BAP) (3.35 and 3.21 new shoots/explant, respectively) with the highest shoot length (2.23 cm and 3.2 cm) and unexpected spontaneous root formation (64%) at MS5. The rooting ability of Aristi microshoots was further investigated in order to enhance their response. Of the three rooting variants tested, optimal rooting formation (100%) was observed on 0.9 mg·L−1 IAA (R3) combined with successful acclimatization (100%). Aristi exceeded the other populations in both the tested propagation systems, thus holding a strong potential for its introduction in the market as a competitive ornamental variety. [ABSTRACT FROM AUTHOR]

Hemp (Cannabis sativa L.) is commonly grown for the medicinal secondary metabolites produced by pistillate inflorescences. Micropropagation is a valuable method of propagating hemp plants because of the aseptic process and the production of true-to-type propagules. The hemp cultivar TJ’s CBD was used for a series of experiments to compare media inputs and practices for the clonal micropropagation of hemp. For stage I, shoot tips harvested from stock plants that were grown in a growth chamber produced less endogenous contamination in newly established cultures than shoot tips harvested from the greenhouse. In addition, stage I disinfection treatments with 20%, 40%, and 60% bleach (7.5% sodium hypochlorite) for 10 minutes had no differences in surface contamination rates. All concentrations were able to clean explants equally, and no damage to the explants was observed. For stage II, there were no differences in growth and multiplication rate between shoot tip or nodal explants. In addition, no differences were observed between the gelling agent’s agar, agargellan, and gellan gum at standard rates. When basal nutrient formulations were compared at standard rates and with their respective vitamins, Murashige and Skoog, Linsmaier & Skoog, and Driver & Kuniyuki Walnut media were found to be superior to Lloyd & McCown Woody Plant Medium. Media pH levels of 4.0, 5.0, 5.8, 6.0, and 7.0 were compared, and no differences were observed in final fresh weights, shoot lengths, or quality ratings. The pH levels of 5.8, 6.0, and 7.0 generated a greater number of lateral nodes. Sucrose levels of 0%, 1.5%, 3.0%, 4.5%, and 6.0% (wt/vol) were also compared, with the 1.5% and 3.0% rates showing greater fresh weights, shoot lengths, and quality ratings. Growth room temperatures of 22, 24, 26, and 28 °C were compared, with temperatures of 28 and 26 °C generating greater fresh weights, shoot lengths, numbers of nodes, and quality ratings compared with cooler temperatures. The cytokinins 6-enzylaminopurine (BA), 6-(γ,γ-dimethylallylamino) purine (2iP), and thidiazuron (TDZ) were compared at 1.0, 5.0, and 10.0 μM concentrations. The 5.0-μM TDZ treatment generated greater fresh weights and numbers of lateral nodes; however, it also produced the shortest shoot lengths and lowest quality ratings. The 2iP treatments at 1.0 and 5.0 μM, and the BA treatment at 1.0 μM produced the greatest quality ratings. The 5.0-μM 2iP level was considered the best treatment for stage II multiplication based on high ratings, in addition to the greater final fresh weights, shoot lengths, and numbers of nodes that were produced. For stage III experiments, the auxins indole 3-butyric acid (IBA) and 1-naphthylacetic acid (NAA) were compared at concentrations of 0.25, 0.5, and 2.5 μM. Auxin treatments of 0.25 μM NAA, 0.5 μM NAA, and 2.5 μM IBA generated the greatest final shoot fresh weights, root fresh weights, and numbers of nodes. However, the 2.5-μM IBA treatment resulted in a higher overall rating. For stage IV, ex vitro rooting and acclimation trials compared a dome and an intermittent mist system, as well as treated the unrooted cuttings with an externally applied auxin. Acclimating with a dome produced greater shoot heights, fresh shoot weights, and overall ratings compared with the mist system. The auxin treatment mildly increased fresh root weight, but was not as important to acclimation success as the domed environment. It has been concluded that a micropropagation system that uses lower rates of sucrose, higher growing temperatures, and lower rates of the cytokinins BA and 2iP are optimal for the micropropagation of hemp. In addition, when acclimating hemp plants from tissue culture, an in vitro stage III can be bypassed and plants can be rooted ex vitro during stage IV acclimation with a dome with or without additional auxin treatments.

Abstract Background Codonopsis pilosula (Franch.) Nannf. is a medicinal plant traditionally used in China, Korea, and Japan to treat many diseases including poor gastrointestinal function, low immunity, gastric ulcers, and chronic gastritis. The increasing therapeutic and preventive use of C. pilosula has subsequently led to depletion of the natural populations of this species thus necessitating propagation of this important medicinal plant. Here, we developed an efficient and effective in vitro propagation protocol for C. pilosula using apical shoot segments. We tested various plant tissue culture media for the growth of C. pilosula and evaluated the effects of plant growth regulators on the shoot proliferation and rooting of regenerated C. pilosula plants. Furthermore, the tissues (roots and shoots) of maternal and in vitro-regenerated C. pilosula plants were subjected to Fourier-transform near-infrared (FT-NIR) spectrometry, Gas chromatography-mass spectrometry (GC–MS), and their total flavonoids, phenolics, and antioxidant capacity were determined and compared. Results Full-strength Murashige and Skoog (MS) medium augmented with vitamins and benzylaminopurine (1.5 mg·L−1) regenerated the highest shoot number (12 ± 0.46) per explant. MS medium augmented with indole-3-acetic acid (1.0 mg·L−1) produced the highest root number (9 ± 0.89) and maximum root length (20.88 ± 1.48 mm) from regenerated C. pilosula shoots. The survival rate of in vitro-regenerated C. pilosula plants was 94.00% after acclimatization. The maternal and in vitro-regenerated C. pilosula plant tissues showed similar FT-NIR spectra, total phenolics, total flavonoids, phytochemical composition, and antioxidant activity. Randomly amplified polymorphic DNA (RAPD) test confirmed the genetic fidelity of regenerated C. pilosula plants. Conclusions The proposed in vitro propagation protocol may be useful for the rapid mass multiplication and production of high quality C. pilosula as well as for germplasm preservation to ensure sustainable supply amidst the ever-increasing demand.

The article is a review of the book "Orchids phytochemistry, biology and horticulture: Fundamentals and applications" edited by Jean-Michel Merillon and Hippolyte Kodja. The book covers various aspects of orchids, including their biodiversity, biology, biotechnology, and uses in horticulture and medicine. It consists of 26 chapters divided into six parts, discussing topics such as the distribution and ecology of orchids, their symbiotic relationship with mycorrhizal fungi, horticultural practices, agri-food applications, phytochemistry, medicinal properties, and cosmetic applications. The book provides a comprehensive reference for researchers and scholars in various fields related to orchids. [Extracted from the article]

The article discusses a new cultivar of the Lycoris chinensis flower called Jin Ling Tian Cheng. Lycoris is a perennial bulbous flower known for its surprising blooms and diverse colors. The new cultivar was selected from the L. chinensis population and features orange-yellow flowers with wider petals and smoother edges compared to the typical yellow L. chinensis. The article also provides information on the origin and development of Lycoris breeding, propagation methods, cultivation tips, and potential applications in gardens and as cut flowers. The new cultivar is available through the Institute of Botany, Jiangsu Province and Chinese Academy of Sciences. [Extracted from the article]

The article discusses a new cultivar of the Lycoris longituba plant called 'Yi Xian Fen Zhuang'. Lycoris is a group of bulbous flowering plants native to Asia and known for its showy flowers. The new cultivar has pink-purple flowers, distinguishing it from the white flowers of L. longituba. The article provides information on the origin, description, propagation, cultivation, and availability of the new cultivar. It also mentions the research team's work in breeding new and superior strains of Lycoris with high ornamental value. [Extracted from the article]

Endosperm as a result of double fertilization in Angiospermae shows high level chromosomes and polyploidy. It is also considered as dead tissue that is unable to be generated to form plantlet. The aim of this research is to determine the effect of kinetin and 2.4-dichlorophenoxyacetic acid (2.4-D) in induction of callus formation of mountain papaya. This research used factorial randomized block design with eighteen groups, one fruit was used for one experimental group. Culture using Murashige and Skog (MS) media with combination of three level of kinetin [(0, 1, 3) mg L-1] and six level of [2.4-D (0, 1, 2, 3, 4, 5) mg L-1]. Concentration of 2 mg L-1 2.4-D was the best treatment in stimulating callus growth by producing the highest percentage of callus, 24.44 %. The combination of 1 mg L-1 kinetin and 3 mg L-1 2.4-D was the best treatment in accelerating callus induction of the endosperm was 19.84 d after planting. In this study shows that induction callus of endosperm of mountain papaya could be stimulates by kinetin and 2.4-D.

Eryngium foetidum L., a biennial herb with diverse applications in food and traditional medicine, holds economic and pharmacological significance. Given its growing commercial interest, implementing biotechnological approaches like plant tissue culture is vital for sustainable propagation and metabolite production. In this study, we investigated the in vitro photoautotrophic potential of Eryngium foetidum, examining growth, chlorophyll a fluorescence, photosynthetic pigments, and anatomical features under sucrose concentrations (0 and 30 g L−1) and gas exchange rate (14 and 25 μL L−1 s−1 CO2). Acclimatization and survival rates of plants after ex vitro transfer were also assessed. Eryngium foetidum exhibited robust growth in both photoautotrophic and photomixotrophic conditions, with natural ventilation significantly enhancing plant development. Chlorophyll a fluorescence and photosynthetic performance were influenced by sucrose and gas exchange, highlighting the importance of these factors in plant micropropagation. Moreover, the species demonstrated remarkable plasticity during acclimatization, with high survival rates and rapid inflorescence development. The research provides valuable insights into optimizing in vitro cultivation conditions for Eryngium foetidum, emphasizing the potential for large-scale clonal propagation and exploring secondary metabolites. The observed phenotypic plasticity underscores the adaptability of the species to diverse environments. These biotechnological strategies open avenues for future studies, including the application of elicitors for enhanced secondary metabolite production.

Kalanchoë gastonis-bonnieri Raym.-Hamet & Perrier is a plant used for medicinal purposes in the treatment of several ailments. The aim of this study was to analyze the chemical profile of extracts from K. gastonis-bonnieri embryogenic calli, generated from genetically transformed roots by Agrobacterium rhizogenes. Putative transformants were verified by PCR. Hydroalcoholic extracts were obtained and the chemical profile was analyzed by LC-ESI-MS/MS. Root formation was obtained from 80% of infected seedlings. Fifteen root lines were isolated, and two lines showed prominent longitudinal growth and profuse branching in the B5 semi-solid medium. In all lines, the formation of nodules and later embryogenic callus was observed. Putative transgenic root lines were cultivated in free-plant growth regulators B5 medium. In the two selected lines, the PCR amplification of rolA, rolB, rolC, rolD, and aux1 genes was detected. The extract of embryogenic calli showed 60 chemical compounds tentatively identified, such as ferulic acid, quinic acid, neobaisoflavone, and malic acid, among others, and the chemical profile was different in comparison to wild-type extracts. This is the first study reporting the analysis of the chemical profile of hairy root extracts derived from Kalanchoë gastonis-bonnieri. This work displays the great potential for obtaining chemical compounds of pharmacological importance from hairy roots and facilitates the identification of new useful drugs against human chronic-degenerative diseases.

CRISPR/Cas9 systems are commonly used for plant genome editing; however, the generation of homozygous mutant lines in Medicago truncatula remains challenging. Here, we present a CRISPR/Cas9-based protocol that allows the efficient generation of M. truncatula mutants. Gene editing was performed for the LysM receptor kinase gene MtLYK10 and two major facilitator superfamily transporter genes. The functionality of CRISPR/Cas9 vectors was tested in Nicotiana benthamiana leaves by editing a co-transformed GUSPlus gene. Transformed M. truncatula leaf explants were regenerated to whole plants at high efficiency (80%). An editing efficiency (frequency of mutations at a given target site) of up to 70% was reached in the regenerated plants. Plants with MtLYK10 knockout mutations were propagated, and three independent homozygous mutant lines were further characterized. No off-target mutations were identified in these lyk10 mutants. Finally, the lyk10 mutants and wild-type plants were compared with respect to the formation of root nodules induced by nitrogen-fixing Sinorhizobium meliloti bacteria. Nodule formation was considerably delayed in the three lyk10 mutant lines. Surprisingly, the size of the rare nodules in mutant plants was higher than in wild-type plants. In conclusion, the symbiotic characterization of lyk10 mutants generated with the developed CRISPR/Cas9 protocol indicated a role of MtLYK10 in nodule formation.

This document discusses the importance of plant secondary metabolites in the communication and defense mechanisms of plants. These metabolites also have potential applications in human health due to their antioxidant and toxin-chelating properties. The document introduces a special issue of the journal Plants focused on plant tissue culture and secondary metabolite production. It includes research papers on various plant species, such as Ruta chalepensis, Stevia rebaudiana, Curcuma longa, Hypericum species, Taraxacum officinale, and Sideritis scardica, highlighting their potential for secondary metabolite production and their physiological characteristics. The authors of the document have contributed equally to the article and declare no conflicts of interest. [Extracted from the article]

The myrtle (Myrtus communis) plant naturally grows in the temperate Mediterranean and subtropical regions and is used for various purposes; thus, it is among the promising species of horticultural crops. This study aimed to evaluate and compare the performance of different propagation systems, including rooting, solid media propagation, rooting, and with the Plantform bioreactor system, in achieving healthy and rapid growth of four myrtle genotypes with diverse genetic origins and well-regional adaptation. The selection of myrtle genotypes with distinct genetic backgrounds and proven adaptability to specific regions allowed for a comprehensive assessment of the propagation systems under investigation. Present findings proved that the Plantform system, the new-generation tissue culture system, was quite successful in micropropagation and rooting myrtle genotypes. We succeeded in vitro micropropagation and rooting of diverse wild myrtle genotypes, enabling year-round propagation without reliance on specific seasons or environmental conditions. The process involved initiating cultures from explants and multiplying them through shoot proliferation in a controlled environment. This contributes to sustainable plant propagation, preserving and utilizing genetic resources for conservation and agriculture.

Cannabis sativa L. (hemp) has a global distribution and social impact, and it is widely used as a medicinal plant, food ingredient, and textile fiber. Its roots have received less attention than other parts, especially the inflorescence, leaves, and shoots. Triterpenoids, including friedelin and epifriedelanol, have been found in hemp roots, and their anti-inflammatory effects have been reported. In this study, the potential enhancement of triterpenoid accumulation in the roots of C. sativa by elicitation was examined. Hairy roots were successfully established, and they contained 2.02-fold higher triterpenoid levels than natural roots. Furthermore, hairy roots treated with 75 μM salicylic acid had 1.95-fold higher friedelin levels (0.963 mg/g DW) and 1.4-fold higher epifriedelanol levels (0.685 mg/g DW) than untreated hairy roots. These results suggested that the elucidation of hairy root cultures using an optimized elicitor could represent an alternative strategy to produce the valuable triterpenoids friedelin and epifriedelanol.

Background: Gossypium hirsutum undergoes rapid clonal propagation to regenerate a mature plant through tissue culture. However, the correlation between cotton leaf regeneration, callus induction, and regeneration ability was still obscure. In this research, cotton leaf regeneration level for 21 accessions in the field (new leaves) was observed after the first harvest, and a comparison between field regeneration level and callus induction with its regeneration capacity (new shoots and roots) for the same 21 accessions was carried out. Agronomic traits, including plant height, leaf area, fresh leaf weight, dry leaf weight, the number of flowers and bolls, and biochemical (proline content) and physiological (chlorophyll and carotenoid content) traits during the flowering stage of 21 upland cotton accessions, were investigated. Result: A significant correlation between physiological parameters and callus induction was discovered. Callus induction and regeneration capacity of roots and shoots for hypocotyl, cotyledons, and shoot tip tissues were used to validate field leaf regeneration level after the first harvest. CCRI 24 showed significant leaf regeneration in the field and callus induction capacity through callus induction and regeneration. Conclusion: We found a substantial relationship between field regeneration capability and callus induction with its regeneration capacity for the hypocotyl, cotyledons, and shoot tip. The results showed that ZS061, Lumian 378, Jimian 863, and ZS065 have the highest moisture retention capacity, while CCRI 24, Liaoyang Duomaomian, and Beizhe Gongshemian have the lowest moisture retention capacity. CCRI 24 has the highest leaf regeneration capacity in the field, while Beizhe Gongshemian has the lowest leaf regeneration capacity. All our result provides a clue for checking the regeneration capacity through leaf regeneration level in the field. [ABSTRACT FROM AUTHOR]

Inula crithmoides L. (golden samphire) is an edible aromatic halophyte species with confirmed nutritional and medicinal properties attributed to the presence of important metabolites, including proteins, carotenoids, vitamins, and minerals. Therefore, this study aimed at establishing a micropropagation protocol for golden samphire that can serve as a nursery approach to its standardized commercial cultivation. For that purpose, a complete regeneration protocol was developed by improving shoot multiplication from nodal explants, rooting, and acclimatization methodologies. The treatment with BAP alone induced the maximum shoot formation (7–7.8 shoots/explant), while IAA treatment increased the shoot height (9.26–9.5 cm). Furthermore, the treatment that coupled best shoot multiplication (7.8 shoots/explant) and highest shoot height (7.58 cm) was MS medium supplemented with 0.25 mg/L BAP. Moreover, all shoots produced roots (100% rooting), and multiplication treatments did not exert significant effect on root length (7.8–9.7 cm/plantlet). Moreover, by the end of the rooting phase, plantlets cultivated with 0.25 mg/L BAP had the highest shoot number (4.2 shoots/plantlet), and plantlets from 0.6 mg/L IAA + 1 mg/L BAP presented the highest shoot height (14.2 cm) similar to control plantlets (14.0 cm). The survival up to the ex-vitro acclimatization stage was increased from 9.8% (control) to 83.3%, when plants were treated with a paraffin solution. Nevertheless, the in vitro multiplication of golden samphire is a promising way for its rapid propagation and can be used as a nursery method, contributing to the development of this species as an alternative food and medicinal crop. [ABSTRACT FROM AUTHOR]

Nowadays use of aquatic plants for aquarium ornamentation as well as maintaining water quality is getting increasing interest. To fulfil the ever-growing demand in these areas, there is a need for non-conventional methods of massive and cost-effective clonal propagation of such plants. The present work was carried out with the purpose to set up a commercial in vitro micropropagation for three aquatic plant species (Lobelia cardinalis, Staurogyne repens, and Alternanthera reineckii). The explants were successfully surface sterilized and cultured in the solid Murashige and Skoog culture medium supplemented with 30 g L−1 sucrose, 5 g L−1 agar, and different combinations and concentrations of plant growth regulators (kinetin: 1, 2, and 3 mg L−1 and Indole-3-acetic acid [IAA]: 0, 0.2, and 0.5 mg L−1). Moreover, the effects of different explant types (shoot tip and nodal segments) were investigated on the multiplication rate as well as growth-related parameters such as plant length, number of roots, and root length. According to our results, the nodal explants were superior to shoot tip for micro shoot induction and leaflet production in three aquatic species. Both shoot and root induction were successfully achieved in the same medium, resulting in the plantlet regeneration through a single-step protocol without the need to further costly and time-consuming sub-culturing activities. Although the MS medium containing 2 mg L−1 kinetin gave the highest proliferation rate in three aquatic plants, however, there were some differences in response of each species to the IAA levels, highlighting the importance of specific behavior of each species. In general, enrichment of culture media with trace amount of IAA (0.2 and 0.5 mg L−1) significantly improved the proliferation rate and subsequent plantlet regeneration. The hardening and acclimatization of in vitro raised plantlets were successfully undertaken with 100% survival rate in water containing culture trays under maintaining of overwater relative humidity of 85% in greenhouse conditions. Altogether, the procedure presented here has great commercialization potential for rapid and large-scale micropropagation of these aquatic species especially in aquarium industries. [ABSTRACT FROM AUTHOR]

This study explored the possible role of magnesium nanoparticles (nMg) on plant growth, metabolism, and gene regulation in pepper. The physiological and molecular responses of pepper plants in presence of different concentrations of nMg (75, 225, and 675 mg/L) or its bulk analog (bMg) in the culture medium were investigated. We focused on how supplementing the culture medium with nMg affected biomass accumulation, activities of several key enzymes (catalase, peroxidase, and phenylalanine ammonia-lyase (PAL)), and expression of two vital transcription factors, WRKY1 and bZIP. Our results demonstrate that nMg and bMg improved biomass accumulation in shoots and roots. The nMg treatments up-regulated the expression of two transcription factors, WRKY1 and bZIP, which are transcriptionally involved in regulating defense/stress responses and secondary metabolism. The nanomaterials enhanced the activity of nitrate reductase, a nitrogen assimilation index. The nano-supplement efficiently stimulated the activities of catalase and peroxidase. The applied treatments also induced the PAL activity, which catalyzes a checkpoint reaction in the biosynthesis of phenylpropanoids. The supplementation of culture medium with nMg augmented the concentrations of proline and phenols. This study suggests the potential contributions of WRKY1 and bZIP transcription factors in regulating plant responses to nanomaterials. Nanomaterials, such as nMg, may improve the efficiency of plant tissue cultures. [ABSTRACT FROM AUTHOR]

• The newly developed three temporary immersion bioreactors showed superior response in multiple shoot induction and the plantlets showed better survival under ex vitro conditions than the semisolid culture system. • The bioreactor grown plants showed superior photosynthetic functions. • The flow cytometry and simple-sequence repeat markers analysis showed genetic integrity of the plantlets grown in TIBs. Temporary immersion bioreactor (TIB) technology offers a cost-effective means of in vitro micropropagation for large-scale plantlet production, and is increasingly being recognized as a valuable tool in the plant tissue culture industry. However, the limited accessibility of the bioreactor in certain markets is hindering its potential benefits. Here, we report three different TIB systems for large-scale plantlet production in banana. Shoot tip explants of commercially important banana cultivars such as Grand Naine (AAA), Red Banana (AAA), Nendran (AAB), and Ney Poovan (AB) were initiated aseptically in semisolid medium and cultured in the TIB systems using liquid medium supplemented with 4.0 mg/l 6- benzylaminopurine (BAP), 0.2 mg/l indole-3-acetic acid (IAA), and 30 g/l sucrose for shoot multiplication and liquid medium supplemented with 1.0 mg/l indole-3-butyric acid (IBA) and 2.0 mg/l 1-naphthaleneacetic acid (NAA) for rooting of shoots. The liquid media were supplied periodically (2 min for every 6-hr frequency) to the growth vessels and recorded the data on number of shoots per explant, shoot length and number of roots per shoot. The data were compared with explants cultured in semisolid medium. The three TIBs showed superior response over semisolid culture system in shoots per explant, shoot length and number of roots per shoot. There was no significant difference within TIBs in photosynthetic efficiency as measured by chlorophyll pigment analysis. Plantlets cultured in TIBs showed better survival under ex vitro conditions. The genetic integrity of the plantlets grown in TIBs were confirmed by flow cytometry and simple-sequence repeat (SSR) markers analysis. The developed TIBs and protocol can be utilized for commercial purpose. [ABSTRACT FROM AUTHOR]

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Catharanthus roseus L. (G.) Don is the most widely studied plant because of its high pharmacological value. In vitro culture uses various plant parts such as leaves, nodes, internodes and roots for inducing callus and subsequent plant regeneration in C. roseus. However, till now, little work has been conducted on anther tissue using plant tissue culture techniques. Therefore, the aim of this work is to establish a protocol for in vitro induction of callus by utilizing anthers as explants in MS (Murashige and Skoog) medium fortified with different concentrations and combinations of PGRs. The best callusing medium contains high α-naphthalene acetic acid (NAA) and low kinetin (Kn) concentrations showing a callusing frequency of 86.6%. SEM–EDX analysis was carried out to compare the elemental distribution on the surfaces of anther and anther-derived calli, and the two were noted to be nearly identical in their elemental composition. Gas chromatography–mass spectrometry (GC–MS) analysis of methanol extracts of anther and anther-derived calli was conducted, which revealed the presence of a wide range of phytocompounds. Some of them are ajmalicine, vindolinine, coronaridine, squalene, pleiocarpamine, stigmasterol, etc. More importantly, about 17 compounds are exclusively present in anther-derived callus (not in anther) of Catharanthus. The ploidy status of anther-derived callus was examined via flow cytometry (FCM), and it was estimated to be 0.76 pg, showing the haploid nature of callus. The present work therefore represents an efficient way to produce high-value medicinal compounds from anther callus in a lesser period of time on a larger scale. [ABSTRACT FROM AUTHOR]

In recent years, light emitting diodes (LEDs), due to their low energy consumption, low heat emission and specific wavelength irradiation, have become an alternative to fluorescent lamps (FLs) in plant tissue culture. The aim of this study was to investigate the effects of various LED light sources on the in vitro growth and rooting of plum rootstock Saint Julien (Prunus domestica subsp. insititia). The test plantlets were cultivated under a Philips GreenPower LEDs research module illumination system with four spectral regions: white (W), red (R), blue (B) and mixed (W:R:B:far-red = 1:1:1:1). The control plantlets were cultivated under fluorescent lamps (FL) and the photosynthetic photon flux density (PPFD) of all treatments was set at 87 ± 7.5 μmol m−2 s−1. The effect of light source on the selected physiological, biochemical and growth parameters of plantlets was monitored. Additionally, microscopic observations of leaf anatomy, leaf morphometric parameters and stomata characteristics were carried out. The results showed that the multiplication index (MI) varied from 8.3 (B) to 16.3 (R). The MI of plantlets grown under mixed light (WBR) was 9, lower compared to the control (FL) and white light (W), being 12.7 and 10.7, respectively. In addition, a mixed light (WBR) favored plantlets' stem growth and biomass accumulation at the multiplication stage. Considering these three indicators, we could conclude that under the mixed light, the microplants were of better quality and therefore mixed light (WBR) was more suitable during the multiplication phase. A reduction in both net photosynthesis rate and stomatal conductance in the leaves of plants grown under B were observed. The quantum yield (Yield = FV/FM), which represents the potential photochemical activity of PS II, ranged from 0.805 to 0.831 and corresponded to the typical photochemical activity (0.750–0.830) in the leaves of unstressed healthy plants. The red light had a beneficial effect on the rooting of plum plants; the rooting was over 98%, significantly higher than for the control (FL, 68%) and the mixed light (WBR, 19%). In conclusion, the mixed light (WBR) turned out to be the best choice during the multiplication phase and the red LED light was more suitable during the rooting stage. [ABSTRACT FROM AUTHOR]

Application of chemical mutagens is used for artificially induced in vitro mutation to develop new cultivars with elite characteristics. However, the optimization of selecting proper mutagen, its concentration, and exposure time is of utmost importance, especially for plants containing noteworthy secondary metabolites. In this study, the effect of sodium azide (NaN3) and ethyl methanesulfonate (EMS) in different concentrations (0.025, 0.05, 0.1, and 0.2 mg l−1), and treatment time (30, 60, and 120 min) was investigated on Bacopa monnieri; an important medicinal plant. The maximum shoot counts (57.0) were achieved from the combination of 0.10 mg l−1 EMS × 60 min. Whereas, maximum shoot length (4.07 cm), node numbers (4.97) and leaf numbers (12,23) were achieved from the combination of 0.20 mg l−1 EMS × 120 min, respectively. Combination of 0.025 mg l−1 NaN3 × 120 mg/l yielded maximum shoot counts (52.30), shoot length (3.23 cm), node numbers (6.07) and leaf numbers (12.13). The trained model to predict the outputs were designed and calibrated with machine learning (ML) algorithms. Support Vector Classifier (SVC), Gaussian Process (GP), Extreme Gradient Boosting (XGBoost), Random Forest (RF) models, and Multilayer Perceptron (MLP) neural network algorithms were used to discover the best models and their hyperparameters. The RF model gave exceptional results in the prediction of the outputs. F1 scores of the RF were acquired in the range of 0.98–1.00 for different outputs. The other models' F1 scores varied in the range of 0.65 and 0.85. The present work opens the new era of applying ML and artificial neural network (ANN) models in plant tissue culture with the possibility of application for other economic crops. [ABSTRACT FROM AUTHOR]

For many years, there has been a growing trend toward producing plants using tissue culture, the most efficient method at present. Every year, more and more protocols for micropropagation of economically valuable species are appearing. Many factors influence the regenerating explants under sterile laboratory conditions. One of the most important is light. The aim of the present study was to increase the efficiency of micropropagation of hellebore 'Molly's White' using energy-efficient light-emitting diodes (LEDs), which were compared to traditionally used fluorescent lamps (FLs). To choose the best light and reduce production costs, white, blue or red LEDs with two levels of photosynthetic photon flux density (PPFD), 40 and 70 μmol/m2/s, were used at the multiplication and rooting stages. LED light color has been shown to affect regeneration rate and plant growth in length during micropropagation, while both light parameters (color as well as intensity) affect the length of regenerating shoots and the content of assimilation pigments in plants. The use of white LED light, which gives the highest multiplication rate, at an intensity of 70 μmol/m2/s saves more than 57 kWh during an 8-week micropropagation cycle compared to conventional fluorescent lamps with the same parameters. [ABSTRACT FROM AUTHOR]

An increased dependency on plant-based resources for food, shelter, and medicinal usage has increased their sustainable and unsustainable exploitation. To use this resource sustainably, plant tissue culture (PTC) is one important technology. Among different PTC techniques, thin cell layer (TCL) technology is a relatively simple and easily adaptable technique for in vitro cultures of plants. This technique uses small explants about 0.5–2 mm in thickness excised from different plant organs. It has been successfully used in the large-scale propagation of vegetables, legumes, and plants with medicinal benefits. TCL technology has proven to be effective in stimulating various organogenic responses when combined with various new methods such as nanotechnology or microtome-based explantation, especially in tree species. It is considered an important tool in plant biotechnology. Although the morphogenetic response per explant is usually higher in conventional explants, the appropriate use of plant growth regulators and geometric factors in TCL has the potential to make it more efficient and beneficial. This article provides an overview of the concept of TCL as applied to different plant species, particularly trees, since there are few, if any, summaries of TCL technology, especially in trees. This review will certainly revitalize this important technology so that it can be used effectively for successful mass propagation in the field of plant tissue culture. [ABSTRACT FROM AUTHOR]

This study describes the in vitro propagation and ex vitro acclimatization of Philodendron erubescens pink princess, one of the most popular ornamental variegated foliage plants. For shoot proliferation, the protocorm-like bodies of the Philodendron pink princess were cultured on solid Murashige and Skoog (MS) media supplemented with 6-benzylaminopurine (BAP) and 1-naphthaleneacetic acid (NAA) at different concentrations. The results revealed that supplementation with BAP alone at a concentration of 1.0 mg/L yielded the maximum number of shoots and leaves. Furthermore, the application of BAP at 1.0 mg/L significantly enhanced the shoot proliferation of Philodendron pink princess when grown in liquid MS medium, yielding 11.2 shoots/explant and 4.7 leaves/explant. When the established microshoots were subjected to root induction using solid MS media supplemented with different kinds and concentrations of auxins, indole-3-butyric acid (IBA) at 3 mg/L resulted in the highest number of roots (3.2 roots/explant) and longest root length (1.9 cm). Three supporting materials, i.e., peat moss, vermiculite, and perlite, were used as planting media for the ex vitro acclimatization of the Philodendron pink princess plantlets. The results demonstrated that the in vitro plantlets acclimatized and exhibited a relatively high survival frequency in all planting media without morphological abnormalities. Peat moss outperformed all other types of planting media in terms of sustaining the vegetative growth of the plantlets. In the future, the approach established in this study could be employed for the extensive production of Philodendron pink princess. [ABSTRACT FROM AUTHOR]

Humankind has been unknowingly utilizing food fermentations since the first creation of bread, cheese, and other basic foods. Since the beginning of the last century, microbial fermentation has been extensively utilized for production of commodity chemicals. It has also gained substantial interest in recent decades due to its underlying applications in the preparation of natural and safe food ingredients including enzymes, antimicrobial agents, vitamins, organic acids, sweeteners, stabilizers, emulsifiers, oligosaccharides, amino acids, and thickening agents. In addition, some novel food ingredients that were conventionally made from some other sources such as plant tissue cultures or animals are now being introduced in the industry as 'fermentation products.' Some examples of such novel fermentation food ingredients include flavonoids, cultured meat products, food colorants, antioxidants, lipids, and fatty acids. This review summarizes some of the most prominent food ingredients and novel fermentation food products currently being produced via microbial fermentation as well as the strategies to enhance such fermentation processes. Additionally, economical feedstocks are discussed with their potential to be converted into value-added products with the help of microbial fermentations. [ABSTRACT FROM AUTHOR]

Afgekia mahidolae is a rare plant species that possesses antioxidant, antimicrobial, and wound healing properties. This study aimed to establish the in vitro culture of A. mahidolae and investigate the effects of elicitors on their phenolic and flavonoid production, including the antioxidant activities. The established callus was prepared in the form of cell suspension cultures to determine the effect of elicitors. After elicitation for 3 days, A. mahidolae cell suspension cultures treated by 5 µM salicylic acid or 100 mg/L yeast extract exhibited significantly higher levels of total phenolic and total flavonoid content than untreated cultures, which correlated to the antioxidant activities. In addition, the profiles of phenolic and flavonoid compounds in the callus and intact leaves of A. mahidolae were determined by LC-MS, which showed different phytochemicals. The findings of this study may encourage more sustainable development of A. mahidolae cultivation. [ABSTRACT FROM AUTHOR]

Silybum marianum (L.) is a medicinally important herb known for its liver-protecting compound, silymarin, which is an isomeric mixture of flavonolignans and taxifolin. In this study, a protocol was designed for the application of plant growth regulators at various concentrations and combinations of coconut water. This was aimed at initiating callus and silymarin production using different types of explants (root, hypoc -otyls, cotyledonary leaf) of S. marianum. Free radical scavenging activity of silymarin was evaluated in the extracts of callus cultures of these explants. In comparison with other plant growth regulators and explants, root explants that were cultured on MS medium and were enriched with 5.0 M NAA + 2.5 M BAP + 10% coconut water produced the most friable callus and had the highest possible level of silymarin and free radical scavenging activity. NAA + BAP + coconut water was found to be the best hormonal combination for callus initiation and mass production. This research can aid in the current understanding of the influence of plant growth regulators and different types of explants on the production of valuable secondary metabolites in S. marianum via in vitro cultures. [ABSTRACT FROM AUTHOR]

The family Zingiberaceae, commonly known as the ginger family, is economically sound due to their huge consumption as medicine, spices, dyes, perfumeries, etc. Gingers are widely distributed throughout the world, and India is one of the richest places with significant biodiversity of ginger plants. India is having 22 genera and about 176 species of gingers on which the maximum diversity of the ginger family is observed in Northeast India containing 19 genera and about 88 species. The wide occurrence of this family in the heterogeneous eco-geographical regions promotes natural genetic variations in intra- and interspecific level that constitutes the enormous genetic resources of gingers in India. The increasing demand for gingers leads to overexploitation and extensive cannibalism from natural habitats. Furthermore, habitat destruction and land crisis are also principal concerns imposing threats to the wild gingers. Some of the members in this family even became rare, endangered, and critically endangered according to the International Union for Conservation of Nature (IUCN). Plant tissue culture is now indispensable for the large-scale production of genetically identical rare plant genotypes that have remarkable commercial applications. Different approaches to in vitro propagation are the best alternatives for the mass production and conservation of Indian gingers. The primary aim of the article focused mainly on different in vitro aspects including updated micropropagation status of medicinally important Indian gingers, a threatened status that ultimately helps in the conservation of indigenous germplasm. [ABSTRACT FROM AUTHOR]

The range of floral colors is determined by the type of plant pigment accumulated by the plant. Anthocyanins are the most common flavonoid pigments in angiosperms; they provide a wide range of visible colors from redmagenta to blue-purple, products of cyanidin and delphinidin biosynthesis, respectively. For the floriculture industry, floral color is one of the most important ornamental characteristics for the development of new commercial varieties; however, most plant species are restricted to a certain color spectrum, limited by their own genetics. In fact, many ornamental crops lack bluish varieties due to the lack of activity of essential biosynthetic enzymes for the accumulation of delphinidin. An example is the poinsettia (Euphorbia pulcherrima Willd. ex Klotzsch), the ornamental plant symbol of Christmas and native to Mexico. Its popularity is the result of the variety of colors displayed by its bracts, a kind of modified leaves that accumulate reddish pigments based mainly on cyanidin and, to a lesser extent, on pelargonidin. The commercial success of this plant lies in the development of new varieties and, although consumers like the typical red color, they are also looking for poinsettias with new and innovative colors. Previous research has demonstrated the possibility of manipulating flower color through metabolic engineering of the anthocyanin biosynthesis pathway and plant tissue culture in different ornamental plant species. For example, transgenic cultivars of flowers such as roses, carnations or chrysanthemums owe their attractive bluish colors to a high and exclusive accumulation of delphinidin. Here, we discuss the possibilities of genetic engineering of the anthocyanin biosynthetic pathway in E. pulcherrima through the introduction of one or more foreign delphinidin biosynthetic genes under the transcriptional control of a pathway-specific promoter, and the genome editing possibilities as an alternative tool to modify the color of the bracts. In addition, some other approaches such as the appropriate selection of the cultivars that presented the most suitable intracellular conditions to accumulate delphinidin, as well as the incorporation of genes encoding anthocyanin-modifying enzymes or transcription factors to favor the bluish pigmentation of the flowers are also revised. [ABSTRACT FROM AUTHOR]

Background: Finding natural products with anticancer activity is an effective strategy to fight this disease. In this respect, Lepidium sativum or garden cress (family Brassicaceae) has been widely used worldwide for its wide therapeutic application, including anticancer and chemoprotective agents. Plant tissue culture techniques hold great promise for natural product enhancement without any climatic boundaries. In this study, glucosinolates and petroleum ether fractions were isolated from in vitro cell cultures and used against different carcinoma cell lines to investigate their anticancer potential. Methods: In this study, callus cultures from leaf and root explants were initiated, cell suspension cultures were established, and cell growth and viability profiles were characterized. Different amino acids were added as precursors to the cell suspension cultures to enhance glucosinolates accumulation. Gas chromatography–mass spectrometric analysis (GC–MS) of glucosinolates and petroleum ether fractions was performed, and all fractions were tested against different carcinoma cell lines. Results: The findings clarified that the maximum callus initiation percentage was obtained in the medium containing 1.0 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D) + 1.0 mg/l kinetin (Kin) (C1). The viable cell number of cell suspension cultures from leaves and roots increased until it reached the maximum values on day 15. Adding tyrosine and methionine to the cell suspension cultures was the most influential and recorded high glucosinolate percentages. 1H-Cyclopenta (b) pyridine-3-carbonitrile-4,5,6,7-tetrahydro-2-methylthio-4-spirocyclohexane was the main glucosinolate compound found in tyrosine-treated leaf suspension (GLT). Fifteen compounds were detected in the petroleum ether fraction in both cell suspensions initiated from the leaf and root (OL and OR). The major compounds were benzene-1,3,5-trimethyl (12.99%) in root cell suspension (OR), and benzene-2-ethyl-1,4-dimethyl (10.66%) in leaf cell suspension (OL). All glucosinolate extracts demonstrated significant anticancer activity against the prostate (PC3), lung (A-549), colorectal (caco2), and liver (HepG2) cell lines. Glucosinolates extracted from leaf cell suspension (GL) were the most active on the hepatocellular carcinoma cell line (HepG2) among all remaining glucosinolate extracts. Treated hepatocellular carcinoma with an IC50 of GL extract (47.5 ug/ml) upregulates pro-apoptotic BAX and downregulates anti-apoptotic BCL2, which disrupts the BAX/BCL2 ratio, leading to activation of caspase 3 inside treated HepG2 cells. Conclusions: The anticancer action of the GL extract was validated by the cell cycle study of its glucosinolates, which successfully promoted apoptosis and reduced hepatocellular growth by causing S-phase arrest. [ABSTRACT FROM AUTHOR]

Continuous discovery of novel in vitro plant culture practices is always essential to promote better plant growth in the shortest possible cultivation period. An alternative approach to conventional micropropagation practice could be achieved through biotization by inoculating selected Plant Growth Promoting Rhizobacteria (PGPR) into the plant tissue culture materials (e.g., callus, embryogenic callus, and plantlets). Such biotization process often allows the selected PGPR to form a sustaining population with various stages of in vitro plant tissues. During the biotization process, plant tissue culture material imposes developmental and metabolic changes and enhances its tolerance to abiotic and biotic stresses, thereby reducing mortality in the acclimatization and pre-nursery stages. Understanding the mechanisms is, therefore crucial for gaining insights into in vitro plant-microbe interactions. Studies of biochemical activities and compound identifications are always essential to evaluate in vitro plant-microbe interactions. Given the importance of biotization in promoting in vitro plant material growth, this review aims to provide a brief overview of the in vitro oil palm plant-microbe symbiosis system. [ABSTRACT FROM AUTHOR]

The goal of the current study was to compare the chemical composition of the roots, shoots, and leaves of wild-growing Eryngium maritimum L., and of in vitro and in field-cultivated plants in Latvia. The essential oil yield obtained by hydrodistillation ranged from 0.14% to 0.54%, while analysis of the chemical composition using GC-MS revealed a total of 44 different volatiles, with differences in the types and amounts of volatiles between the leaves and roots. Using 96-well plate techniques, the concentration of total phenolic compounds, saponins, and sugars in the aqueous ethanolic extracts of E. maritimum were assessed, along with their capacity to scavenge stable DPPH radicals. Extracts from roots had a lower concentration of total phenolic compounds compared to those from the leaves of wild grown and cultivated plants but did not differ from in vitro shoots. Root, leaf, and shoot samples of the same genotype from different growth conditions had approximately the same concentration of total saponins, while total sugar concentrations were higher in the roots. The growth conditions had a significant effect on the concentration of total phenolic compounds and antiradical activity, with differences that were significant observed between plant aboveground and belowground parts. Analysis using UHPLC-ESI-q-TOF-MS revealed 63 compounds, with amino acids and hydroxycinnamic acid derivatives (such as chlorogenic and rosmarinic acid) being the major compound groups that significantly differed between plant growth conditions. We also demonstrated that rapid screening of volatile compounds in in vitro plants using headspace gas chromatography mass spectrometry analyses can predict the formation of marker compounds in the same mericlones grown in field conditions. These findings provide valuable insights into the chemical composition of E. maritimum and its potential for use in various applications. [ABSTRACT FROM AUTHOR]

One of the most exotic and ubiquitous plant species, the cultivation of orchids continues to gain momentum worldwide. The growing popularity of orchids and their cultivation worldwide is attributed to substantial progress in approaches like classical breeding, plant tissue culture and biotechnological interventions. In addition to ornamental value, orchids are extensively studied for their pharmacological properties and are widely used as food supplements, flavours, and medicine in different regions of the world. The unprecedented advances in whole-genome sequencing and omics technologies have significantly improved our knowledge of orchid biology, leading to translational success in the production of exotic varieties. With the emerging investigations into the cultivation and global trade popularity of orchids in the present era, this thematic article provides comprehensive insights into the existing and emerging trends in orchid cultivation, diverse ethnomedicinal uses, and multi-faceted applications, and the need for the legalisation of orchids for conservation and trade in the present era. In this way, advances in breeding and molecular approaches aim to significantly improve orchid cultivation and its socio-economic attributes. However, a detailed understanding of challenges in orchid conservation and implementation of domestic/global legislative guidelines are necessary for the protection of endangered species and their legalised trade across the globe. [ABSTRACT FROM AUTHOR]

Panax ginseng is a major medicinal crop with pharmaceutical efficacy derived from ginsenoside metabolites. Despite its genome information, the inefficiency of ginseng transformation hinders the study of the molecular mechanism of ginseng plant metabolism. Thus, this protocol aimed to develop an easy and efficient system for ginseng transformation. We established a transformation system using ginseng callus cultured in a liquid medium, which has a higher rate compared with cotyledon explant. In addition, Agrobacterium tumefaciens has been used for plant transformation. Compared with the LBA4404 strain, C58C1 was inappropriate for ginseng transformation using ginseng callus. We induced and maintained calli in liquid medium and cut them into small pieces before infection. After infection, we selected calli that survived from the selection media until identification of newly growing cells. In β-glucuronidase (GUS) assay, the expression of the GUS gene was observed in cells that were newly generated from explants. We treated calli with 0.05 M of MgSO4 before infection. After MgSO4 pre-treatment, the transformation efficiency of growing cells around infected callus was increased than non-treated control. Moreover, we constructed and introduced a visible reporter RUBY system to easily identify transformed cells. Using this system, we could identify the cells by a red color with naked eyes. Based on our transformation protocol, the success rate has increased to 77.27% in surviving lines during selection culture. This stable ginseng transformation could facilitate the overexpression and knockout of ginseng lines for functional or synthetic biological studies. Key message: We re-designed the ginseng transformation protocol using ginseng calli for explant material, and MgSO4 pretreatment was conducted before Agrobacterium infection. RUBY reporter was successfully introduced in ginseng. [ABSTRACT FROM AUTHOR]

Eruca sativa is known in traditional medicine for its therapeutic effects, while young plants are used as a salad or green food. Recently, the consumption of rocket has increased considerably, so it has become very important for breeders. Plant tissue culture provides a platform to overcome the problems in improving this species. In the present study, an efficient protocol for in vitro shoot regeneration and propagation of recalcitrant rocket variety Domaća rukola was studied. Murashige and Skoog (MS, 1962) medium containing 0.1 mg L−1 BA and frequent subculture over a period of three weeks proved to be optimal for shoot multiplication with a multiplication index of over 3 and only 8.72% of hyperhydrated shoots without necrosis. Different concentrations of 2,4-D, BA, or TDZ in combination with NAA, with or without the presence of AgNO3, were tested for de novo shoot organogenesis (DNSO) from seedling explants. The hypocotyl explants cultured on MS with a combination of TDZ1.0+NAA0.1+AgNO3 5.0 mg L−1 regenerated viable shoots with the highest rate (25.38%) and an average number of 2.18 shoots per regenerating explant. Somatic embryogenesis from immature zygotic embryos proved to be the best way to regenerate a recalcitrant rocket cultivar. The highest embryogenic efficiency was achieved in explants cultured on MS medium containing 1.0 mg L−1 2,4-D with a frequency of 76.64% and 5.13 mean number of regenerated somatic embryos per explant, which were further converted into normal plants. Additionally, in vitro-produced rocket shoots could serve as a possible promising source for the production of flavonoid kaempferol with proven antioxidant properties. [ABSTRACT FROM AUTHOR]

In vitro culture, ensuring rapid multiplication and production of plant material under aseptic conditions, represents an excellent tool for ex-situ conservation of tree species biodiversity and can be used for the conservation, among others, of endangered and rare crops. Among the Pyrus communis L. cultivars that have been abandoned over the years due to changed cultivation requirements, but which are still used today in breeding programs, there is the 'Decana d'inverno'. Pear is generally considered a recalcitrant species for in vitro propagation due to weak multiplication rate, hyperhydricity, and susceptibility to phenolic oxidation. Therefore, the use of natural substances like neem oil (although little explored) represents one of the options to improve the in vitro plant's tissue culture. In this context, the aim of the present work was to evaluate the effect of adding neem oil (0.1 and 0.5 m L L−1) to the growth substrate in order to optimise the in vitro culture of the ancient pear tree cultivar 'Decana d'inverno'. The neem oil addition resulted in an increase in the number of shoots produced especially at both concentrations used. On the contrary, an increase in length of proliferated shoots was observed only with the addition of 0.1 mL L−1. The neem oil addition did not affect the explants viability, fresh and dry weights. Therefore, the present study demonstrated for the first time the possibility of using neem oil to optimise the in vitro culture of an ancient pear tree cultivar. [ABSTRACT FROM AUTHOR]

Key message: Plant regulatory noncoding RNAs (ncRNAs) have emerged as key modulators of gene expression during callus induction. Their further study may promote the design of innovative plant tissue culture protocols. The use of plants by humans has recently taken on a new and expanding insight due to the advent of genetic engineering technologies. In this context, callus cultures have shown remarkable potential for synthesizing valuable biomolecules, crop improvement, plant micropropagation, and biodiversity preservation. A crucial stage in callus production is the conversion of somatic cells into totipotent cells; compelling evidence indicates that stress factors, transcriptional regulators, and plant hormones can trigger this biological event. Besides, posttranscriptional regulators of gene expression might be essential participants in callus induction. However, research related to the analysis of noncoding RNAs (ncRNAs) that modulate callogenesis and plant cell dedifferentiation in vitro is still at an early stage. During the last decade, some relevant studies have enlightened the fact that different classes of ncRNAs, such as microRNAs (miRNAs), small interfering RNAs (siRNAs), and long noncoding RNAs (lncRNAs) are implicated in plant cell dedifferentiation through regulating the expression levels of diverse gene targets. Hence, understanding the molecular relevance of these ncRNAs in the aforesaid biological processes might represent a promising source of new biotechnological approaches for callus culture and plant improvement. In this current work, we review the experimental evidence regarding the prospective roles of ncRNAs in callus induction and plant cell dedifferentiation to promote this field of study. [ABSTRACT FROM AUTHOR]

• Reports long term clonal fidelity validation of M. acuminata. • Determination of quercetin, p -coumaric acid and eugenol within various tissues using HPTLC. • Assessment of anti- bacterial activity of various plant parts against prominent strains. • Validation of genetic conformity using SCoT and CBDP-gene targeted markers. • Model approach for other terrestrial orchids. Malaxis acuminata D. Don [ = Crepidium acuminatum (D. Don) Szlach.] is one of the prized medicinal orchids with large-scale application in various nutraceutical and herbal preparations. However, due to paucity in the availability of plant materials, its usage is often restricted. In order to ensure continuous supply of raw material for the pharmaceutical and herbal industries, plant tissue culture plays a crucial role. In the present research, transverse thin cell layer (t -TCL) derived in vitro cultures of M. acuminata were initiated and maintained in Murashige and Skoog (MS) medium supplemented with 1.5 mg/l meta -topolin (m T), 5.0 mg/l chitosan, 1.5 mg/l indole-3- butyric acid (IBA) and 5.0 mg/l phloroglucinol (PG) for three years time period under culture room conditions by regular intermittent sub-culturing. However, the success of in vitro propagated plant bio-resources are often adversely impacted due to the occurrence of clonal variations which is considered as a serious limitation for the commercial utilization. In order to ascertain the clonal fidelity of the long term - propagated in vitro lines of M. acuminata , genetic homogeneity amongst the regenerated plantlets was accessed using two advanced gene targeted molecular markers namely start codon targeted (SCoT) and CAAT box derived polymorphism (CBDP). The plants were found to exhibit 93.34% clonal fidelity in comparison to their mother plant. Keeping in consideration the therapeutic attributes of M. acuminata , the phytochemical makeup of the wild and in vitro - derived plant parts was compared using high performance thin layer chromatography (HPTLC) technique. Furthermore, M. acuminata is considered to be one the most precious medicinal orchids having rich reserves of biomolecules which can be used against various pathogenic bacterial strains. The antibacterial activities of various tissue extracts of M. acuminata were also evaluated against the opportunistic bacterial strains viz. Escherichia coli (MTCC43), Salmonella typhimurium (MTCC733), Staphylococcus aureus (MTCC96) and Bacillus subtilis (MTCC121). The results of antibacterial assays demonstrated the therapeutic efficacy of the in vitro - derived plant parts at identical magnitude with that of their wild counterparts. The present research provides a holistic insight into the clonal stability of the long term - propagated in vitro plants of M. acuminata with promising antibacterial activity. This study can also serve as a model approach for the maintenance of genetically stable in vitro lines of therapeutically important endangered orchid species over a period of time. [ABSTRACT FROM AUTHOR]

Haloxylon ammodendron (C.A.Mey) Bunge is one of the important species of arid desert vegetation in China, and it is also an ideal material for studying plant stress resistance, which plays an extremely important role in protecting desert ecosystems and preventing land desertification. However, there are relatively few studies on the regeneration of the fusiform, and the genetic transformation system has not been successfully reported, which restricts the in-depth study of the molecular mechanism of shuttle propagation and stress resistance. In the present study, the seeds, hypocotyls, cotyledons, cotyledon nodes, terminal buds and fixed buds were used as explants, and a set of tissue culture and plant regeneration system was established by inducing adventitious buds, adventitious bud rooting and transplanting. The results showed that amongst different H. ammodendron explants, i.e., seeds, hypocotyls, cotyledons and cotyledon nodes, the last induced budding effect was better. The optimal medium for inducing clandine buds by cotyledon segment differentiation is 0.5 mg·L-1 NAA+0.5 mg·L-1 6-BA induced budding rate was high, reaching 61.90%, rooting medium was 1/2 MS+1 mg·L-1 NAA +1 mg·L-1 IBA + 1 mg·L-1 IAA with a rooting rate of 50%. The results of this study will provide a theoretical basis for the genetic transformation of H. ammodendron. [ABSTRACT FROM AUTHOR]

Porang (Amorphophallus muelleri Blume.) usually used to functional food raw materials, caused its high glucomant content. Limitation of conventional propagation both vegetative and generative affected to decreasing of porang production to fulfil market demand. To solve the problem, plant tissue culture technique was chosen to rapid propagate bulbils of porang. So, the aim of this research to determine the combination and concentration of Plant growth Regulators (PGRs), such as 6-Benzylaminopurine (BAP), Naphthaleneacetic Acid (NAA), and Thidiazuron (TDZ) for quality of callus, also growth of callus and shoots for rapid propagation of porang, cause PGRs are one of factors that affect the success of in vitro propagation. Experimental design of this research was completely randomized designed with statistical analysis using analysis of variance (Anova) one way. Difference combination and concentration of cytokinin and auxin as manipulation variables, which several parameters growth of callus, shoots, and roots was observed to indicate the rapid organogenic of porang. The optimal shoot induction medium was Murashige and Skoog (MS) media with 5.0 mg.L-1 BAP and 0.2 mg.L-1 NAA on which number of shoots and shoot height reached 15±1.8 and (7.2±1.86) cm. After shoot formation stage, the proper media for rooting stage was determined, which was Murashige and Skoog (MS) media with 1.0 mg.L-1 NAA resulted the rooting rate reached 100% and roots appear earlier. Optimal combination and concentration of PGRs on micropropagation of Amorphophallus muelleri Blume. from bulbils explant should be known to increase the production of porang, which can fulfil the market demand. [ABSTRACT FROM AUTHOR]

The irrational exploitation of native species of arid and semi-arid areas of Mexico, in particular the huizache (Vachellia spp.), created the need to carry out research work that contributes to the reforestation of these areas. Vachellia spp. pods have been characterized by their nutritional value and use in ruminants, in addition to the fact that generally the seeds present germination problems under natural conditions, due to their hard and impermeable cover that prevents the passage of water. This is a problem that can be solved through the use of micropropagation. This technology has made it possible to obtain thousands of disease-free seedlings with characteristics identical to the plant that gave them origin, and so far there is no extensive research on the subject that includes diversity of species of the genus Vachellia L., some of the few examples are the case of the efficient induction of callus of the species Vachellia macracantha, induction of shoots and multiplication of Acacia auriculiformis, in vitro propagation of Acacia chundra and the hybrid Acacia mangium and A. mangium x A. auriculiformis hybrids, among other methodologies. Scarification methods (physical, chemical and mechanical) interrupt the dormancy of the seeds and by placing them in a suitable culture medium (macro and micronutrients, sucrose, growth regulators), large productions have been obtained in a short time. The aim of this Bibliographic Review was to compile the most relevant results on plant tissue culture in species of the genus Vachellia L. [ABSTRACT FROM AUTHOR]

Silver nanoparticles (AgNPs) are novel compounds used as antimicrobial and antiviral agents. In addition, AgNPs have been used to improve the growth of different plants, as well as the in vitro multiplication of plant material. In this work the effect of AgNPs on in vitro growth of 'Canino' and 'Mirlo Rojo' cultivars, as well as the leaf ion composition, are studied. Different concentrations of AgNPs (0, 25, 50, 75 and 100 mg L−1) were added to two culture systems: semisolid medium with agar (SSM) in jars and liquid medium in temporary immersion system (TIS). Proliferation (number of shoots), shoot length, productivity (number of shoot × average length), leaf surface, fresh and dry weight were measured. Additionally, the silver and other ion accumulation in the leaves were evaluated by inductively coupled plasma optical emission spectroscopy (ICP-OES) analysis. The productivity of 'Canino' and 'Mirlo Rojo' decreased when increasing the concentration of AgNPs in the semisolid medium. However, the use of AgNPs in the TIS improved the proliferation and productivity of 'Canino' and Mirlo Rojo', increasing biomass production, and the concentration of nutrients in the plants, although these effects are genotype-dependent. TISs are the best system for introducing silver into shoots, the optimum concentration being 50 mg L−1 for 'Canino' and 75 mg L−1 for 'Mirlo Rojo'. Principal component analysis, considering all the analyzed ions along the treatments, separates samples in two clear groups related to the culture system used. The use of bioreactors with a liquid medium has improved the productivity of 'Canino' and 'Mirlo Rojo' in the proliferation stage, avoiding hyperhydration and other disorders. The amount of metallic silver that penetrates apricot plant tissues depends on the culture system, cultivar and concentration of AgNPs added to the culture medium. Silver ion accumulation measured in the shoots grown in the TIS was higher than in shoots micropropagated in a semisolid medium, where it is barely detectable. Furthermore, AgNPs had a beneficial effect on plants grown in TIS. However, AgNPs had a detrimental effect when added to a semisolid medium. [ABSTRACT FROM AUTHOR]

At present, most precious compounds are still obtained by plant cultivation such as ginsenosides, glycyrrhizic acid, and paclitaxel, which cannot be easily obtained by artificial synthesis. Plant tissue culture technology is the most commonly used biotechnology tool, which can be used for a variety of studies such as the production of natural compounds, functional gene research, plant micropropagation, plant breeding, and crop improvement. Tissue culture material is a basic and important part of this issue. The formation of different plant tissues and natural products is affected by growth conditions and endogenous substances. The accumulation of secondary metabolites are affected by plant tissue type, culture method, and environmental stress. Multi-domain technologies are developing rapidly, and they have made outstanding contributions to the application of plant tissue culture. The modes of action have their own characteristics, covering the whole process of plant tissue from the induction, culture, and production of natural secondary metabolites. This paper reviews the induction mechanism of different plant tissues and the application of multi-domain technologies such as artificial intelligence, biosensors, bioreactors, multi-omics monitoring, and nanomaterials in plant tissue culture and the production of secondary metabolites. This will help to improve the tissue culture technology of medicinal plants and increase the availability and the yield of natural metabolites. [ABSTRACT FROM AUTHOR]