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A number of species have developed detoxification strategies that allow them to accumulate high amounts of heavy metals in their tissues and grow despite a high heavy metal (HM) concentration in soils. Thus, they are considered hyperaccumulator or accumulator species, attractive for phytoremediation purposes. Families reported to contain a high number of hyperaccumulator species are Asteraceae, Euphorbiaceae, Rubiaceae, Fabaceae, Scrophulariaceae, Myrtaceae, Proteaceae, Caryophylaceae, and Tiliaceae. Indeed, the family Scrophulariaceae has a large extension, with Buddleja, Eremophila, Scrophularia, and Verbascum being some of the best-known genera containing species used for medicinal purposes. This review focuses on identifying the species used in folk medicine that belong to these four genera and can accumulate high concentrations of heavy metals (HMs). Additionally, the human health risks related to the consumption of HM hyperaccumulator species are discussed, as well as the feasibility of plant tissue culture to provide safe vegetal material for medicinal uses or to estimate the intrinsic potential of species for phytoremediation purposes. The following species have demonstrated the ability to accumulate HMs and grow in mine or HM-polluted soils, as well as use in traditional medicine: Buddleja asiatica, B. cordata, B. davidii, B. lindleyana, B. macrostachya, B. paniculata, B. scordioides, Eremophila latrobei, Scrophularia canina subsp. bicolor, S. ningpoensis, S. striata, Verbascum bombyciferum, V. olympicum, V. phrygium, and V. thapsus. In some of these species, antioxidant machinery has been proven to participate as a defense strategy against exposure to high HM concentrations. Consuming plants that accumulate high amounts of HMs has been described as a potential risk to human health. Plant tissue culture has also been established in a few of these species, whether to propagate or use them as a bioactive phytochemical source. Promoting safety and quality control related to the HM concentrations in the tissues of these plants used by populations for curative purposes is necessary. Further research must be carried out on several species for phytoremediation purposes. [ABSTRACT FROM AUTHOR]

Whiteflies (Bemisia tabaci) and the diseases they transmit are a major detriment to crop yields and a significant contributor to world hunger. The highly evolved interactions of host plant, phloem-feeding insect vector with endosymbionts and persistently transmitted virus represent a tremendous challenge for interdisciplinary study. Presented here is the establishment of a colony of axenic whiteflies on tissue-cultured plants. Efficient colony establishment was achieved by a surface sterilization of eggs laid on axenic phototrophically tissue-cultured plants. The transfer of emerging whiteflies through coupled tissue culture vessels to new axenic plants facilitates robust subculturing and produces hundreds of whitefly adults per month. Whitefly proliferation on more than two dozen plant species is shown as well as in vitro testing of whitefly preference for different plants. This novel multi-organism system provides the high-level of biocontainment required by Federal permitting to conduct virus transmission experiments. Axenic whitefly adults were able to acquire and transmit a begomovirus into tissue-cultured plants, indicating that culturable gut microorganisms are not required for virus transmission. The approach described enables a wide range of hypotheses regarding whitefly phytopathology without the expense, facilities, and contamination ambiguity associated with current approaches. [ABSTRACT FROM AUTHOR]

Melatonin was discovered in plants in 1995, and so-called phytomelatonin. Its characteristic qualities as an antioxidant molecule and with amphiphilic nature make it an excellent candidate to use in plant biotechnological techniques. In plant physiology, melatonin has demonstrated with multiple studies great possibilities of application in crop improvement in aspects such as germination, rooting, plant growth, leaf-senescence, floration, fruit-set, parthenocarpy, fruit ripening and cut flower conservation, among others. In this work, we reviewed many studies on the possible role of melatonin to improve the different plant tissue culture goals, among them: the role of melatonin in callus culture induction and its effects in plant biomass production; the possible action of melatonin as an inductor or elicitor of different secondary metabolite generation in in vitro cultures; the interesting possibilities of melatonin as a protective agent (safener) against abiotic stressors present in in vitro cultures; the potential of melatonin in regulating plant organogenesis, modulating shooting/rooting responses; at last, the studies on the relevant role of melatonin in plant material cryopreservation and the improvement of cell transformation using A. tumefaciens have been discussed. [ABSTRACT FROM AUTHOR]

Watermelon (Citrullus lanatus) is one of the five most consumed fresh fruits and is a commercially important horticulture crop. The consumer demand for seedless watermelons has considerably increased due to their superior taste and ease of consumption. Seedless watermelon plants are triploids hence plant tissue culture technology which is the cultivation of plant cells, tissues, or organs on specially formulated nutrient media can facilitate the rapid multiplication of seedless watermelon varieties. However, hyperhydricity has been reported as a frequent problem for in vitro growth and development of watermelon. Hence, the present study highlights the application of silver nanoparticles biosynthesized from Parthenium hysterophorus (PHAgNPs) for reversal of hyperhydricity and enhancing in vitro shoot production in watermelon. The biosynthesized PHAgNPs was characterized using UV–vis spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction analysis and Transmission electron microscopy. Cotyledonary nodes used as explant and cultured in three types of cytokinins (BAP, TDZ and KIN) became hyperhydric, losing their ability to regenerate at high cytokinin concentrations (> 1.5 mg/L) irrespective of the cytokinin type. The optimum concentration of BAP (1.0 mg/L) fortified with PHAgNPs (15.0 mg/L) exhibited an improved response of 91 % and induced a maximum of 32.33 shoots with a maximum shoot length of 5.66 cm after 6 weeks. The incorporation of PHAgNPs reduced the percentage of hyperhydricity from 29 % to 1.33 % in regenerated shoots, which was confirmed by the reduction in relative water content (RWC) from 93.33 % to 80.33. Silver ions reduced H 2 O 2 content by 50 % and increased photosynthetic pigments (chlorophyll a, b and carotenoids) by 50 %, which contributed to the formation of healthy regenerated shoots, with opened stomata compared to closed in hyperhydric shoots. Compared with those of hyperhydric shoots, the gene expression patterns of 1-aminocyclopropane-1-carboxylase synthase (ACS1) and 1-aminocyclopropane-1-carboxylic acid oxidase (ACO1) decreased after the retroversion of shoots to 15 mg/L PHAgNP medium. The relative gene expression profiles of ACS1 and ACO1 in hyperhydric shoots were 8.8- and 1.8-fold greater than those in normal shoots, respectively. Profuse root growth was observed in MS basal media supplemented with IBA (1.0 mg/L) and PHAgNPs (10.0 mg/L), which resulted in the greatest number of roots (12.33), with an average root length of 4.33 cm and a response of 89.33 %. The genotoxic effect of PHAgNPs evaluated using RAPD and SCoT molecular markers, showed that the addition of low levels of silver nanoparticles to the medium did not cause any genetic variation. The efficient regeneration protocol standardized in this study provides a solution to solve the problem of hyperhydricity in watermelon regeneration. [Display omitted] • Silver nanoparticles was biosynthesized from Parthenium hysterophorus (PHAgNPs) leaf extracts. • Maximum number of multiple shoots (32.33 shoots/explant) was obtained from MS media supplemented with BAP (1.0 mg/L) and PHAgNPs (15.0 mg/L). • BAP (1.0 mg/L) and PHAgNPs (15.0 mg/L) supplemented media significantly reduced the percentage of hyperhydricity to 1.33 %, in contrast to the control (29 %). • Silver ions reduced H 2 O 2 content by 50 % and increased photosynthetic pigments (chlorophyll a, b and carotenoids) by 50 %. • Down regulation of ethylene-related genes (ACS1 and ACO1) after the retroversion of shoots from 15 mg/L PHAgNPs medium. • Genotoxicity analysis (RAPD and SCoT molecular markers), confirmed that the in vitro regenerated plants were genetically uniform to the mother plant. [ABSTRACT FROM AUTHOR]

Plant tissue culture biotechnology offers a sustainable way to adapt to climate change by propagating disease-free plants with the desired traits. This study investigated stakeholders' agronomic, social and economic evaluation of tissue culture plants (TCPs). It unravels micro-level outcomes and indicators that determine TCPs evaluation through a systematic literature review. The 35 articles examined show an imbalance of studies in the three evaluation streams, with social studies on horticultural crops, especially bananas in developing countries, dominating. The results of the TCPs evaluation vary considerably between stakeholders. The main determinants of evaluation are TCPs attributes. TCPs are very knowledge-intensive and costly to establish and manage. Therefore, customising of TCPs to local conditions and closer collaboration between stakeholders are essential for successful implementation. Advisory services, social entrepreneurship and networks are needed to create knowledge-sharing platforms and ecosystems. While single-stream evaluations reveal TCPs superiority over conventionally propagated crops, it is suggested that an integrated assessment be undertaken to combine social, economic, and agronomic indicators and evaluate the technology simultaneously. [ABSTRACT FROM AUTHOR]

Climate change and biodiversity conservation are two of the most urgent challenges of the twenty-first century. Current global climate models indicate that climate-related events will continue to increase in frequency and intensity, leading to severe impacts on ecosystems, particularly plant diversity. Despite the 2015 Paris Agreement, efforts to limit global warming to 1.5 degrees Celsius and secure adequate climate financing remain unsatisfactory. In addition, the National Biodiversity Strategic Action Plans (NBSAPs), a core mechanism of the Convention on Biological Diversity (CBD), have faced issues in implementation at the national level. Many NBSAPs lack clear and measurable biodiversity targets, which limits their effectiveness. This review presents a comprehensive analysis of these urgent issues, highlighting the significant challenges and deficiencies in current climate and biodiversity conservation policies. It evaluates the effectiveness of the Kew conservation strategy as a model for protecting and conserving plant diversity. Furthermore, this review underscores the pivotal role of plant tissue culture (PTC) technology in achieving plant conservation targets within the post-2020 global biodiversity framework. This review strongly supports the post-2020 global biodiversity framework and the integration of PTC into global plant conservation strategies to meet the ambitious 30-by-30 targets. This review also advocates for the establishment of the Kew-Wide Mechanism (KWM) to bolster climate resilience, reduce anthropogenic impacts on plant diversity, revitalize global conservation efforts, and accelerate ecosystem restoration in the face of ongoing climate change. Proposed as a comprehensive approach to plant conservation, the KWM offers a strategic, innovative, and scalable solution as the global economy transitions toward decarbonization. [ABSTRACT FROM AUTHOR]

ABSTRACT Nanoparticles (NPs) are well‐known biostimulants in plant biotechnology, utilised to enhance the physical properties of plants and exhibit positive effects on them. The important key role is the most suitable type, effective dose and size of NP to be used in plant tissue culture systems. In this study, various concentrations of silver nanoparticles (AgNPs; 0, 25, 50, 75 and 100 μg L−1) were tested as elicitors in callus culture with the aim of enhancing secondary metabolite production in lemon balm (Melissa officinalis L.). According to the results obtained, callus formation rates have shown an increase in all applications compared to the control group. The highest callus formation, weight and diameter were observed in 50 μg L−1 application. In this application, the callus structure was compact and its colour was green. However, the aromatic compounds, neral and geranial increased significantly in 25 μg L−1 application. The maximum increase in phenolic compounds such as caffeic acid, chlorogenic acid, proto‐catechic acid, hesperidin and p‐coumaric acid was observed in the 75 μg L−1 AgNP and the highest increase in rosmarinic acid compound was determined in the 50 μg L−1 application. The study found that AgNP applications are an effective method for increasing the production of secondary metabolites in medicinal and aromatic plants, such as lemon balm, in vitro. [ABSTRACT FROM AUTHOR]

Plant tissue culture is extensively employed in plant functional genomics research and crop genetic improvement breeding. The callus induction ability is critical for utilizing Agrobacterium-mediated genetic transformation. In this study, we conducted a genome-wide association study (GWAS) utilizing 368 rice accessions to identify traits associated with callus induction rate (CIR), resulting in the identification of a total of 104 significant SNP loci. Integrated with gene function annotation and transcriptome analysis, 13 high-confidence candidate genes involved in auxin-related, CYC cyclins, and histone H3K9-specific methyltransferase were identified in significant loci. Furthermore, we also verified a candidate gene, Os05g0493500 (OsCycB1;5), and employed the CRISPR/Cas9 system to generate OsCycB1;5 knockout mutants in rice (Oryza sativa L.). The OscycB1;5 mutant displays significantly reduced callus induction and proliferation capacity, this result indicating OsCycB1;5 is required for the callus formation in rice. Overall, this study provides several reliable loci and high-confidence candidate genes that may significantly affect callus formation in rice. This information will offer valuable insights into the mechanisms underlying callus formation not only in rice but also in other plants. [ABSTRACT FROM AUTHOR]

To replace explosive nitrate-based chemicals in MS medium, this study developed a new, safer, and more cost-effective method using fertilizer-grade calcium ammonium nitrate and ammonium sulfate. This approach replaces ammonium nitrate and potassium nitrate, ensuring both safety and cost efficiency for sugarcane propagation. Six local sugarcane varieties—Zhongtang1 (ZT1), Zhongtang3 (ZT3), Zhongtang6 (ZT6), Guitang42 (GT42), Guitang44 (GT44), and Guiliu 07150 (GT07150)—were used. In the control group (Ck), nitrate ions (NO3−) were 39.28 mM, and ammonium ions (NH4+) were 20.49 mM, with a 2:1 ratio. In the treatment groups, the concentrations of nitrate ions (NO3−) and ammonium ions (NH4+) included treatment 1 (19.69 mM NO3− and 10.3 mM NH4+), treatment 2 (29.54 mM and 15.44 mM), treatment 3 (39.38 mM and 20.59 mM), treatment 4 (49.225 mM and 25.74 mM), treatment 5 (59.07 mM and 30.89 mM), and treatment 6 (68.915 mM and 36.03 mM), respectively, all with the same 2:1 ratio. Fifty bottles per treatment, with three replicates, were used for each sugarcane plantlets treatment. After five subcultures, the optimal ratio was determined by assessing morphological and physiological parameters, nitrogen levels, and SOD enzyme activity. The results indicated that treatment 3 (39.38 mM and 20.59 mM) and treatment 4 (49.225 mM and 25.74 mM) had the best morphological and physiological indicators. The optimal doses of calcium ammonium nitrate and ammonium sulfate were found in treatments 3 and 4, as well as in the control, with no significant difference among them. However, treatment 3, due to its lower dose, was more cost effective. To improve cost efficiency in practical production, it is recommended to use the lower concentration ratio of treatment 3 for plant tissue culture plantlets. [ABSTRACT FROM AUTHOR]

Cannabis has been cultivated for thousands of years for a variety of purposes, including fiber, seeds, oil, and medicinal compounds. The cannabis industry is growing rapidly because several countries have recently legalized the use of cannabis. In these countries, the industry related to cannabinoid ingredients such as cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC) is steadily increasing every year. High concentrations of cannabinoids are mainly produced in unfertilized female flowers. Maintaining plants with high cannabinoid content is essential for producing uniform substances in large quantities. This study established an in vitro micropropagation protocol that can maintain the mother plant characteristics of Cannabis sativa 'Cheungsam'. As a result of this experiment, the shoot tips and nodes of Cannabis sativa 'Cheungsam' at various concentrations (0, 0.25, 0.5, 1.0 mg/L) of 2iP, BA, and mT plant growth regulators (PGRs), and all concentrations of 2iP showed better results compared to two other hormones. However, the cut surfaces turned black, and excessive hyperhydricity occurred. Based on these symptoms, activated charcoal was added to the medium with the assumption that necrosis and hyperhydricity occur due to the accumulation of reactive oxygen species (ROS). When treated with 0.5 g/L charcoal, hyperhydricity was not overcome, and there was no difference compared to the control. As a new alternative, we divided the experiments into MS (Murashige and Skoog) and DKW (Driver and Kuniyuki Walnut) medium, which were commercially available. As a result, the rate of hyperhydricity was reduced, the cut surface did not turn black, and the growth conditions were also improved. Subsequently, ½ MS medium and ½ DKW medium were treated with various concentrations of IBA alone and with combinations of IBA and NAA for rooting. As a result, ½ DKW with IBA 0.5 mg/L showed the highest rooting rate and the best root condition for Cheungsam. After 4 weeks, when considering rooted plants with a height above 5 cm that were acclimatized, the acclimatization rate reached 100%. In conclusion, the Cannabis sativa 'Cheungsam' plants used in this study produced healthy shoots on DKW medium containing 1.0 mg/L 2iP and 0.5 mg/L of IBA in ½ DKW medium showed the best rooting rate. [ABSTRACT FROM AUTHOR]

Conventional in vitro propagation using semisolid Murashige and Skoog (MS) culture systems is costly, labor-intensive, and requires substantial space for large-scale plant production. This study investigated the application of a temporary immersion bioreactor (TIB) system for the micropropagation of the banana cultivar Kluai Numwa Pakchong 50, as a promising platform for economical commercial production. The cultivation parameters affecting plantlet multiplication, including plant growth regulator (PGR) use, explant density, and immersion frequency, were examined. Additionally, the ex vitro acclimatization of well-developed in vitro plantlets was also evaluated. Using liquid MS medium supplemented with 7.5 mg/L 6-benzylaminopurine (BAP) in the TIB system yielded significantly better results than the conventional semisolid MS control system, producing more shoots (5.60 shoots/explant) and leaves (2.80 leaves/explant) with longer shoot length (2.19 cm). Optimal conditions in the TIB system included an inoculum density of five explants per culture vessel and an immersion frequency of once every 6 or 8 h for 2 min. For root induction, 0.5 mg/L indole-3-butyric acid (IBA) proved more effective than 1-naphthaleneacetic acid (NAA). After 30 days of ex vitro acclimatization, plantlets regenerated from the TIB system demonstrated high survival rates, vegetative growth performance, and root formation efficiency comparable to those from the semisolid culture system. These findings establish the TIB system as a promising platform for the mass propagation of the Kluai Numwa Pakchong 50 banana. The protocol developed in this study could potentially be adapted for large-scale production of other banana varieties. [ABSTRACT FROM AUTHOR]

Plant tissue culture is the best option in producing planting material for cultivation programmes of Gyrinops walla. Perfection of in vitro rooting protocol for this species is the most decisive step in micropropagation. In the present study the effect of physical state of substrate, different concentrations of sucrose, IBA and NAA in media containing activated charcoal on in vitro rooting was investigated. In experiment-1, 0.2% (w/v) activated charcoal was added in to the ½ MS medium fortified with 30 or 40 g/L sucrose and varying concentrations of IBA or NAA (0, 0.1, 0.5, 1.0 and 2.0 mg/L). In experiment-2, filter paper-bridged liquid medium was used with 40 g/L sugar and varying concentrations of IBA or NAA (0, 0.1, 0.5, 1.0 and 2.0 mg/L). The treatments were arranged in completely randomized design with three replicates. Percentage of rooted microshoots and number of roots per shoot were recorded. The ½ MS semi-solid medium with 0.2% activated charcoal, 40 g/L sucrose and 0.1 mg/L NAA resulted in the significantly highest rooting percentage (50%)while the significantly highest number of roots (6.0) were noted in ½ MS charcoal added (0.2%) basal medium with 40 g/L sucrose and 0.5 mg/L NAA. Shoots supported with filter paper bridge on ½ MS liquid medium, supplemented with 0.5 mg/L IBA, had the highest rooting percentage (66.7%) and the highest mean number of roots per shoot (3.0) (P<0.05). Results confirmed that in vitro rooting of G. walla could be perfected by optimizing the culture medium and its constituents. [ABSTRACT FROM AUTHOR]

An efficient protocol to standardize the multiple shoot induction of plantlets from nodal explants of Physalis angulata L. using MS basal media with the combination of Cytokinin and auxin was studied in this research. Physalis angulata is a cosmopolitan plant found in tropical and sub tropical regions of the world. The plant has wide range of therapeutic abilities in fields like ethnobotany, phytochemistry, pharmacology, antimicrobial, anti-inflammatory, antioxidant, immunoregulatory, anti-diabetic, anticancerous and antilipidiamic function especially the compound Physalin. For large scale production and extraction of these functional compounds from the plant, a conservational technique namely plant tissue culture was employed in this study, where, node from elite mother plant is taken and inoculated in the MS medium supplemented with combinations of cytokinin (BAP+KIN) and auxin (NAA+IAA) in various range of concentration was used. The aseptic condition was maintained and data's were analysed by ANOVA. From the result it was observed that highest concentration provided best result of number and length of shoots and roots. From this study, it is evident that plant tissue culture plays a role in not only conservation of RET plants but also common plants and weeds with therapeutic ability that could be accessed for large scale production and also compound elucidation. [ABSTRACT FROM AUTHOR]

Abstract A number of species have developed detoxification strategies that allow them to accumulate high amounts of heavy metals in their tissues and grow despite a high heavy metal (HM) concentration in soils. Thus, they are considered hyperaccumulator or accumulator species, attractive for phytoremediation purposes. Families reported to contain a high number of hyperaccumulator species are Asteraceae, Euphorbiaceae, Rubiaceae, Fabaceae, Scrophulariaceae, Myrtaceae, Proteaceae, Caryophylaceae, and Tiliaceae. Indeed, the family Scrophulariaceae has a large extension, with Buddleja, Eremophila, Scrophularia, and Verbascum being some of the best-known genera containing species used for medicinal purposes. This review focuses on identifying the species used in folk medicine that belong to these four genera and can accumulate high concentrations of heavy metals (HMs). Additionally, the human health risks related to the consumption of HM hyperaccumulator species are discussed, as well as the feasibility of plant tissue culture to provide safe vegetal material for medicinal uses or to estimate the intrinsic potential of species for phytoremediation purposes. The following species have demonstrated the ability to accumulate HMs and grow in mine or HM-polluted soils, as well as use in traditional medicine: Buddleja asiatica, B. cordata, B. davidii, B. lindleyana, B. macrostachya, B. paniculata, B. scordioides, Eremophila latrobei, Scrophularia canina subsp. bicolor, S. ningpoensis, S. striata, Verbascum bombyciferum, V. olympicum, V. phrygium, and V. thapsus. In some of these species, antioxidant machinery has been proven to participate as a defense strategy against exposure to high HM concentrations. Consuming plants that accumulate high amounts of HMs has been described as a potential risk to human health. Plant tissue culture has also been established in a few of these species, whether to propagate or use them as a bioactive phytochemical source. Promoting safety and quality control related to the HM concentrations in the tissues of these plants used by populations for curative purposes is necessary. Further research must be carried out on several species for phytoremediation purposes. Graphical abstract

Tissue culture techniques were used to produce large amounts of bioactive compounds with medicinal potential, overcoming space and time constraints for cancer prevention. Rice callus suspension cultures (RCSC) and seed extracts prepared from aromatic rice varieties were used to evaluate the cytotoxic impact on human colon and lung cancer cell lines, as well as a normal control cell line, using Taxol as a positive control. RCSC and seed extracts from two Indian aromatic rice varieties were applied at different concentrations to treat the cancer cell lines and normal lung fibroblasts over varying time intervals. Apoptosis was assessed in 1:5 dilutions of the A549 and HT-29 cell lines treated with RCSC for 72 h, using propidium iodide staining and flow cytometry. RCSC showed a more potent cytotoxic effect than seed extracts with minimal effect on the normal cell line, in contrast to Taxol. Confocal microscopy and flow cytometry further confirmed the apoptotic effect of RCSC. Gas chromatography-mass spectrometry-based metabolic profiling identified metabolites involved in cytotoxicity and highlighted altered pathways. RCSC is proposed as an alternative source for the development of novel anticancer drugs with reduced side effects.

Curcuminoids are the major specialized metabolites in Curcuma species that are used as natural dyes and therapeutic agents with multifold biological activities. Curcuminoids are produced by field cultivation of plants; however, this method suffers from long growth periods and genotypic and environmental conditions. Therefore, research efforts have been made to procure curcuminoids from plant tissue cultures of Curcuma species, especially Curcuma longa, C. aromatica, and C. zedoaria. Cell, hairy root, shoot, and microrhizome cultures have been established to produce curcuminoids in vitro. Elicitation processes have been implemented to improve curcuminoid accumulation in plant tissue cultures. In this review, we present the research conducted on curcuminoid production in plant tissue cultures. In addition, efforts have been made to summarize the research accomplishments in the production of curcuminoids in heterologous systems. Curcuminoids produced from cell and organ cultures are used for food coloring, flavoring, preservation, and therapeutic purposes. [ABSTRACT FROM AUTHOR]

Gentians (Gentiana spp.) as floriculture crops are constantly exposed to several fungal and viral pathogens in the field. Among the fungal diseases afflicting gentian production, gentian sclerotial flower blight caused by Ciborinia gentianae incurs economic losses, as it affects flowers before and after harvest. Currently, preventive measures for this disease are limited, and no resistant cultivars have been reported. This is partly because of the lack of a reliable infection system that could promote research on this plant-fungus interaction. In this study, Gentiana plant tissue culture material was inoculated with C. gentianae culture filtrate. We successfully demonstrated non-ascospore-mediated infection of C. gentianae. Inoculation of individual hyphal structures present in the culture filtrate suggested that sclerotial primordia are the main agents of this infection. Interestingly, our results indicated that primary infection of C. gentianae occurs in petals rather than leaves, which enables systemic infection and therefore mirrors the fungus's infection strategy observed in the field. Moreover, we showed that (i) non-ascospore hyphal structures can also cause disease in flowers grown in the field, and (ii) ascosporic infection can also be observed using the in vitro system, opening possibilities for both practical and basic research aimed to combat gentian sclerotial flower blight disease. [ABSTRACT FROM AUTHOR]

Micropropagation of explants in vitro has potency to address propagule demands for promoting large-scale vanilla production. Plant growth regulators (i.e., cytokinin, auxin) are necessary for the plant micropropagation success. Objective of this study is to determine the shoot multiplication and root development of Vanilla planifolia under the influence of different concentrations and combinations of BA and IBA for micropropagation. Sixty stem nodal segments of Vanilla planifolia were cultured on MS medium supplemented with IBA (0, 0.5, or 2.0 mg/L) and combined with BA (0, 0.5, 1.0, or 2.0 mg/L). Shoot multiplication and root induction were measured after 60 days of culture. The results show that the MS medium with 1 mg/L IBA hindered shoot growth, while the medium containing 1 mg/L BA yielded the highest number or weight of shoots per explant. For the root development, supplementing the medium with 0.5 mg/L or 1 mg/L IBA improved root length or number of roots per explant, respectively. This research establishes a valuable approach for vanilla micropropagation by utilising low concentrations of plant growth regulators and a rapid protocol. This paves the way for significant advancements in large-scale commercial production. [ABSTRACT FROM AUTHOR]

Plant cell and tissue culture have been used as the alternative and potential renewable source for the production of valuable phytochemicals. Elicitation offers a reliable in vitro approach to produce or enhance potential phytochemicals. α-tocopherol, which is an isoform of vitamin E, is a potent fat-soluble phytochemical known in nature. The present study focused on enhancing the production of α-tocopherol in the cell suspension culture through an elicitation approach. Suspension cultures of Nicotiana tabacum were established from the leaf disk-derived callus. The cell suspension cultures were treated with different elicitors (methyl jasmonate, salicylic acid, and yeast extract) at the lag phase of the cell growth cycle. The effects of elicitors on cell cultures were determined in terms of biomass, and α-tocopherol enhancement was determined using high-performance liquid chromatography (HPLC). Different elicitors depending on the concentration exerted different effects on cell growth and α-tocopherol production. Methyl jasmonate treatment showed the significantly highest increase in α-tocopherol on the 6th day of treatment in tobacco suspension cultures. Methyl jasmonate at the concentration of 150 μM enhanced α-tocopherol content to 24-fold over the control. This study clearly shows that the elicitors had the potential to increase the accumulation of α-tocopherol considerably in tobacco cell cultures. The outcomes of this study could be of considerable importance to the nutraceutical and pharmaceutical industries. [ABSTRACT FROM AUTHOR]

Plant tissue culture is the method to produce many pigments, such as anthocyanin, regarding the importance of light in plant growth, especially when using laser rays. The present study aimed to establish the effect of laser rays on seed germination, initiation of callus, and the measurement of the amounts of anthocyanin pigment and protein for the growth of radish (Raphanus sativus L.). Sterilizing seeds were cultured on a surface of MS medium, then seedlings stems were exposed to laser rays for 0, 4, 8, and 12 minutes each alone and planted on MS medium supplemented with 0.5 mg/l of NAA and 1.0 mg/l of BA, the total protein of seedlings was estimated from all kinds of seedlings exposed to the rays. The anthocyanin pigment content was estimated and read Spectroscopically at a wavelength of 528 nm. The results refer to the different rates and periods of radish se ed germination dependent on the exposure time to the laser rays. They also showed the ability of seedlings to initiate callus fr om hypocotyl stems, which was the fastest exposed for 12 minutes after 7 days at a rate of 100 %. The seeds which were previously exposed to diode laser rays at different times recorded a superiority in stimulating anthocyanin pigment content, which amounted to 1195.2 µg/g for stems and 333.8 µg/g for leaves over the rest of the treatments and the control treatment. This proved to be a modern method to develop forms of physical stimulation as a bio elicitor for the growth of radish tissue cells. [ABSTRACT FROM AUTHOR]

Flax (Linum usitatissimum L.) is an important crop plant with pharmaceutical significance. It is described in pharmacopoeias (the United States Pharmacopeia and the European Pharmacopoeia), which confirms that it (especially the seeds) is a valuable medicinal product. Similar to flax seeds, which accumulate bioactive compounds, flax in vitro cultures are also a rich source of flavonoids, phenolics, lignans and neolignans. In the present study, flax suspension cultures after treatment of the non-pathogenic Fusarium oxysporum strain Fo47 were established and analyzed. The study examined the suitability of Fo47 as an elicitor in flax suspension cultures and provided interesting data on the impact of these endophytic fungi on plant metabolism and physiology. Two flax cultivars (Bukoz and Nike) and two compositions of media for flax callus liquid cultures were tested. Biochemical analysis revealed enhanced levels of secondary metabolites (total flavonoid and total phenolic content) and photosynthetically active pigments in the flax callus cultures after treatment with the non-pathogenic fungal strain F. oxysporum Fo47 when compared to control, untreated cultures. In cultures with the selected, optimized conditions, FTIR analysis was performed and revealed changes in the structural properties of cell wall polymers after elicitation of cultures with F. oxysporum Fo47. The plant cell wall polymers were more strongly bound, and the crystallinity index (Icr) of cellulose was higher than in control, untreated samples. However, lignin and pectin levels were lower in the flax callus liquid cultures treated with the non-pathogenic strain of Fusarium when compared to the untreated control. The potential application of the non-pathogenic strain of F. oxysporum for enhancing the synthesis of desired secondary metabolites in plant tissue cultures is discussed. [ABSTRACT FROM AUTHOR]

In recent years, proteomics has become a powerful tool in vitro plant tissue culture research. In Miconia , the most diverse flowering plant genus in the Neotropics, these tools together were unexplored. The group is replete with cases of apomixis (asexual seed formation). Some studies indicate that apomixis may have similarities with somatic embryogenesis (SE). However, there is no investigation into whether explants from apomictic species can affect the accumulation of SE-related proteins in vitro morphogenesis processes, such as callogenesis. This study identifies differentially accumulated proteins related to SE in the callogenesis of three Miconia species with different reproductive systems and whether this can affect their embryogenic capacity. For this, we employed the shotgun comparative proteomics approach in a specific phase of callus proliferation of Miconia australis (obligate apomictic), Miconia hyemalis (facultative apomictic), and Miconia sellowiana (sexual). Comparative proteomic analysis revealed that 190 SE-related proteins were differentially accumulated and distributed among the three experimental comparisons. The calluses of apomictic species showed the highest number of up-accumulated proteins associated with SE. Histological examination of the calluses after 45 days of induction revealed the permanence of cells with meristematic characteristics in the apomictic species, however, in M. sellowiana there was greater cellular vacuolization. These results indicate that calluses of Miconia spp. originated from apomictic species may be better able to acquire embryogenic competence in vitro culture. This study is the first to explore and demonstrate the potential advantages of using proteomics to understand the molecular basis of in vitro morphogenesis in apomictic and sexual species of Miconia , highlighting new perspectives for future research in plant tissue culture. [Display omitted] • Proteomics shed light into the mechanisms underlying in vitro morphogenesis in miconia. • Calluses from apomictic miconia up-accumulated proteins related to somatic embryogenesis. • The cu/zn SOD was up-accumulated in the calluses of obligate apomictic M. australis. • Apomictic species showed a higher prevalence of callus cells with meristematic characteristics. [ABSTRACT FROM AUTHOR]

Nothapodytes nimmoniana (J. Graham) Mabb., commonly known as Narkya, holds immense potential as a medicinal plant, exhibiting a wide array of pharmacological activities. This review provides a comprehensive overview of the methods used for extracting bioactive compounds from different plant parts, along with an exploration of their diverse pharmacological activities. The plant's significance stems from its high content of camptothecin (CPT), a potent anti-cancer alkaloid. We delve into the plant's anti-microbial, antioxidant, anti-tumor, anti-helminthic, anti-malarial, and anti-inflammatory properties. Additionally, we highlight the challenges faced by N. nimmoniana (J. Graham) Mabb., due to overexploitation and its endangered status. To address these concerns, the review underscores the importance of sustainable conservation strategies and the implementation of advanced techniques like plant tissue culture and endophyte production for increased CPT production. This holistic analysis not only sheds light on the plant's multifaceted benefits but also underscores the need for responsible utilization and conservation efforts. [ABSTRACT FROM AUTHOR]

In plant tissue culture, differences in endogenous levels of species-specific plant growth regulators (PGRs) may explain differences in regenerative capacity. In the case of polyamines (PAs), their dynamics and distribution may vary between species, genotypes, tissues, and developmental pathways, such as sexual reproduction and apomixis. In this study, for the first time, we aimed to assess the impact of varying endogenous PAs levels in seeds from distinct reproductive modes in Miconia spp. (Melastomataceae), on their in vitro regenerative capacity. We quantified the free PAs endogenous content in seeds of Miconia australis (obligate apomictic), Miconia hyemalis (facultative apomictic), and Miconia sellowiana (sexual) and evaluated their in vitro regenerative potential in WPM culture medium supplemented with a combination of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP). The morphogenic responses were characterized by light microscopy and scanning electron microscopy and discussed regarding the endogenous PAs profiles found. Seeds of M. sellowiana presented approximately eight times more putrescine than M. australis, which was associated with a higher percentage of regenerated calluses (76.67%) than M. australis (5.56%). On the other hand, spermine levels were significantly higher in M. australis. Spermine is indicated as an inhibitor of auxin-carrying gene expression, which may have contributed to its lower regenerative capacity under the tested conditions. These findings provide important insights into in vitro morphogenesis mechanisms in Miconia and highlight the significance of endogenous PA levels in plant regeneration. These discoveries can potentially optimize future regeneration protocols in Miconia, a plant group still underexplored in this area. [ABSTRACT FROM AUTHOR]

Azadirachta indica or neem is recognized for its unique properties, both against insects and in improving human health. However, proteomics studies of this plant's response to abiotic stresses have yet to be reported. Application of this technique highly depends on efficient and optimized sample preparation, in deriving highly reproducible proteomics data. The present study highlights the comparison of protein extraction method which effectively recovers proteins from recalcitrant phenolic-rich tissues of neem callus. Four protocols based on TCA-acetone or/and phenol extraction methods were compared. The TCA-acetone complemented with PVPP method was the most effective approach and generated the greatest total protein yields, spot number, good resolution, and spot intensities. Subsequently, the differential proteomics performed revealed that proteins associated with oxidoreduction, energy, transcriptional, stress response, respiration, and cell division were differentially regulated in salinity stress treatment. Meanwhile, the expression of proteins classified in the group of energy, photosynthesis, protein synthesis, oxidoreduction, and post-transcriptional regulation was altered in the JA-stressed callus. The accumulation of AZA was also noticeably reduced in salinity- and JA-treated callus, suggesting these tested stress parameters were not effective in improving AZA production in the neem callus. Overall, this analysis provides valuable information on the application of plant tissue culture as an alternative for sustainable production of major bioactive constituents in A. indica. Together, the proteomics results brought forward a possible fundamental representation of the mechanism associated with salinity and JA tolerance in the neem callus. [ABSTRACT FROM AUTHOR]

This study focuses on the endangered neo-endemic Baltic dunes species Linaria loeselii Schweigg. (Plantaginaceae), also known as Linaria odora (M. Bieb.). By utilizing in vitro cultures, we successfully germinated seeds collected in situ. Our method, which involved using media supplemented with 5 µmol/L 6-benzylaminopurine, led to the indirect regeneration of shoots after 60 days of culture in the dark, significantly increasing the number of progeny plants. Additionally, the medium supplemented with 2.85 μmol/L indole-3-acetic acid and 10.2 μmol/L paclobutrazol allowed rooting after 30 days of shoot fragments. This research provides a potential basis for developing Linaria loeselii introduction programs into the environment, thereby contributing to the conservation of this endangered species. [ABSTRACT FROM AUTHOR]

Aim: Potato (Solanum tuberosum L.) is a key crop within the Solanaceae family and ranks as the most significant non-cereal crop globally following major staples such as wheat, rice, and corn. Potatoes can reproduce sexually and asexually via tubers, and plant tissue culture is emerging as an effective method for vegetative propagation, addressing the increasing global demand for agricultural products. Ethylene, a critical plant growth regulator, influences various physiological processes including growth and development. During in vitro culture and due to the wounding of explants, ethylene accumulation can lead to abnormal biological responses, with potato seedlings being susceptible. Thus, investigating the effects of ethylene biosynthesis inhibitors such as pyrazinamide (PZA) and AgNO3 on potato growth in vitro is essential. Material and Methods: In this study, potato seedlings were cultivated in Murashige and Skoog (MS) medium, with concentrations of PZA ranging from 0 to 6 mg L-1 and AgNO3 at 2 mg L-1. After four weeks, the seedlings were harvested and stored at -70°C for later analysis. The growth parameters measured included fresh weight (FW), dry weight (DW), stem and root lengths, leaf area, and leaf and root number. In addition, biochemical parameters, such as photosynthetic pigment levels, total phenol content (TPC), total reactive oxygen species (ROS), and proline concentration were analyzed. Statistical evaluations were conducted using SPSS and PAST software Results: The results showed that the 2 mg L-1 PZA treatment led to the highest FW and DW and increased leaf numbers; however, it was also correlated with a lower number of rooted plants. Conversely, treatments with 6 mg L-1 PZA promoted longer stem growth, whereas control plants exhibited the largest leaf area, and AgNO3-treated plants produced the longest roots. The accumulation of H2O2 in plants treated with ethylene inhibitors was like controls, but total ROS levels soared by 36% in those treated with 6 mg L-1 PZA compared to controls. This suggests a link between reduced ethylene production, oxidative stress mitigation, and enhanced potato growth. Additionally, total ROS was positively correlated with stem length, but negatively correlated with root length. Plants use several strategies to combat the damaging effects of ROS, such as the production of antioxidant compounds such as phenolics. Although PZA did not significantly alter TPC compared to controls, treatment with AgNO3 caused a 61% reduction in TPC. Therefore, PZA did not appear to significantly affect phenolics production in the potato seedlings. Proline, another critical antioxidant in plants, was found to accumulate significantly in the leaves of plants treated with 6 mg L-1 PZA, which was more than 2.3 times higher than that in controls. This accumulation correlated positively with ROS levels at higher PZA concentrations but showed an inverse relationship with photosynthetic pigment levels. The PCA revealed the relationships between the measured parameters and the applied elicitors. The samples were categorized into four distinct groups: 1. Control group: This group primarily exhibited higher FW, DW, and longer roots compared to the treated plants. 2. Low PZA dose group: These plants displayed elevated levels of photosynthetic pigments, TPC, and leaf area. 3. Medium PZA dose group: Correlations were observed with an increased number of roots. 4. 6 mg L-1 PZA and AgNO3 group: These samples contained elevated levels of total ROS and proline. Conclusion: The study concludes that low concentrations of PZA can stimulate growth while inhibiting ethylene production, resulting in fewer growth abnormalities compared to control plants. However, at elevated PZA concentrations, increased ROS levels may lead to oxidative stress, emphasizing the delicate balance in ethylene's role in plant growth and the necessity for further research to optimize conditions for potato cultivation in vitro. The findings contribute to a deeper understanding of how ethylene inhibitors can enhance potato propagation and possibly other crops in controlled agricultural environments. [ABSTRACT FROM AUTHOR]

Background: Lilium (Lilium longiflorum Thunb.) belongs to the family Liliaceae and is a native of Northern Hemisphere (up to South Canada and Siberia). Conventionally, lilium can easily be propagated by sexual and asexual methods of propagation but these prevalent methods are not capable of meeting the increasing demand in domestic and global market. Generally, lilium is propagated through bulbs which has bottlenecks like high cost of planting material, infected from seed borne and other diseases like mosaic, grey mould, etc. So, in vitro multiplication of lilium is utmost important. Lily can be easily regenerated from explants The objective of present study was to identify the best media and the explant among scales and nodal segments belonging to the three different cultivars viz., Elite, Brunello, Cordelia where maximum number of shoots could be regenerated. Methods: The present investigation was carried out in the Tissue Culture Laboratory of the Centre for Research and Application in Plant Tissue Culture. Twenty-five media combinations were used to study their effect on in vitro shoot regeneration from various explants of Lilium longiflorum cultivars. Two type of explants viz. scales and nodal segments were aseptically excised from Elite, Brunello, Cordelia cultivars of Lilum and cultured on Murashige and Skoog (MS) medium containing different concentrations and combinations of 6-benzyl amino purine (BAP), kinetin and Naphthalene Acetic Acid (NAA). Result: In cultivars Elite and Brunello, the maximum number of shoots were observed when nodal segments were used on media combination i.e., LM-8 (MS+BAP 3.0 mg/l+NAA0.1 mg/l). In cultivar Cordelia, scales showed the maximum response on media LM-24 (MS+Kinetin 2.0 mg/l+NAA 0.1 mg/l). [ABSTRACT FROM AUTHOR]

Nanoparticles (NPs) possess strange optical, electrical, and magnetic properties, which arise from their quantum size effects. Nanotechnology, with its immense potential, offers immediate solutions to address these challenges and benefit our society. The pressing concerns of climate change and improving nutritional quality need that we adapt to changing conditions. These distinctive characteristics open up exciting opportunities for the development of innovative sensing techniques that allow for the real-time monitoring of plant responses to nanomaterial exposure. Plant tissue culture is an essential pillar in the field of plant biology, serving as a crucial foundation for a wide range of important applications. This remarkable technique plays a vital role in various areas, such as plant preservation, facilitating large-scale reproduction, enabling genetic modification, fostering the production of bioactive compounds, and enhancing desirable plant characteristics. Through the intricate process of tissue culture, scientists and researchers can manipulate plant cells in a controlled environment, opening up endless possibilities for advancing our understanding of plants and harnessing their potential for benefits. Understanding and optimization of these factors is crucial for improving the efficiency of in vitro propagation. In recent times, the integration of nanoparticles (NPs) has emerged as a successful strategy to combat microbial contaminants in explants, while also showcasing their positive impact on callus initiation, organogenesis, somatic embryogenesis, explants sterilization, and the production of secondary metabolites. This comprehensive review aims to consolidate the significant advancements achieved throughout the integration of nanotechnology into plant tissue culture. It seeks to shed light on the positive attributes associated with the consumption of nanoparticles (NPs) in plant tissue culture, highlighting their enormous potential and benefits. [ABSTRACT FROM AUTHOR]

Paspalum notatum Flüggé is a subtropical grass native from the Americas and one of the main components of the subtropical South American grasslands. Although P. notatum exhibits a high forage productivity and persistence, some limitations are observed in the species, such as forage quality due to high lignin content. The development of efficient tissue culture and plant transformation protocols in cultivars for forage production are necessary to embrace novel genetic improvement through transgenesis and genome editing. Efficient regeneration and transformation systems for many plant species can be hampered by genotype dependency, which can only be determined through extensive laboratory testing and evaluation. In this study, various factors extracted from protocols have been evaluated to establish efficient somatic embryogenesis, plant regeneration, and genetic transformation in apomictic initial variety INIA Sepé. Optimal somatic embryogenesis and regeneration from explant sources, such as mature seeds and meristematic tissue derived from in vitro propagated tillers, ranged between 79 and 88%. Microprojectile bombardment of two plasmids encoding a constitutively regulated reporter gene and nptII selectable marker expression constructs showed an average transformation efficiency of 10.3% from meristematic tissue as an explant source. Furthermore, the constitutive promoters, 35S and ZmUbi, were tested in GFP transient expression assays to evaluate optimal cassettes for transgene(s) expression. Translation of a wheat codon–optimized Cas9 sequence using Cas9:2A:tuGFP fusion protein was also tested in protoplasts. The information generated on promoter activity and Cas9 translation, and the establishment of an efficient transformation system, provided useful tools for future work in genetic engineering and genome editing in P. notatum cv. INIA Sepé. [ABSTRACT FROM AUTHOR]

Juglans mandshurica is an important component of Korean pine mixed and secondary forests in Northeast China and is also a precious timber and economic species. In this study, plant regeneration of J. mandshurica has been established using stems with bud and mature zygotic embryos as explants. The results showed that placing the stems with buds in low-temperature conditions (0 to 4°C) and in darkness for 5 d inhibited the browning of the explants and promoted bud sprouting; Na2S2O3 also inhibited the browning of mature zygotic embryos significantly. The suitable basal media and hormones for bud sprouting and zygotic embryo germination were (1) WPM medium (woody plant medium) containing 2.0 mg·L−1 6-benzylaminopurine (6-BA) and 0.01 mg·L−1 3-indolebutyric acid (IBA) and (2) DKW medium (Driver and Kuniyuki medium) containing 2.5 mg·L−1 6-BA and 0.1 mg·L−1 IBA. The suitable basal medium and hormone for rooting of shoots from buds was 1/2 Murashige and Skoog (MS) medium (the macro-elements were half-strength of MS medium) supplemented with 1.0 mg·L−1 IBA, and the suitable basal medium and hormone for rooting of seedlings derived from zygotic embryo was 1/2 MS medium containing 1.0 mg·L−1 IBA, and DKW medium with 2.5 mg·L−1 6-BA, 0.1 mg·L−1 IBA, 0.2 mg·L−1 gibberellin A3 (GA3), and 0.2 g·L−1 activated carbon (AC). About 95% of the rooted plantlets survived after acclimation and transplantation. These results provided a foundation for the scale propagation to meet the demands of forestry production of the species. [ABSTRACT FROM AUTHOR]

Mass propagation of cassava on several hectares of arable land due to increasing demand for its starch is not feasible due to land availability, pests and disease invasion, and long cultivation period. Plant cell culture technology is a promising solution despite the scarcity of cassava callus culture for starch production applications. Therefore, a systematic mapping study (SMS) was performed to identify the applications of cassava tissue culture and its prospects in starch production and investigate the important parameters for cassava callus culture initiation. The SMS began with formulating research questions (RQs), conducting searches on various databases, collecting and screening related articles, and extracting and mapping the selected articles. A total of 56 of 589 articles in the initial searching phase were chosen to be used as references to answer each RQ. The extracted data indicates that cassava tissue culture was mostly used for micropropagation, while starch production from its tissue culture is still limited. Basal medium and plant growth regulators influence cassava callus culture initiation most. The findings of the SMS offer a better understanding of cassava tissue culture and the prospects of producing cassava starch. [ABSTRACT FROM AUTHOR]

This study examines the effect of ultraviolet-B (UV-B) and ultraviolet-C (UV-C) radiation on contamination rate and shoot proliferation of Tamban pineapple crown explant. The experimental design was nested and completely randomized with a separate control. The first factor was the type of UV light, namely UV-B and UV-C. The second factor was the duration of UV light exposure, namely 10, 20, 30, and 40 min. This study was carried out from March to June 2023 at the Plant Tissue Culture Laboratory, Faculty of Agriculture, Lambung Mangkurat University, South Kalimantan, Indonesia. Observations were made on the contamination percentage, survival percentage, time of first shoot formation, percentage of explants able to regenerate shoots, and number of shoots. The results showed that UV light treatment decreased the contamination rate. Increasing the duration of UV light exposure decreased the contamination rate, delayed the formation of the first shoot, and affected the number of shoots. UV-B light exposure produced a higher number of shoots than UV-C light. These results suggest that UV-B and UV-C radiation have the potential to optimize surface sterilization protocol and promote somaclonal variation. [ABSTRACT FROM AUTHOR]

Plant tissue in vitro culture is increasingly used in agriculture to improve crop production, nutritional quality, and commercial value. In plant virology, the technique is used as sanitation protocol to produce virus‐free plants. Sanitized (S) artichokes show increased vigour compared to their non‐sanitized (NS) counterparts, because viral infections lead to a decline of growth and development.To investigate mechanisms that control the complex traits related to morphology, growth, and yield in S artichokes compared to NS plants, RNAseq analysis and phenotyping by imaging were used. The role of peroxidases (POD) was also investigated to understand their involvement in sanitized plant development.Results showed that virus infection affected regulation of cell cycle, gene expression and signal transduction modulating cellular response to stimulus/stress. Moreover, primary metabolism and photosynthesis were also influenced, contributing to explain the main morphological differences observed between S and NS artichokes. Sanitized artichokes are also characterized by higher POD activity, probably associated with increased plant growth, rather than strengthening of cell walls.Overall, results show that the differences in development of S artichokes may be derived from the in vitro culture stressor, as well as through pathogen elimination, which, in turn, improve qualitative and quantitative artichoke production. [ABSTRACT FROM AUTHOR]

Petiole segments of an economically ornamental plant Oxalis triangularis A.st.-Hil have been used in the present study for rapid, reliable and systematic in vitro propagation. The morphogenic effect of a-Naphthalene acetic acid (NAA) at various concentrations (0.5 - 3.0 mg/L) was studied individually or in combination with 6-benzylaminopurine (BAP). Superior multiplication rates were achieved on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L NAA and 1.0 mg/L BAP. Of the tested combinations, maximum shoot regeneration (100.00 %), mean shoot number (14.60 ± 0.37 shoots per segment), and shoot length (18.11 ± 0.30 mm) were recorded on MS medium supplemented with 0.5 mg/L NAA and 1.0 mg/L BAP after 6 weeks of incubation. Half-strength MS medium augmented with 1.0 mg/L indole-3-butyric acid (IBA) was the optimal medium for in vitro root induction, (24.60 ± 0.37 roots per shoot with 9.10 ± 0.32 mm in length) after 6 weeks of incubation. The regenerated plantlets showed 100 % survival success during acclimatization and exhibited normal growth characteristics and morphology. This optimized protocol is cost-effective and highly efficient for the rapid mass propagation of Oxalis triangularis A.st.-Hil. [ABSTRACT FROM AUTHOR]

Aquatic plants are grown in aquaria for their beauty and to maintain the quality of water. The explants of Lobelia cardinalis L. were obtained from the Aquatic Plant Center Co., Ltd., Thailand. In vitro clonal propagation offers alternative strategy for its rapid clonal multiplication. In this study, multiple and rapid propagations of Lobelia cardinalis L. with tissue culture techniques were aimed. Highly efficient and repeatable in vitro regeneration protocol was established from nodal segments of Lobelia cardinalis L. Various concentrations and combinations of various plant growth regulators (PGRs) were employed to induce multiple shoots, shoot elongation, number of leaves and rooting of shoots to obtain complete plantlets of Lobelia cardinalis L. Among 2 PGRs such as 6-benzylaminopurine (BAP) and a-Naphthalene acetic acid (NAA) tested for multiple shoot induction. Regenerated shoots had a higher number of shoots on MS medium contained cytokinins with auxins. The maximum shoot proliferation (100 %), number of shoots per explant (6.20 ± 0.51 shoots), shoot length (11.67 ± 0.69 mm) and number of leaves per explant (58.10 ± 0.38 leaves per explant) were observed in MS medium supplemented with 1.0 mg/L BAP and 2.0 mg/L NAA. Rooting was achieved within 3 to 4 weeks on all the rooting media, but MS medium augmented with 1.0 mg/L indole-3-butyric acid (IBA) produced the maximum number of strong and healthy roots (11.70 ± 0.27 roots). The regenerated complete plantlets with healthy roots and shoot system were transferred to 3.5 cm diameter black net plastic pots containing rockwool medium and successfully acclimatized with a survival rate of 100 % in the greenhouse and no phenotypic variations were observed among regenerated plants. The success of this protocol offers highly efficient method of the plant multiplication, which would be beneficial for plant tissue culturist, and nursery people where regular supply of plants is a large number required. This protocol can be utilized for commercial scale propagation. [ABSTRACT FROM AUTHOR]

Medicinal plants are invaluable sources of bioactive natural products. To further access plant’s biosynthetic and biotechnological potential, plant tissue culture has become a standard practice. Plant biotechnology might offer selected cell lines with higher metabolic production and allow a more controlled environment for plant development and optimized metabolic flux for the biosynthesis of specific compounds. In this work, we established the in vitro growth of Melia azedarach through micropropagation and evaluated its biosynthetic potential. LC-MS-based metabolic profiling investigation was applied to generate information on natural products composition. A set of computational tools including spectral library matching and in silico prediction of molecular structures and chemical classes were employed to annotate M. azedarach’s plantlets chemical space. Our findings indicate salannin- and nimbolinin-type limonoid production and a series of glycosylated flavonoids and lignan-related phenylpropanoids. This is the first time nimbolinin-related limonoids have been produced in vitro, opening a venue for biotechnological applications of such compounds.Key message: We report the chemical evaluation of natural products diversity of micropropagated Melia azedarach using a combination of metabolomics tools. For the first time, nimbolinin-type limonoids were produced in an in vitro system. [ABSTRACT FROM AUTHOR]

Crocanthemum canadense (L.) Britt. (Cistaceae), Rockrose, is a small perennial herb found in Eastern North America sand barrens. It is classified as critically imperiled in Nova Scotia under the Nova Scotia Endangered Species Act. Nova Scotia C. canadense populations continue to decline; recent counts estimate only 5000–5500 plants remain. To better understand C. canadense conservation, we completed ex-situ in vitro germination and propagation trials using seed collected from native Nova Scotia populations at 14 Wing Canadian Forces Base, Greenwood, Nova Scotia. Concurrently, soil was collected from the same locations to analyze belowground endomycorrhizal symbionts, arbuscular mycorrhizal fungi (AMF). Recent research has documented the presence of AMF within C. canadense roots, however, this is the first study to successfully identify AMF species present in-situ using single spore rDNA barcoding. C. canadense successfully germinated in vitro with high germination rates (average 86.6%) for one population (SB000147) maintained at room temperature with a 16-hour photoperiod. Successful vigor continued to be monitored with regular multiplication and transfers. Eight AMF spores were identified, all members of the genus Rhizophagus. Seven spores identified as Rhizophagus clarus and one as Rhizophagus irregularis. R. clarus are commonly found in many semi-arid environments now including sand barrens. This research will aid in the conservation and restoration of this critically imperiled species by further understanding beneficial soil fungi, an understudied component of the declining sand barrens habitat. [ABSTRACT FROM AUTHOR]

Solanum nigrum (Solanaceae family) is widely consumed as a fruit or local leafy vegetable after boiling; it also serves as a medicinal plant. Although Agrobacterium-mediated genetic transformation has been established in S. nigrum, the transformation period is long. Specifically, induction of roots takes approximately five weeks for tetraploid and hexaploid S. nigrum, and 7 weeks for diploid Solanum americanum. In this study, we developed an improved rooting-induced method that requires only about 1 week and avoids the use of tissue culture. After generating the transgenic shoots, they were directly transplanted into the soil to facilitate root formation. Remarkably, 100% of the transgenic shoots developed roots within 6 days. Our improved method is time-saving (saving more than 1 month) and simpler to operate. The improved rooting-induced step can be applied to induce roots in various plants using tissue culture, exemplified by the carnation (Dianthus caryophyllus L.). Furthermore, we applied the improved method to generate S. americanum plants expressing AcMYB110 from kiwifruit (Actinidia chinensis spp.). This method will contribute to speeding up gene functional analysis and trait improvement in S. nigrum and might have potential in fast plant molecular breeding processes in crops and rapid rooting induction in tissue culture. [ABSTRACT FROM AUTHOR]

[Objective] To compare the effectiveness of 8 brands of powder MS medium, so as to provide a reference basis for selecting medium for plants tissue culture. [Method Using the leaves, petioles, roots of E. purpurea and stem segments of A. roxburghii as explants, the standard MS and the national authorized invention patent medium without explosive compounds were used as controls, the effectiveness of 8 brands of powder MS medium on callus induction, adventitious bud regeneration, and rooting of E. purpurea and A. roxburghii was compared. [Result] The three brands of powder MS medium codes M5, M6 and M8 were more effective in callus induction of E. purpurea, and no significant difference compared with the two controls (CK1 and CK2), whereas the other five brands were significantly worse than the control. The results of the experiment on inducing adventitious bud of E. purpurea showed that five brands medium codes M1, M2, M3, M4, and M7, failed to induce adventitious buds either by using leaves or petioles as explants in E. purpurea, while the M5 medium not only successfully induced but also obtained more adventitious buds than the control. On the other hand, the comparative experiment of A. roxburghii tissue culture showed that all 10 mediums could successfully induce to obtain adventitious buds, and the induction rate of adventitious buds in M4 was higher than the control, reaching 83.5%. The results of inducing adventitious root in E. purpurea showed that only the medium codes M5 and M8 could successfully induce adventitious roots from adventitious buds as well as the two controls, but the number and length of adventitious roots induced by M5 medium were lower than the two controls. In the experiment of inducing lateral bud rooting of A. roxburghii, all 8 brands medium could successfully induce adventitial roots, and the number and length of adventitious roots induced by M2 were significantly higher than the control. [Conclusion] The effects of 8 brands of powder MS medium were significant different in the tissue culture of E. purpurea and A. roxburghii, and except for the medium codes M5, M6, and M8, other brands of powder MS medium are not suitable for tissue culture of E. purpurea. However, except for M4, all 7 other brands of powder MS medium are suitable for A. roxburghi. [ABSTRACT FROM AUTHOR]

Background: Microbial growth during plant tissue culture is a common problem that causes significant losses in the plant micro-propagation system. Most of these endophytic microbes have the ability to propagate through horizontal and vertical transmission. On the one hand, these microbes provide a rich source of several beneficial metabolites. Results: The present study reports on the isolation of fungal species from different in vitro medicinal plants (i.e., Breynia disticha major, Breynia disticha, Duranta plumieri, Thymus vulgaris, Salvia officinalis, Rosmarinus officinalis, and Ocimum basilicum l) cultures. These species were tested for their indole acetic acid (IAA) production capability. The most effective species for IAA production was that isolated from Thymus vulgaris plant (11.16 µg/mL) followed by that isolated from sweet basil plant (8.78 µg/mL). On screening for maximum IAA productivity, medium, "MOS + tryptophan" was chosen that gave 18.02 μg/mL. The macroscopic, microscopic examination and the 18S rRNA sequence analysis indicated that the isolate that given code T4 was identified as Neopestalotiopsis aotearoa (T4). The production of IAA by N. aotearoa was statistically modeled using the Box-Behnken design and optimized for maximum level, reaching 63.13 µg/mL. Also, IAA extract was administered to sweet basil seeds in vitro to determine its effect on plant growth traits. All concentrations of IAA extract boosted germination parameters as compared to controls, and 100 ppm of IAA extract exhibited a significant growth promotion effect for all seed germination measurements. Conclusions: The IAA produced from N. aotearoa (T4) demonstrated an essential role in the enhancement of sweet basil (Ocimum basilicum) growth, suggesting that it can be employed to promote the plant development while lowering the deleterious effect of using synthetic compounds in the environment. [ABSTRACT FROM AUTHOR]

Notwithstanding the fact that melatonin (MT) and titanium nanoparticles (Ti NPs) alone have been widely used recently to modulate cadmium (Cd) stress in plants, there is a gap in the comparative impacts of these materials on lowering Cd toxicity in sage plants. The objective of this study was to determine how foliar application of MT and Ti NPs affected the growth, Cd accumulation, photosynthesis, water content, lipid peroxidation, and essential oil (EO) quality and quantity of sage plants in Cd-contaminated soils. A factorial experiment was conducted using MT at 100 and 200 μM and Ti NPs at 50 and 100 mg L−1 topically, together with Cd toxicity at 10 and 20 mg Cd kg−1 soil. The results showed that Cd toxicity decreased plant growth and enhanced lipid peroxidation. The Cd stress at 20 mg kg−1 soil resulted in increases in Cd root (693%), Cd shoot (429%), electrolyte leakage (EL, 29%), malondialdehyde (MDA, 72%), shoot weight (31%), root weight (27%), chlorophyll (Chl) a + b (26%), relative water content (RWC, 23%), and EO yield (30%). The application of MT and Ti NPs controlled drought stress by reducing MDA and EL, enhancing plant weight, Chl, RWC, and EO production, and minimizing Cd accumulation in plant tissues. The predominant compounds in the EO were α-thujone, 1,8-cineole, β-thujone, camphor, and α-humulene. MT and Ti NPs caused α-thujone to rise, whereas Cd stress caused it to fall. Based on heat map analysis, MDA was the trait that was most sensitive to treatments. In summary, the research emphasizes the possibility of MT and Ti NPs, particularly MT at 200 μM, to mitigate Cd toxicity in sage plants and enhance their biochemical reactions. [ABSTRACT FROM AUTHOR]

A protocol for optimization of callus culture establishment from leaf explants of Portulaca oleracea L. was developed. The effect of ethyl methanesulphonate (EMS) on callus induction and synthesis of fatty acids was evaluated. Callus culture was initiated from leaf explants on Murashige and Skoog’s medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D), a combination of 2,4-D + 6-benzyl adenine (BA) or 2,4-D + kinetin. The maximum callus biomass was obtained at 2.5 µM BA + 2.5 µM 2,4-D (20.22 g leaf explant-1 fresh weight). The leaves treated with EMS (0.1 to 0.4% h-1) differentiated callus on this optimized medium combination. The gas chromatography-mass spectrometry analysis of fatty acids indicated that the leaves had a high linolenic acid content (17.72%). The callus cultures synthesized heptadecanoic, oleic, and tricosanoic acids, which were otherwise absent in the leaves. Eicosanoic and docosanoic acids in callus cultures were 2.78 and 3.18 times higher than their content in the leaves, respectively. The diversity of fatty acids in treated callus cultures decreased with increased EMS concentration, but the content of a few individual fatty acids was enhanced. Callus at the dose of 0.1% EMS synthesized linoleic acid, which was two times higher than in the untreated callus, while at 0.2% EMS stearic acid was synthesized, which was absent in the untreated callus. It was concluded that the callus of P. oleracea accumulated different fatty acids, and EMS treatment enhanced the content of a few fatty acids in the cultures. [ABSTRACT FROM AUTHOR]

Since prehistoric times, medicinal and aromatic plants (MAPs) have been employed for various therapeutic purposes due to their varied array of pharmaceutically relevant bioactive compounds, i.e. secondary metabolites. However, when secondary metabolites are isolated directly from MAPs, there is occasionally very poor yield and limited synthesis of secondary metabolites from particular tissues and certain developmental stages. Moreover, many MAPs species are in danger of extinction, especially those used in pharmaceuticals, as their natural populations are under pressure from overharvesting due to the excess demand for plant‐based herbal remedies. The extensive use of these metabolites in a number of industrial and pharmaceutical industries has prompted a call for more research into increasing the output via optimization of large‐scale production using plant tissue culture techniques. The potential of plant cells as sources of secondary metabolites can be exploited through a combination of product recovery technology research, targeted metabolite production, and in vitro culture establishment. The plant tissue culture approach provides low‐cost, sustainable, continuous, and viable secondary metabolite production that is not affected by geographic or climatic factors. This study covers recent advancements in the induction of medicinally relevant metabolites, as well as the conservation and propagation of plants by advanced tissue culture technologies. [ABSTRACT FROM AUTHOR]

Unable to move on their own, plants have acquired the ability to produce a wide variety of low molecular weight compounds to survive against various stresses. It is estimated that there are as many as one million different kinds. Plants also have the ability to accumulate high levels of proteins. Although plant-based bioproduction has traditionally relied on classical tissue culture methods, the attraction of bioproduction by plants is increasing with the development of omics and bioinformatics and other various technologies, as well as synthetic biology. This review describes the current status and prospects of these plant-based bioproduction from five advanced research topics, (i) de novo production of plant-derived high value terpenoids in engineered yeast, (ii) biotransformation of plant-based materials, (iii) genome editing technology for plant-based bioproduction, (iv) environmental effect of metabolite production in plant factory, and (v) molecular pharming. [ABSTRACT FROM AUTHOR]

Tissue culture techniques were used to produce large amounts of bioactive compounds with medicinal potential, overcoming space and time constraints for cancer prevention. Rice callus suspension cultures (RCSC) and seed extracts prepared from aromatic rice varieties were used to evaluate the cytotoxic impact on human colon and lung cancer cell lines, as well as a normal control cell line, using Taxol as a positive control. RCSC and seed extracts from two Indian aromatic rice varieties were applied at different concentrations to treat the cancer cell lines and normal lung fibroblasts over varying time intervals. Apoptosis was assessed in 1:5 dilutions of the A549 and HT-29 cell lines treated with RCSC for 72 h, using propidium iodide staining and flow cytometry. RCSC showed a more potent cytotoxic effect than seed extracts with minimal effect on the normal cell line, in contrast to Taxol. Confocal microscopy and flow cytometry further confirmed the apoptotic effect of RCSC. Gas chromatography-mass spectrometry-based metabolic profiling identified metabolites involved in cytotoxicity and highlighted altered pathways. RCSC is proposed as an alternative source for the development of novel anticancer drugs with reduced side effects. [ABSTRACT FROM AUTHOR]

Turmeric has attracted a significant amount of interest in recent years due to its strong antimicrobial properties. The tissue culture of turmeric is preferred to obtain disease-free, highest number of plantlets with good uniform chemistry. However, there is a need to increase the speed of the whole process to meet the growing demand for planting materials and to save time and resources. Iron oxide nanoparticles (Fe3O4 NPs) showed positive effects on callus initiation time, proliferation rate, percent root response, shoot length, percent rooting, and number of roots per explant. Highest callus induction, i.e., 80%, was recorded in cultures that were grown in the presence of 15 mg/L of Fe3O4 NPs. Callus initiated earlier in culture tubes that received green synthesized iron nanoparticles in a concentration between 10–15 mg/L. Biofabricated nanoparticles were characterized for their size, physiochemical, and optical properties through UV–Vis spectroscopy, FTIR, XRD, and SEM. Curcuminoids profiling was performed by implementing LC-Ms that revealed increased quantities in plantlets grown in nano-supplemented media when compared to the control. [ABSTRACT FROM AUTHOR]

The significant issue of managing in-vitro culture browning in plant tissue culture procedures is explored in depth in this review. The complex issue of browning, which includes oxidative, enzymatic, and non-enzymatic browning, poses obstacles to the effectiveness of in vitro culture and has an impact on the quality of secondary metabolites as well as plant viability. Researchers and practitioners encounter this issue in their pursuit of successful plant propagation and valuable secondary metabolite production. The review explores a range of control measures, including antioxidants like ascorbic acid, polyvinylpyrrolidone (PVP), and potassium citrate, alterations to the culture medium, and the selection of suitable culture vessels. These techniques provide a comprehensive method for controlling browning and getting good results in plant tissue culture. The knowledge gathered from this research advances the industry and promotes sustainability since plant tissue culture is still essential for creating disease-free plants, quickly proliferating uncommon genotypes, and synthesizing useful plant-derived compounds. Whether enhancing crop traits or cultivating plants with economically significant products, effective control of culture browning remains a key factor in achieving these goals. In conclusion, this review advances our understanding of the multifaceted challenge of culture browning in plant tissue culture and provides actionable solutions to address this issue. By implementing these measures, researchers and practitioners can enhance the outcomes of plant tissue culture, maximize the yield of valuable products, and contribute to the growth and sustainability of agriculture and industry. [ABSTRACT FROM AUTHOR]