Long non-coding RNAs (lncRNAs) are emerging as important regulators of plant gene expression, affecting developmental processes, stress tolerance, and organ growth. Their involvement in the determination of seed size is yet to be explored in litchi (Litchi chinensis). An in-depth lncRNA profiling of developing seeds of large- and small-seeded litchi cultivars was conducted and 1,014 lncRNAs, of which 88 were found to be differentially expressed, was discovered. A few lncRNAs, such as XLOC_030992, XLOC_021376, and XLOC_025154, exhibited remarkable regulatory capacity through co-expression or anti-correlation with genes of hormone signalling, intracellular transport, proteostasis, lipid metabolism, and cell cycle regulation. Interestingly, XLOC_030992 appeared to be a hub trans-acting regulator of more than 180 target genes, some of which are implicated in ABA signaling, mitotic progression, and seed viability. Others, including XLOC_009460 and XLOC_006988, showed stage- and genotype-specific expression, indicating dynamic functions in early embryogenesis. This study’s results suggested a regulatory model in which lncRNAs regulate seed development by cis- and trans-regulating important genes. This research provided the first evidence for the role of lncRNAs in litchi seed size determination and provided a basis for functional studies to improve seed quality in fruit crops.
Nodal stem explants of Epidendrum radicans Pav. ex Lindl. showed two distinct in vitro regeneration routes under different plant growth regulators. This study compared these routes, characterized the 2,4-D-induced structures histologically, and evaluated their use in a two-stage micropropagation strategy. Stable aseptic cultures were obtained most effectively with 0.1
Meconopsis aculeata Royle holds ecological, medicinal, and ornamental value yet faces conservation challenges due to limited natural populations and insufficient propagation techniques. Therefore, the present investigation evaluates the seed germination, in vitro propagation, acclimatization, and biochemical characteristics of in vitro grown plants of M. aculeata. Optimal seed germination was accomplished at 25°C on MS basal medium and with a maximum germination percentage (approximately 70
Pineapple (Ananas comosus L. Merr., cultivar MD2) is a key tropical crop but highly sensitive to salinity and drought, making temporary immersion bioreactors (TIBs) valuable for micropropagation and stress studies. Apical buds were cultured in vitro, transferred to TIBs with paclobutrazol, and subjected to nine treatments (control, NaCl, mannitol at 50.0 to 200.0 mM) for 30 d. Growth and biochemical traits were measured, data normalized (0 to 1), and analyzed by hierarchical cluster analysis (HCA) using average linkage and squared Euclidean distance. The dendrogram revealed clear treatment similarities: controls remained distinct until higher rescaled distances, low concentrations clustered together, and high concentrations formed separate groups, confirming dose-dependent responses. NaCl and mannitol did not cluster together initially, indicating distinct physiological profiles, with NaCl imposing additional ionic toxicity beyond osmotic effects. These findings complement earlier evaluations showing significant reductions in shoot multiplication and biomass above 50.0 mM, with critical concentrations for 50
Artemisia vulgaris L. has attracted considerable interest for its pharmaceutical, agricultural, and cosmetic applications due to its rich bioactive metabolites. However, low metabolite yields and challenges in large-scale cultivation constrain its commercial potential. Therefore, this study aimed to optimize a callus regeneration system, develop a comprehensive metabolite profile of the callus culture, and assess the effects of elicitor treatments on metabolite biosynthesis pathways to identify optimal concentrations for enhanced metabolite production. In vitro callus cultures were initiated on Murashige and Skoog (MS) medium with varying levels of benzylaminopurine (BAP), α-naphthaleneacetic acid (NAA), and 2,4-dichlorophenoxyacetic acid (2,4-D) phytohormones. Callus cultures were evaluated for colour, texture, moisture content, and biomass, and metabolite profiling was conducted using nuclear magnetic resonance (NMR) platform. Elicitor treatments of the callus cultures with different concentrations of methyl jasmonate (MJ) and salicylic acid (SA) were analysed via NMR-based metabolomics. MS medium containing 2.0 mg L⁻1 BAP and 0.2 mg L⁻1 NAA was optimal for callus culture establishment and maintenance. A total of eighteen compounds were identified, including dihydroartemisinic acid, caffeic acid, and gallic acid. MJ and SA functioned as stress elicitors, modulating the tricarboxylic acid cycle, phenylpropanoid, and artemisinin biosynthesis pathways, although they had limited direct effects on artemisinin concentrations. Overall, this study optimized callus culture production, generated the first detailed metabolite profile, and highlighted the potential of elicitation strategies to enhance bioactive metabolite yields, providing a foundation for future metabolic engineering and industrial applications.
An efficient micropropagation method based on axillary shoot bud proliferation was developed for Barleria prionitis, an important Indian woody medicinal plant. Highest (100
Alocasia cuprea, a rare ornamental aroid from Borneo, has high commercial value but is constrained by slow natural propagation. This study presents an integrated in vitro morphogenesis framework, examining the combined effects of explant type, culture system, plant growth regulators, and incubation conditions on callus initiation, proliferation, and regeneration. Morphogenic outcomes were strongly determined by the interaction of medium composition, culture system, and explant type (p < 0.001). During initiation, corm explants cultured on Murashige and Skoog (MS) medium with 1.0 mg L⁻1 thidiazuron (TDZ) and 0.5 mg L⁻1 6-benzylaminopurine (BAP) under a bilayer system achieved the highest callus formation (74.5
Apospory, the formation of diploid gametophytes directly from sporophytic tissue without meiosis, represents a rare deviation in the diplohaplontic life cycle of bryophytes with important ecological and evolutionary implications. In this study, a protocol for apospory induction in the moss Physcomitrium eurystomum, a rare and endangered species of allopolyploid origin was established. Differentiation of diploid gametophytic protonemata arose from medulla tissue of sporophytic setae, and induction was strongly dependent on the seta developmental stage. Injury to sporophytic tissue, whether by cutting or mechanical damage, further stimulated differentiation, underscoring the role of wounding as a trigger of apospory. The resulting diploid gametophytes exhibited larger phylloids and significantly larger cell sizes compared to haploids, while maintaining normal pigment composition and physiological function. Cytogenetic analyses confirmed their diploid status, validating successful apospory induction. These results provide a framework for generating clonal diploid lineages in mosses and highlight apospory as an important process for understanding genome–environment interactions, stress tolerance, and the evolutionary flexibility of alternation of generations in bryophytes.
Eclipta alba (L.) Hassk. is a medicinally important plant, known for its pharmacologically valuable bioactive secondary metabolites, particularly wedelolactone, flavonoids, and phenolic acids. The present study investigated the effect of biosynthesized copper oxide (CuO) and zinc oxide (ZnO) nanoparticles (NPs) on E. alba in vitro cultures supplemented with different concentrations (0.0, 2.0, 4.0, 6.0, 8.0, 10.0, and 12.0 mg L⁻1). Both nanoparticle types exhibited concentration-dependent biphasic responses across all evaluated parameters with optimal stimulatory effects at 8 mg L⁻1. At this concentration, CuO nanoparticles enhanced fresh weight (4.1-fold), dry weight (5.9-fold), total phenolic content (TPC; 7.4-fold), total flavonoid content (TFC; 2.3-fold), total antioxidant capacity (TAC; 4.5-fold), total reducing power (TRP; 6.1-fold), and total protein content (2.8-fold), while ZnO NPs produced comparable enhancements of fresh weight (3.9-fold), dry weight (5.3-fold), TPC (7.7-fold) and TFC (2.6-fold), TAC (4.6-fold), TRP (6.4-fold), and total protein content (2.1-fold) relative to control. Maximum DPPH radical scavenging activity (96.85
Holmskioldia sanguinea is valued for its ornamental appeal and traditional medicinal uses for treating rheumatism, dysentery, headaches, hypertension, ulcers, and gynecological disorders, but propagation challenges limit wider cultivation and conservation efforts. To overcome the limitations associated with conventional propagation methods, an effective in vitro regeneration protocol was established using nodal explants. Following surface sterilization, explants were cultured on half strength Murashige and Skoog (MS) medium supplemented with varying concentrations of plant growth regulators for shoot bud initiation and multiplication. The best response was observed with 1.0 mg L−1 BA (Benzyladenine) particularly when combined 0.5 mg L−1 2,4-D (2,4- Dichlorophenoxyacetic acid) Shoot formation was evident within 2 to 3 wk, and rooting was successfully achieved in two wk after transferring the explant with initiated shoots on half strength MS medium supplemented with 1.0 mg L−1 IBA (Indolebutyric acid). Acclimatization involved a two-phase hardening process, beginning with a gradual reduction in external nutrients and followed by transfer to sterile peat moss + sand (2:1, v/v) potting mix, resulting in an impressive 98
Asarum sieboldii Miq. is an important medicinal plant widely used in traditional East Asian medicine for its analgesic, anti-inflammatory, and antimicrobial properties. However, its large-scale utilization is constrained by slow natural propagation and excessive harvesting from wild populations. To address these challenges, this study reported the first efficient and reproducible in vitro propagation protocol for A. sieboldii. Apical shoot explants were cultured to evaluate the effects of basal media, plant growth regulators, and acclimatization conditions on regeneration efficiency. Full-strength Murashige and Skoog (MS) medium was identified as the most suitable basal medium for culture establishment and shoot development. Cytokinin optimization demonstrated that MS medium supplemented with 1.5 mg L−1 6-benzylaminopurine (BAP) produced optimal shoot proliferation, yielding 5.8 ± 0.83 shoots and 12.25 ± 1.25 leaves per explant after six wk. Root induction was significantly influenced by auxin type and combination, with the highest rooting efficiency achieved on MS medium containing 0.75 mg L−1 naphthaleneacetic acid (NAA) and 1.0 mg L−1 indole-3-butyric acid (IBA). FT-NIR spectral and multivariate chemometric analyses revealed high biochemical similarity between mother and in vitro regenerated plants, with metabolic variation driven primarily by organ type rather than propagation method. Flow cytometric analysis confirmed the maintenance of genome size and ploidy levels, indicating genetic stability of regenerated plants. Morphological observations further demonstrated that in vitro regenerated plants exhibited phenotypic characteristics comparable to mother plants with no visible abnormalities. Acclimatization was highly successful, achieving a 98
Marigolds (Tagetes spp.) are widely cultivated for ornamental, medicinal, and industrial purposes, yet efficient regeneration protocols, particularly for French marigold (Tagetes patula L.), remain limited, hindering genetic improvement efforts. This study reports the development of an optimized regeneration system for African (Tagetes erecta L.) and French marigolds through indirect organogenesis using cotyledon explants. Nine cultivars were evaluated for callus induction and shoot regeneration under varying auxin-to-cytokinin ratios using naphthaleneacetic acid (NAA) and benzylaminopurine (BAP). Genotype-specific responses were observed in both callus induction and shoot regeneration. Callus induction was most effective in ‘Bonanza Flame’ and ‘Marvel II Yellow’ at a 1:1 NAA:BAP ratio (1.0 mg L−1 each). Shoot regeneration reached up to 80
The expansion of genome-editing tools for bread wheat (Triticum aestivum L.) is essential to accelerate trait improvement while addressing intellectual property constraints associated with widely used CRISPR systems. MAD7 (ErCas12a), a royalty-accessible CRISPR nuclease, represents a potential alternative to CRISPR-Cas9; however, its performance in complex polyploid crops remains insufficiently characterized. In this study, the in planta genome-editing efficiency of MAD7 in hexaploid wheat was evaluated using Agrobacterium-mediated transformation. Conserved coding regions of the TaLCYε (LYCOPENE EPSILON CYCLASE) gene, a key regulator of carotenoid flux, were targeted across all three wheat subgenomes (A, B, and D). MAD7-mediated editing showed strong dependence on protospacer adjacent motif (PAM) composition, with detectable activity only at a T-rich PAM (TTTG), resulting in targeted mutations in 26
Hairy root cultures of Phyllanthus acuminatus Vahl represent a sustainable system for producing bioactive metabolites such as glycosides, lignans, and phenolics. This study evaluated the effect of methyl jasmonate (50.0 µM) and salicylic acid (50.0 and 200.0 µM) on biomass accumulation and culture medium conductivity in hairy roots of P. acuminatus. The highest dry weight (172.07 mg) and dry/fresh ratio (11.5
The rising requirement for plant-derived bioactive molecules has driven interest in sustainable production. In the present investigation, biosynthetic enhancement of pharmacologically relevant secondary metabolites was achieved using Aerva lanata, a therapeutic species with established ethnopharmacological relevance but limited utilization. Comparative evaluation of five Agrobacterium rhizogenes strains identified R1000 as the most effective, producing a transformation frequency of 86.2 ± 0.2
Bulbous ornamental plants are of great importance in plant biotechnology and conservation biology due to their ecological functions and high economic value. This study was carried out to evaluate the hormone, culture medium, and light factors affecting bulblet formation in Lilium akkusianum, a locally endemic and endangered plant, under in vitro culture conditions. Leaf and root explants were examined in two culture media (MS and B5), different 2,4-D (0.1, 1.0, and 2.0 mg L⁻1) and kinetin (0.1, 1.0 mg L⁻1) doses, and two different climate conditions (25 ± 2 °C temperature and 15 d of continuous darkness, 25 ± 2 °C temperature, 16/8 h light/dark, and 3.000 lx light intensity). Results showed that MS medium provided a higher regeneration capacity compared to B5 medium. Leaf explants formed more bulblets than root explants. The highest bulblet formation was obtained from leaf explants with the combination of MS containing 2.0 mg L−1 2,4-D and 1.0 mg L−1 kinetin (8.33 pieces) under dark conditions. In root explants, MS with 1.0 mg L−1 2,4-D and 0.1 mg L−1 kinetin medium and dark conditions (13.3 pieces) were the prominent combination. These results suggested that an auxin-cytokinin balance plays a critical role in organogenesis and that light conditions significantly affect the embryogenic response. This study provided a scientific basis for the development of an effective micropropagation protocol for this local endemic species and contributed to the conservation of the species and to the increase of its production potential as an ornamental plant.
Banana (Musa spp.) is a globally important staple and cash crop where an efficient anther culture protocol has been developed to enable the production of doubled haploid (DH) lines for use in breeding programs and genomic research. This study compared the effectiveness of different culture media Murashige and Skoog (MS), N6, and Nitsch in inducing embryogenic callus formation from anthers of five different banana genotypes across three genomic groups (BB, AA, AAA). Among the media tested, MS medium proved to be most effective for embryogenic callus induction. Among the five genotypes tested, Bhimkol (BB) produced embryogenic callus on MS medium supplemented with 4.0 mg L−1 2.4-Dichlorophenoxyacetic acid (2,4-D) and 1.0 mg L−1 each of Indole-3-acetic acid (IAA), and α-Naphthaleneacetic acid (NAA). The callus were sub-cultured three times at regular intervals onto MS medium fortified with 2.0 mg L−1 IAA and 0.5 mg L−1 6-Benzyl amino purine (BAP). The resultant shoots were then transferred to MS medium supplemented with 2.0 mg L−1 BAP and 0.5 mg L−1 IAA to enhance shoot multiplication and elongation. Rooting was successfully achieved by culturing the single shoots on MS medium containing 1.0 mg L−1 each of Indole-3-butyric acid (IBA) and NAA. The rooted plantlets were acclimatized successfully, with 60
Efficient and reproducible regeneration procedures are important for the establishment of reliable in vitro systems in medicinal plants. Here, a central composite design (CCD) consisting of 30 treatments was employed for modelling and optimisation of combined influences of kinetin (KI), 6-benzylaminopurine (BAP), indole-3-acetic acid (IAA) and maltose treatments on direct shoot regeneration from immature leaf explants of Enicostemma littorale Blume. Two responses were evaluated, which were the shoot regeneration frequency (Y1
To establish and improve the tissue culture system of 'Autumn–Winter Yellow Raspberry', this study optimized the combinations of plant growth regulators for shoot proliferation and rooting, compared transplanting performance of plantlets under different culture durations, and preliminarily established an adventitious bud regeneration system. The optimal proliferation medium was Murashige and Skoog (MS) with 0.6 mg L⁻1 6-benzylaminopurine (6-BA), 0.3 mg L⁻1 indole-3-butyric acid (IBA) and 30.0 g L⁻1 sucrose, pH = 5.9. The optimal rooting medium was modified 1/2MS with 0.8 mg L⁻1 IBA and 30.0 g L⁻1 sucrose, pH = 5.9, and the best transplant survival rate was obtained after 30 d of culture in this rooting medium. When leaf explants were cultured on MS with 1.0 mg L⁻1 thidiazuron (TDZ) and 30.0 g L⁻1 sucrose, pH = 5.9 with the adaxial side down in darkness for 21 d, and then transferred to light, the induction rate exceeded 90
Japanese plum varieties demonstrate varying levels of resistance to black knot disease, which is caused by the fungal pathogen Apiosporina morbosa (Schwein van Arx.). The fungus produces unsightly black cankers, resulting in reduced tree vigor and dieback. In this study, we employed an untargeted LC–MS/MS metabolomics approach to analyze the metabolome profiles of stems from two resistant varieties (‘Underwood’ and ‘Redcoat’) and two susceptible varieties (‘Vampire’ and ‘Shiro’) at various stages of infection. A total of 468 compounds were identified, with notable differences in the quantities of catechin and 3,4-dihydroxybenzaldehyde between resistant and susceptible varieties. The highest levels of these compounds were detected in June, coinciding with the onset of visible disease symptoms. These observations were further validated with HPLC and in vitro antifungal assay, strengthening the association between these metabolites and black knot resistance. Catechin and 3,4-dihydroxybenzaldehyde are both recognized for their antimicrobial effects, suggesting that their increased accumulation may contribute to disease resistance by suppressing fungal infection. Overall, our findings indicate that these compounds play a pivotal role in black knot resistance in Japanese plums and present promising targets for improving disease resistance through selective breeding.
