Search Results (164)

The chemical profile of coffee depends on numerous factors, the complexity of which makes it difficult to clearly assess their influence. The aim of this study was to comprehensively evaluate the impact of selected coffee brewing methods (Espresso, Simple Infusion, French Press, V60), taking into account the coffee species (Arabica, Robusta, Blends), the degree of roasting (light, medium, dark) and the geographical origin (single-origin and multi-origin) on the chemical composition of the brew. Eighteen different types of coffee, which differ in the aforementioned characteristics, were analyzed. The caffeine content (using high-performance liquid chromatography), the total phenolic content (TPC; using a spectrophotometric method), and selected minerals (calcium, iron, potassium, magnesium, sodium, phosphorus, zinc; using Inductively Coupled Plasma–Optical Emission Spectrometry) were analyzed. The analysis showed that both the brewing method and the species had a significant influence on the chemical profile of the resulting brews, while the degree of roasting and the origin showed no significant influence. The Espresso method showed the highest caffeine, TPC, potassium, magnesium, and phosphorus content, the V60 method—calcium, iron, and sodium, and the French Press and Simple Infusion methods showed intermediate values. Robusta coffee contained more caffeine and TPC, Arabica contained more magnesium, and Blend showed medium values for both species. The results obtained may have practical implications for both consumers and the coffee industry, supporting informed decision-making and the refinement of brewing methods. Full article

Quercus suber L. (Q. suber) is an ecologically and industrially valuable species, yet faces challenges in propagation in China. This study optimized somatic embryogenesis (SE) protocols using two-year-old Q. suber leaves, focusing on petioles and leaf veins as the most responsive explants, with May as the optimal sampling time. The MSSH medium (a combination of Murashige and Skoog Medium (MS) major elements and Schenk and Hildebrandt Medium (SH) minor elements and vitamins) under darkness maximized transdifferentiation. Additionally, the highest callus induction was achieved with 0.50 mg/L 6-benzyladenine (6-BA) and 1.00 mg/L 1-Naphthaleneacetic acid (NAA). Liquid culture with 1.00 g inoculum and 0.50 mg/L 6-BA + 0.20 mg/L NAA achieved the best proliferation. Redifferentiation peaked at 0.15 mg/L NAA + 0.20 mg/L 6-BA. Transcriptome profiling identified 4534 differentially expressed genes (DEGs) between embryogenic callus (E1) and global embryos (E2), with key pathways linked to cell wall remodeling, stress responses, and photosynthesis. Key regulators identified during the early stage of callus redifferentiation include cytokinin oxidase 3 (CKX3), gibberellin-responsive protein (GH3.6), and pectin lyase 5 (PL5), among others. This study provides insights into efficient SE of Q. suber and the genes underlying early callus redifferentiation, laying the groundwork for future research. Full article

Ganoderic acid (GA) is a key bioactive component with pharmacological properties that is found in Ganoderma lingzhi, a renowned medicinal mushroom. Currently, the regulatory mechanisms underlying GA biosynthesis in G. lingzhi remain to be further elucidated. In this study, blue light induction was found to significantly enhance the GA content in G. lingzhi. To explore the regulatory mechanism of GA biosynthesis in response to blue light, the blue light receptor WC-2 was identified, and its regulatory role was characterized. The deletion of wc-2 resulted in a significant reduction in both GA content and the accumulation of intermediates compared to the wild-type control strain, largely due to the strong downregulation of key GA biosynthetic genes. Additionally, decreased asexual spore production and reduced expression of sporulation-specific genes were observed with the deletion of wc-2. The overexpression of wc-2 led to greatly enhanced GA accumulation. Under blue light induction, the maximum contents of GA-Mk, GA-T, GA-S, and GA-Me were 2.27-, 2.51-, 2.49-, and 2.08-fold higher, respectively, compared to the control kept in darkness. These results demonstrate that the blue light receptor WC-2 functions as a positive regulator of GA biosynthesis in G. lingzhi, influencing the expression of genes involved in GA biosynthesis and asexual spore production, thereby advancing our understanding of the intricate regulatory network of GA biosynthesis. Full article

Welsh onion (Allium fistulosum L.), a globally significant vegetable, flavoring agent, and phytomedicine resource, has remained unavailable with established transient expression platforms for functional genomic investigations. To address this critical methodological limitation, we present systematically optimized protocols for both Agrobacterium-mediated hairy root transformation and protoplast transient expression systems, achieving significant advances in transformation efficiency for this species. Through systematic optimization of key parameters, including Agrobacterium rhizogenes (A. rhizogenes) strain selection (with Ar.Qual demonstrating superior performance), explant type efficacy, bacterial suspension optical density (OD₆₀₀ = 0.3), and acetosyringone induction concentration (100 μM), we established a highly efficient stem disc infection methodology, achieving 88.75% hairy root induction efficiency. Subsequent optimization of protoplast isolation protocols identified the optimal enzymatic digestion conditions: 6-h dark digestion of young leaves using 1.0% (w/v) Cellulase R-10, 0.7% (w/v) Macerozyme R-10, and 0.4 M mannitol, yielding 3.3 × 10⁶ viable protoplasts g⁻¹ FW with 90% viability. System functionality validation through PEG-mediated transient transformation demonstrated successful green fluorescent protein (GFP) reporter gene expression, confirmed by fluorescence microscopy. As the first documented transient expression platforms for Welsh onion, these protocols enable essential molecular investigations, including in planta promoter activity profiling, subcellular protein localization, and CRISPR-based genome-editing validation. This methodological breakthrough overcomes previous technical constraints in Welsh onion molecular biology, providing critical tools for accelerated gene functional characterization in this agriculturally important species. Full article

To systematically elucidate the chronological patterns of embryonic development and morphological changes in the larval and juvenile stages of Silurus glanis, and provide fundamental biological insights into this species, in this study, fertilized eggs were obtained through artificial spawning induction technology. After removing adhesiveness from fertilized eggs using trypsin, a detailed developmental study was conducted. The study systematically analyzed the chronological sequence of embryonic development and the morphological change patterns of larval and juvenile fish. The results showed the following: The fertilized eggs of S. glanis are yellow, spherical, and sticky, and the stickiness allows eggs to attach to spawning substrates, enhancing hatching success. The egg diameter after water absorption was (2.88 ± 0.13) mm. The embryonic development took 47 h and 55 min, with a total accumulated temperature of 1245.56 h degrees Celsius, the developmental process includes seven stages and twenty-six periods, namely the zygophase stage, cleavage stage, blastula stage, gastrula stage, neurula stage, organogenesis stage, and hatching stage. At a temperature of (26.0 ± 0.9) °C, the hatched individuals went through the pre-yolk sac larval stage, late larval stage, juvenile fry stage, and juvenile stage. In the pre-yolk sac larval stage, otoliths appeared in the bilateral otic vesicles, a pair of barbel primordia emerged under the mandible, a short and thin straight intestine formed in the abdominal cavity, and the oral fissure first appeared. In the late larval stage, the fin rays were initially formed, the intestine became thicker and longer, the oral fissure, anus, and cloaca were formed, and the larvae could float and start feeding on exogenous food. In the juvenile fry stage, the differentiation of various organs was basically complete, the nostrils became larger, and both the anal fin and caudal fin had dark black markings. In the juvenile stage, the maxillary barbels elongated, the mucus layer thickened on the body and back, the abdomen is light white, and it had the external morphological characteristics of an adult fish. By measuring and calculating the total length, body length, body height, and head length of S. glanis larvae and juveniles (0–40 days), the results showed that the growth characteristics conformed to the following fish growth formula: TL = 0.0141x² + 0.8096x + 8.2421 (R² = 0.9916), where x denotes days after hatching. This study has preliminarily mastered the chronological patterns of the embryonic development, growth, and formation of the morphological characteristics in larval and juvenile S. glanis, providing scientific data and laying a theoretical foundation for the division of early developmental stages, reproduction, hatching, and fry cultivation. Full article

Disruption of circadian rhythms by abnormal light exposure and reduced melatonin secretion has been linked to heightened pain sensitivity and opioid tolerance. This study evaluated how environmental light manipulation and exogenous melatonin supplementation influence pain perception and morphine tolerance in a rat model of neuropathic pain induced by partial sciatic nerve transection (PSNT). Rats were exposed to constant darkness, constant light, or a 12 h/12 h light–dark cycle for one week before PSNT surgery. Behavioral assays and continuous intrathecal (i.t.) infusion of morphine, melatonin, or their combination were conducted over a 7-day period beginning immediately after PSNT. On Day 7, after discontinued drugs infusion, an acute intrathecal morphine challenge (15 µg, i.t.) was administered to assess tolerance expression. Constant light suppressed melatonin levels, exacerbated pain behaviors, and accelerated morphine tolerance. In contrast, circadian-aligned lighting preserved melatonin rhythms and mitigated these effects. Melatonin co-infusion attenuated morphine tolerance and enhanced morphine analgesia. Reduced pro-inflammatory cytokine expression and increase anti-inflammatory cytokine IL-10 level and suppressed astrocyte activation were also observed by melatonin co-infusion during morphine tolerance induction. These findings highlight the potential of melatonin and circadian regulation in improving opioid efficacy and reduced morphine tolerance in managing neuropathic pain. Full article

Roses are one of the most essential ornamental flowers in the world. At present, traditional techniques such as cross breeding are mainly used in rose breeding. The inefficiency of the in vitro regeneration system has become the limiting step for the innovation and genetic improvement of rose germplasm resources. A tissue culture rapid propagation system of Rosa ‘Pompon Veranda’ was established using the stem segments with shoots as the initial experimental material. The results showed that the best disinfection method was to soak the explants in 75% ethanol for 1 min, and then soak them in 15% sodium hypochlorite solution for 15 min. The contamination rate was only about 6%. The best rooting medium for tissue culture seedlings was 1/2MS with 0.1 mg∙L⁻¹ NAA, and the rooting rate can reach around 95%. On this basis, calluses were induced by using leaflets of tissue-cultured seedlings as explants. The results showed that the optimal medium for inducing callus tissue was MS + 5.0 mg∙L⁻¹ 2,4-D, with an induction rate of 100%. The calluses were cultured in the medium of MS with 0.01 mg∙L⁻¹ NAA, 1.5 mg∙L⁻¹ TDZ and 0.1 mg∙L⁻¹ GA₃ for 12 days in the dark and then transferred to light conditions. The differentiation rate of callus was 10.87%. On the medium of MS with 0.5 mg∙L⁻¹ 6-BA, 0.004 mg∙L⁻¹ NAA and 0.1 mg∙L⁻¹ GA₃, the shoots could regenerate into whole plants. This study has established an in vitro regeneration system of R. ‘Pompon Veranda’, which is a key perquisite for the subsequent establishment of its genetic transformation system. Moreover, this method will also be an important reference for studies on quality traits such as floral scent and prickles of Rosa plants. Full article

An efficient in vitro propagation protocol for Eulophia bicallosa was developed using asymbiotic seed germination and protocorm proliferation. The effect of light on seed germination and development was evaluated on Vacin and Went (VW) medium under five conditions: darkness, white, green, red, and blue light for 24 weeks. Blue and red light significantly accelerated seed development, allowing progression to stage 5 within 24 weeks. For protocorm proliferation, six semi-solid culture media were tested. Half-strength Murashige and Skoog (½MS) medium yielded the best results after 8 weeks, producing the highest numbers of shoots (1.0), leaves (1.1), and roots (4.2) per protocorm, with 100% survival. The effects of organic additives were also evaluated using coconut water and potato extract. A combination of 200 mL L⁻¹ coconut water and 50 g L⁻¹ potato extract enhanced shoot formation (1.7 shoots), while 150 mL L⁻¹ coconut water with 50 g L⁻¹ potato extract increased both leaf (1.9) and root (8.8) numbers. The effects of cytokinins (benzyladenine (BA), kinetin (6-furfurylaminopurine), and thidiazuron (TDZ)) and auxins (indole-3-acetic acid (IAA), α-naphthalene acetic acid (NAA), indole-3-butyric acid (IBA), and 2,4-dichlorophenoxyacetic acid (2,4-D)) were investigated using ½MS medium supplemented with each plant growth regulator individually at concentrations of 0, 0.1, 0.5, 1.0, and 2.0 mg L⁻¹. Among the cytokinins, 0.1 mg L⁻¹ BA produced the highest survival rate (96%), while 1.0 mg L⁻¹ BA induced the greatest shoot formation (93%, 2.3 shoots). Among the auxins, 0.1 mg L⁻¹ IAA resulted in the highest survival (96%), and 1.0 mg L⁻¹ IAA significantly enhanced root induction (4.2 roots per protocorm). Acclimatization in pots containing a 1:1:1 (v/v) mixture of pumice, sand, and soil resulted in 100% survival. This protocol provides a reliable and effective approach for the mass propagation and ex situ conservation of E. bicallosa. Full article

Bombax ceiba is an important medicinal and ornamental tree widely distributed in tropical and subtropical areas. However, its seeds lose viability rapidly after harvest, which has created hurdles in large-scale propagation. Here, we describe the development of a rapid and efficient de novo organogenesis system for Bombax ceiba, incorporating both indirect and direct regeneration pathways. The optimal basal medium used throughout the protocol was ½ MS supplemented with 30 g/L glucose, with all cultures maintained at 26–28 °C. For the indirect pathway, callus was induced from both ends of each hypocotyl on basal medium supplemented with 0.2 mg·L⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg·L⁻¹ 6-Benzylaminopurine (6-BA) under dark conditions. The induced calluses were subsequently differentiated into adventitious shoots on basal media containing 0.5 mg·L⁻¹ Indole-3-butyric acid (IBA), 0.15 mg·L⁻¹ Kinetin (KIN), and 1 mg·L⁻¹ 6-BA under a 16 h photoperiod, resulting in a callus induction rate of 140% and a differentiation rate of 51%. For the direct regeneration pathway, shoot buds cultured on medium with 0.5 mg·L⁻¹ IBA and 1 mg·L⁻¹ 6-BA achieved a 100% sprouting rate with a regeneration coefficient of approximately 3.2. The regenerated adventitious shoots rooted successfully on medium supplemented with 0.5 mg·L⁻¹ Naphthylacetic acid (NAA) and were acclimatized under greenhouse conditions to produce viable plantlets. This regeneration system efficiently utilizes sterile seedling explants, is not limited by seasonal or environmental factors, and significantly improves the propagation efficiency of Bombax ceiba. These optimized micropropagation methods also provide a robust platform for future genetic transformation studies using hypocotyls and shoot buds as explants. Full article

Fucoxanthin, a carotenoid with notable pharmaceutical potential, has attracted significant attention due to its efficient accumulation in marine microalgae and the importance of optimizing its induction conditions. In this study, Phaeodactylum tricornutum was employed as a model organism to screen optimal conditions for fucoxanthin accumulation using a three-factor, four-level orthogonal design. Furthermore, the underlying mechanisms related to photosynthetic physiology and gene regulation were explored. The results revealed that both glycine (Gly) and light intensity significantly enhanced fucoxanthin content (p < 0.05). The optimal condition (Combination C: 0.50 g L⁻¹ Gly, 36 μmol photons·m⁻²·s⁻¹, 12 h light/12 h dark) yielded a fucoxanthin content of 0.87 μg g⁻¹, representing a 35% increase compared to the control. Meanwhile, Combination p (0.50 g L⁻¹ Gly, 36 μmol photons·m⁻²·s⁻¹, 24 h light/0 h dark) significantly improved cell density (5.11 × 10⁶ cells mL⁻¹; +18%) and fucoxanthin yield (4.10 μg L⁻¹; +47%). Analysis of photosynthetic parameters demonstrated that the non-photochemical quenching coefficient (NPQ) was suppressed. Gene expression profiling showed that Combination C upregulated photosynthetic genes (psbA, rbcL, rbcS) by up to 2.36-fold, while Combination P notably upregulated fcpb (7.59-fold), crtiso, and pds. Principal component analysis identified that rbcS and pds are key regulatory genes. These findings demonstrate that Gly, light intensity, and photoperiod synergistically regulate the expression of genes involved in photosynthesis and carotenoid biosynthesis, thereby promoting fucoxanthin accumulation. This work provides valuable insights and a theoretical basis for optimizing fucoxanthin production in support of marine drug development. Full article

Miscanthus × giganteus (Greef and Deuter ex Hodkinson and Renvoize) is a perennial, rhizomatous grass that has gained significant attention as an industrial crop, particularly as a bioenergy feedstock. It is a natural interspecific hybrid with 57 chromosomes (2n = 3x = 57). Due to its sterility, M. × giganteus has limited genetic variability, making traditional breeding methods ineffective for its improvement. Consequently, alternative approaches are being explored to enhance its cultivation and utility. The study aimed to investigate the potential for M. × giganteus plant regeneration through ovary and flower bud cultures. Indirect in vitro regeneration of M. × giganteus plants was successfully achieved using flower bud cultures. Embryogenic-like callus was derived from explants originating from inflorescences that had undergone a four-day pretreating at 10 °C. The most effective medium for callus induction was a modified MS medium supplemented with 5 mg·dm⁻³ dicamba, 0.2 mg dm⁻³ 6-benzylaminopurine, 30 g dm⁻³ sucrose, and solidified with 8 g dm⁻³ agar or agarose. The optimal conditions for callus induction were achieved by culturing in the dark. The regenerated plants exhibited the characteristic chromosome number of the species, confirming that the regenerants did not develop from embryo sac cells. In contrast, ovary culture failed to produce callus or regenerated plants, highlighting its ineffectiveness for M. × giganteus regeneration. These findings underscore the potential of flower bud culture as a successful in vitro regeneration method while demonstrating the limitations of ovary culture for this species. Full article

In vitro anther culture is a technique used to produce haploid plants when regenerating varieties with specific traits. To generate haploid plants with preferred characteristics, an anther culture technique was established for Lilium longiflorum “Show Up”. Morphological characteristics were recorded, including the flower bud length and anther color corresponding to different stages of microspore development. The effects of different flower bud lengths, various concentrations of exogenous plant growth regulators (PGRs), low-temperature pretreatment at 4 °C, and incubation under dark conditions on the induction of callus formation were studied. When the flower buds were 2.2–2.4 cm in length and the microspores were in the mononuclear development phase, callus induction reached the highest rate (15.6%). Callus was not induced when the PGRs 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (KT) were added separately to the growth medium, but the highest callus induction rate occurred when anthers were cultured on the medium containing 2,4-D (0.75–1.0 mg/L) and KT (4 mg/L). The low-temperature pretreatment significantly enhanced the induction rate of anthers, but prolonged low-temperature pretreatment reduced the induction rate. The optimal period of cultivation in darkness was 6 d. After 15 days of cultivation, the number of swollen anthers was recorded, and these were transferred onto the differentiation medium Murashige and Skoog (MS) + 1-naphthaleneacetic acid (NAA) (2.0 mg/L), sucrose (30 g/L), and agar (7 g/L) at pH 5.8, whereon 100% differentiation was recorded. Overall, 14 regenerated lines were obtained by in vitro anther culture. Chromosome ploidy was determined by counting chromosomes in the root tips of ten regenerated plants, and four were found to be haploids. This study lays the foundation for anther culture in lilies to shorten the breeding cycle, improve selection efficiency, facilitate efficient genetic transformation, and enable the effective production of both haploid and double-haploid plants. Full article

This study presents the optimal light spectrum and medium composition for the in vitro induction of adventitious bulbs in T. tarda Stapf. Bulb scales, used as explants, were cultivated on 100% MS solid media with 3% or 6% sucrose and 0 or 0.5 µM BAP (6-benzyl-aminopurine) under a 16 h photoperiod and different fluorescent light treatments (white, red, blue). Darkness was used as a control. The cultures were maintained at 20 ± 2 °C for 12 weeks. The obtained results revealed that white light combined with 6% sucrose yielded the highest adventitious bulb formation, with an average of 12.1 ± 1.3 bulbs per explant. The bulbs formed directly on the surface of the explants. Red light combined with 3% sucrose and 0.5 µM BAP completely inhibited bulb formation, while darkness promoted leaf development, with a maximum of 1.3 ± 0.1 leaves per bulb, under darkness on medium with 6% sucrose. The addition of BAP, in most cases, was essential for root formation, with a maximum of 2.9 ± 0.6 roots per bulb under the influence of white light. This study demonstrates that it is possible to obtain an effective and efficient method for T. tarda propagation from bulb scales treated with 6% sucrose under white light condition. This method offers the potential for the commercial cultivation of this ornamental species. Full article

The tea plant (Camellia sinensis) is an economically important crop that plays an important role not only in the beverage industry but also in the pharmaceutical industry. The environment has a great influence on the quality of the tea plant. Heat shock factors (Hsfs) are transcriptional regulators that control the plant response to adversity. However, only a limited number of studies have reported the Hsf gene in Camellia sinensis, and most of these reports involve high-temperature, drought, and salt stress. Research on light, dark, and cold stress is limited. In this study, 22 CsHsf genes were obtained by whole genome sequencing and found to be located on 11 chromosomes. In addition, the gene structure, protein motif, and phylogeny were studied. We classified the genes into three major subfamilies: CsHsfA, CsHsfB, and CsHsfC. Interestingly, we found that there was more alignment between CsHsf and Hsf genes in dicotyledons, including Arabidopsis thaliana and Solanum lycopersicum, than in the monocotyledon Oryza sativa. The expression of many CsHsf genes was affected by low-temperature, light, and dark abiotic stresses. Notably, CsHsf15 and CsHsf16 showed high induction rates under both light and cold stress, and both genes carried cis-acting elements associated with light and low-temperature responses. These results lay a solid groundwork for further investigations into the involvement of CsHsf genes in the response of Camellia sinensis to abiotic stresses. Full article

Nanoparticles play an important role in plant response to abiotic stresses including salt stress. In this study, the physiological and histological responses of CuO, ZnO, and CaO nanoparticle (NP) applications on callus tissues developed from two alfalfa lines (Erzurum and Muş) exposed to salt (NaCl) stress were evaluated. The NPs were synthesized from the extracts obtained from healthy walnut shells using the green synthesis approach and then characterized by Scanning Electron Microscopy (SEM) and X-ray diffraction analysis (XRD). The leaf explants were placed in an MS medium containing 4 mg L⁻¹ 2,4-D (2,4-dichlorophenoxyacetic acid), 50 mM NaCl, and 0.8 ppm of NPs for 1 month in the dark. CaO NP is determined to be more effective than CuO and ZnO in callus induction from leaf explants. Malondialdehyde (MDA) content was higher in the callus treated with 0.8 ppm CuO NP + 50 mM NaCl compared to other treatments. The callus induction stage, without salt treatments, showed the best results with 0.8 ppm CaO NPs for both H₂O₂ levels and peroxidase (POX) activity compared to the other NPs. The highest protein rate was obtained from the callus induction stage and callus formation stage after 50 mM treatment NaCl with 0.8 ppm CuO. The LCSM results displayed, under in vitro conditions, that the treatment of NPs can greatly suppress the negative effects of salt stress on calli samples. SEM analysis supported the results obtained by laser scanning confocal microscopy (LSCM) analysis. Our findings suggest that CuO, CaO, and ZnO NPs can offer a simple and effective method to protect alfalfa callus from NaCl stress severity. Furthermore, these NPs, particularly CaO, hold potential for broader application and should be evaluated under various abiotic conditions beyond salt stress. Full article