Search Results (22)

The pokeweed (Phytolacca americana L.) plant is native to North America and contains bioactive compounds with medicinal potential, particularly phenolics and saponins. This study enhanced the production of secondary metabolites in pokeweed callus cultures using sodium chloride (NaCl) and polyethylene glycol (PEG) as elicitors under aseptic conditions. Pokeweed seeds were cultured on Murashige and Skoog (MS) medium for 8 weeks. Fully expanded leaves from the second to third position from the shoot were excised and induced to form calli on MS medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) for 5 weeks. Fully developed calli were elicited with PEG6000 at concentrations of 0, 1.25, 2.5, and 5% (w/v) in combination with NaCl at concentrations of 0, 100, 200, and 300 mM for 15 days. Callus growth was recorded, followed by drying and extraction using methanol (MeOH) for biochemical analysis. Calli elicited with 2.5% PEG and 300 mM NaCl exhibited the highest total phenolic content (TPC) (21.063 µg GAE/mg DW) and total flavonoid content (TFC) (1.927 µg QUE/mg DW). The highest antioxidant activities determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and ferric ion reducing antioxidant potential (FRAP) assays were 0.998, 1.574, and 0.998 µg TE/mg DW, respectively. The elicitation of pokeweed calli with 300 mM NaCl yielded the highest amount of Esculentoside A (EsA) (15.753 µg/mg DW). All the elicitor treatments significantly enhanced metabolite accumulation compared to the control group (p < 0.05). The findings indicated that elicitation with PEG and NaCl effectively enhanced the production of secondary metabolites in P. americana callus cultures. This study offers a promising alternative approach for utilizing P. americana in pharmaceutical and medicinal applications. Full article

Background: Common variable immunodeficiency (CVID), the most prevalent symptomatic inborn error of immunity, involves impaired B-cell differentiation and antibody production, causing recurrent infections and the need for life-long immunoglobulin replacement therapy. Methods: This study evaluated the in vitro differentiation of memory B-cells (MBCs) into antibody-secreting cells (ASCs) in CVID patients. Peripheral blood mononuclear cells from 13 CVID patients and 10 healthy controls were stimulated using two protocols: (I) Staphylococcus aureus Cowan Strain I, Pokeweed mitogen, and CpG, or (II) a T-cell-dependent approach using CD40 ligand, interleukin-21, and CpG. B-cell subpopulations were analyzed by flow cytometry, ASC differentiation using ELISpot, and antibody levels in supernatants by ELISA. Results: Despite severely restricted in vivo antibody production, MBCs from all 13 CVID patients differentiated into IgG and IgM ASCs under adequate in vitro stimulation. Protocol II, mimicking T-cell help, was more effective than protocol I. As expected, the patients exhibited reduced class-switched MBCs ex vivo, but the MBCs differentiated and proliferated to an extent similar to those in healthy controls. IgA secretion remained significantly impaired post-stimulation. Specific IgG antibodies against tetanus toxoid and Streptococcus pneumoniae were detected in the patient supernatants, while no double-stranded DNA autoantibodies emerged after in vitro stimulation. Conclusions: These findings indicate that the MBCs of most patients retain functional B-cell differentiation and antigen-specific IgG secretion under T-cell dependent stimulation, though IgA secretion remains impaired. Tailored stimulation protocols could deepen our understanding of how to restore MBC formation in CVID patients in vivo. This methodology provides a platform to investigate antigen-specific functional memory responses like post-vaccination. Full article

The apoplast is often the first point of contact between plant cells and invading pathogens, serving as an important site for defense signaling. Pokeweed antiviral protein (PAP), a ribosome-inactivating protein from Phytolacca americana (pokeweed), is localized to the apoplast and is hypothesized to accompany a pathogen to the cytosol, where it would inactivate host ribosomes to prevent pathogen spread. However, it is not known whether PAP interacts with other proteins in the apoplast. In this study, we identified Phytolacca americana cysteine protease 1 (PaCP1), an extracellular cysteine protease, as a novel PAP interactor. Sequence and structural analyses classified PaCP1 as a member of the C1A subfamily of papain-like cysteine proteases. Immunoprecipitation, mass spectrometry, and yeast two-hybrid analysis showed that PAP specifically binds the mature, active form of PaCP1. Curiously, PaCP1 cleaves PAP at its N- and C-termini, generating peptides that enhance MAPK phosphorylation in pokeweed leaves, indicating their potential role in stress signaling. PaCP1 processing of PAP to generate bioactive peptides diversifies the function of a ribosome-inactivating protein beyond its canonical inhibition of translation. Our findings present a novel extracellular role for PAP and advance our understanding of how protein interactions in the apoplast contribute to plant immune responses. Full article

In response to consumer demand for more sustainable and health-conscious products, the food sector is increasingly shifting towards the use of natural additives. Pokeweed (Phytolacca americana L.) is a medicinal plant that contains valuable biologically active compounds, including betacyanins, which serve as its red pigments, along with phenolic acids, flavonoids, polyphenolic compounds, and others. Phytolacca americana (P. americana) is a plant renowned for its bioactive compounds, which exhibit anti-inflammatory, anti-mutagenic, antioxidant, anticancer, and antibacterial properties. This study investigates the potential of betalain pigments extracted from the berries of P. americana as a natural colorant for cheese formulation. The impact of these pigments on the color attributes, sensory qualities, and physicochemical and phytochemical composition of the cheeses was systematically evaluated. The Phytolacca americana (PA) powder demonstrated significant levels of total polyphenols (111.95 ± 1.60 mg GAE/g dw) and antioxidant activity (21.67 ± 0.19 µmol TE/g dw). The incorporation of PA powder increased the physicochemical and phytochemical contents and antioxidant activity in the final product (4.40 ± 0.22 µmol TE/g dw for CPAP1 and 6.11 ± 0.22 µmol TE/g dw for CPAP2). The sensory study revealed that the PA-supplemented cheeses were acceptable. The enhanced cheeses present a distinctive color profile, attracting health-conscious consumers looking for innovative dairy products. The study concludes that PA powder can effectively enhance cheese, producing a phytochemical-enriched product that appeals to health-conscious consumers. Full article

Wheat (Triticum aestivum L.) is one of the most agriculturally and economically important crops in the world. Wheat fungal diseases are becoming more severe and frequent due to global climate change, threatening wheat yields and security. While fungal diseases such as fusarium head blight, stripe rust, and powdery mildew have been extensively studied, the newly emerged fungal pathogens in wheat are still under-researched. In May 2023, black mold symptoms were observed on wheat spikes in Xinxiang City, Henan Province, China. However, the causal agent of this disease was not known. We employed a combination of morphological examination and molecular techniques to identify the pathogen. The internal transcribed spacer (ITS) region, translation elongation factor 1-alpha (tef1), and actin (act) genes of the fungus were partially sequenced (accession no. OR186209, PQ271633 and PQ271632) and showed 99.59–100% identity with the previously reported Cladosporium cladosporioides, which affects wheat, pokeweed, and black-eyed pea. The pathogenicity of this fungus was confirmed by fulfilling Koch’s postulates. Through a rigorous screening process, we found Simplicillium aogashimaense, Trichothecium roseum, and Bacillus velezensis as effective biocontrol agents, with B. velezensis demonstrating the most potent antagonistic activity against the Cladosporium mold. This discovery showed the potential of B. velezensis as a biocontrol agent for wheat disease management. The findings underscore the importance of the present study in advancing the control of this disease. Full article

Forests are important ecosystems that face threats from climate change and global environmental shifts, with invasive alien plant species being a significant concern. Some of these invasive species have already become established, while others are in the process of naturalisation. Although forests are a relatively stable ecosystem, extreme weather events increase their vulnerability to change, and clearings left after natural disturbances are particularly susceptible to invasion by alien plant species (IAPS). We created risk maps of two species that have spread rapidly in the last decade: American pokeweed (Phytolacca americana) and the tree of heaven (Ailanthus altissima). We prepared a generalised linear model based on the occurrence data collected within the LIFE ARTEMIS project. Eleven environmental variables were used to determine habitat characteristics. We constructed two models for each species: one covering the entirety of Slovenia and the other specifically for the forested areas in Slovenia, with the latter incorporating forest-specific variables (such as forest sanitation felling and monocultures). We observed the presence of both species at lower altitudes and in close proximity to water sources. American pokeweed tends to occur nearer to railways, while the presence of the tree of heaven is associated with areas lacking carbonate parent material and influenced by land use patterns. In forested areas, the occurrence of American pokeweed is influenced by forest habitat characteristics, such as disturbances caused by extreme weather events or the prevalence of Norway spruce monocultures. In contrast, the occurrence of the tree of heaven is influenced by more general environmental variables, such as altitude and proximity to railways. Consequently, we have generated risk maps for the entirety of Slovenia and separately for forested areas, both of which indicate similar levels of risk, particularly for the tree of heaven. The risk map for American pokeweed highlights numerous vulnerable areas, especially forest edges, which are highly susceptible to invasion. Furthermore, there is a higher likelihood of this species occurring in areas that have undergone sanitation felling. This study suggests that the production of risk maps of IAPS could be improved by focussing on habitat types and taking into account habitat-specific variables. This approach could enhance the early detection and management of these invasive species. Full article

Manganese (Mn) exclusion in the Mn hyperaccumulator pokeweed (Phytolacca americana L.) was investigated. Hydroponic experiments were carried out to observe the responses of pokeweeds continually exposed to high levels of Mn. In this study, crystals were observed to appear firstly on the root hair, and soon after, more crystals appeared on the root surface, and crystals of Mn phosphate were observed to appear on the root surface in a time sequence negatively correlated with the number of leaves treated with 5 mM Mn. Crystals were identified via phase analysis of X-ray diffraction and element analysis, and these white insoluble crystals were identified using XRD to be Mn phosphate, with the molecular formula (Mn,Fe)₃(PO₄)₂·4H₂O. The nutrient solution pH increased from 4.5 to about 5.6 before the crystals appeared. Mn phosphate crystals appeared in all solutions except those without phosphate and emerged earlier in the solutions containing no Fe. Compared with control group, pokeweed accumulated much more Mn in the leaves when treated without phosphate or Fe. The present study suggests that pokeweed can exclude Mn by means of rhizosphere precipitation by phosphate to form Mn phosphate crystals that accumulate on the root surface. Although the detailed mechanism requires further investigation, this study provides the first direct evidence of a novel strategy to inhibit Mn uptake in the roots of a hyperaccumulator in a P-enriched environment. Full article

Mathematical models are becoming indispensable tools to explore the complexities of biological systems at cellular levels. We present a model to explore the baseline immune cell interactions for in vitro polyclonal antibody synthesis via B-cells regulated by helper and regulatory T-cells. The model incorporates interactions of antigen-presenting cells, T-cells, regulatory T-cells, and B-cells with each other and predicts time-dependent trajectories of these cells and antibody synthesis stimulated by pokeweed mitogen. We used an ordinary differential equation-based approach to simulate the dynamic changes in the cells and cytokines numbers due to the cellular and humoral response to pokeweed mitogen stimulation. The parameters of the ordinary differential equations model are determined to yield a normal immune response as observed in the pokeweed mitogen-stimulated in vitro antibody synthesis via normal T, B, and antigen-presenting cells. The dose effects of antigen load and basal values of regulatory T-cells on the profiles of various immune response variables are also evaluated. Full article

Food packaging films were prepared by using guar gum/polyvinyl alcohol (GP) as the film matrix, 2% Ag nanoparticles (AgNPs) as reinforcing filler and antimicrobial agent, and 1%, 2% and 3% pokeweed betacyanins (PB) as the colorant and antioxidant agent. The structures and color-changing, barrier, mechanical, thermal and antioxidant/antibacterial properties of different films were measured. The results show that the PB were pH-sensitive pigments with pink, purple and yellow colors at pH 3–8, pH 9–11 and pH 12, respectively. PB improved the compatibility of guar gum and polyvinyl alcohol through hydrogen bonds. The films with PB showed a color-changing capacity under ammonia vapor and good color stability in chilled storage. AgNPs and PB elevated the barrier capacity of GP film to light, water vapor and oxygen gas. Meanwhile, AgNPs and PB improved the stiffness, thermal stability and antioxidant/antibacterial activity of GP film. The film with AgNPs and 3% PB showed the highest barrier capacity, stiffness, thermal stability and antioxidant/antimicrobial activity. In shrimp spoilage test, the films with AgNPs and 2% and 3% PB indicated shrimp freshness through film color changes. The results reveal the potential use of the prepared films in active and smart packaging. Full article

Oedema disease (OD) in piglets is one of the most important pathologies, as it causes significant losses due to the high mortality because of the Shiga toxin family, which produces Escherichia coli (STEC) strains. The main toxin responsible for the characteristic pathologies in pigs is Shiga toxin 2 subtype e (Stx2e). Moreover, there is growing evidence that Stx’s family of toxins also targets immune cells. Therefore, this study evaluated the effect of different concentrations of Stx2e on porcine immune cells. Porcine peripheral blood mononuclear cells were pre-incubated with Stx2e, at three different concentrations (final concentrations of 10, 500, and 5000 CD50/mL) and with a negative control group. Cells were then stimulated with polyclonal mitogens: concanavalin A, phytohemagglutinin, pokeweed mitogen, or lipopolysaccharides. Cell proliferation was assessed by BrdU (or EdU) incorporation into newly created DNA. The activation of the lymphocyte subsets was assessed by the detection of CD25, using flow cytometry. The toxin significantly decreased mitogen-driven proliferation activity, and the effect was partially dose-dependent, with a significant impact on both T and B populations. The percentage of CD25+ cells was slightly lower in the presence of Stx2e in all the defined T cell subpopulations (CD4+, CD8+, and γδTCR+)—in a dose-dependent manner. B cells seemed to be the most affected populations. The negative effects of different concentrations of Stx2e on the immune cells in this study may explain the negative impact of the subclinical course of OD. Full article

Phytolacca americana and P. acinosa are alien plant species in Europe. The former is considered invasive and more widespread. In order to develop effective and safe eradication and plant disposal methods, the present research focused on the seed germination of the two species. Fruits of different ripeness of both species were collected (fresh and dry seeds within and without pericarp), after which both the germination and maturation were tested. We also tested the continued maturing of fruits on cut plants and observed the development of fruits on whole plants with a cut taproot (in addition to when only the upper part of the stem with fruit racemes was cut off). In general, the seeds germinated from all stages of fruit ripeness, although the germination of dry seeds was better compared to fresh seeds. P. americana’s seeds germinated better and the fruit ripening on cut plants was also more successful compared to P. acinosa. These results could partly explain the invasive success of P. americana. According to our results, removing all fruiting plants from the eradication site is crucial regardless of the fruit development stage. Full article

In anaerobic soil disinfestation (ASD), soil amended with a carbon source undergoes anaerobic conditions accompanied by changes in microbial community composition and an increase in the concentration of organic acids, primarily acetic and butyric acids, and gases that are deleterious to plant pathogens, insects, and potentially to weeds. The purpose of this study was to explore the efficacy of ASD with different carbon sources on inactivation of propagules of a variety of weed species. Germination and viability of propagules of common lambsquarters (Chenopodium album L.), black nightshade (Solanum nigrum L.), yellow nutsedge (Cyperus esculentus L.), common pokeweed (Phytolacca decandra L.), barnyardgrass (Echinochloa crus-galli L.), dandelion (Taraxacum officinale (Weber)), and redroot pigweed (Amaranthus retroflexus L.) were tested using different rates of four carbon sources: molasses, wheat bran, mustard greens biomass, and raw chicken manure. Wheat bran was the most effective carbon source for inactivation of all weed propagules, followed by molasses and mustard greens biomass. Carbon sources were mixed with soil, which was irrigated to saturation and covered with plastic to promote anaerobic conditions for three weeks. Chicken manure inactivated all the tested species except A. retroflexus compared to the anaerobic control. Rates of carbon sources applied in soil were weakly but significantly correlated (r > 0.42 and p < 0.04) with weed seed mortality or germination inhibition (E. crus-galli) for all the tested carbon sources and weed species except for chicken manure for C. album, A. retroflexus, and S. nigrum. Laboratory tests to confirm organic acid toxicity showed that 1000 µL L⁻¹ acetic plus 1000 µL L⁻¹ butyric acids for at least 60 h inhibited of sprouting of C. esculentus tubers and E. crusgalli seeds, whereas 120 h or exposure was required to inhibit C. album seed germination. Anaerobic soil disinfestation effectively suppressed weed seed germination and viability when wheat bran, molasses and mustard greens biomass were used as carbon sources. This approach could be adopted for weed management in organic systems or any farming system where priority is given to environmentally sustainable practices. Full article

We performed a detailed HPLC-MSn analysis of the phenolic compounds from the extracts of two invasive alien plant species (IAPS): Japanese knotweed (Fallopia japonica (Houtt.) Ronse Decr.) and American pokeweed (Phytolacca americana L.). The major phenolic groups were hydroxycinnamic acids and flavanols in Japanese knotweed (J. knotweed) and flavonols, hydroxycinnamic acids, and stilbenes in American pokeweed (A. pokeweed). We investigated the influence of solvent type and extraction time on the extraction efficiency of the phenolic compounds. The solvent 80% methanol had a higher polyphenolic extraction efficiency than water, since 14.5 times more flavonols and 2.3 times more stilbenes were extracted from J. knotweed and 5.2 times more flavonols and 2.6 times more stilbenes were extracted from A. pokeweed. In contrast, with water, we obtained a 52% higher hydroxycinnamic acids (HCA) content from J. knotweed. Hydroxycinnamic acids were best extracted in water after 24 h, flavanols after 12 h, stilbenes between 12 and 24 h, and flavonol glycosides after 48 h of extraction. We also tested the allelopathic effect of the aqueous extract of A. pokeweed and J. knotweed on seed germination and shoot and root growth of perennial ryegrass. The results showed that the water extract of J. knotweed resulted in 38 to 48% lower seed germination of perennial ryegrass, and the extract of A. pokeweed resulted in 83 to 90% lower seed germination. The phytotoxic effect of the extract of J. knotweed and A. pokeweed was also reflected in a characteristic reduced growth of shoots and roots of perennial ryegrass. The phytotoxic action of IAPS could also be applied for beneficial purposes, since this would be an effective strategy for their control and a reduction of their spread in the environment. Full article

We recently identified a deviant bovine immune phenotype characterized by hyperproliferation of lymphocytes after polyclonal stimulation. This phenotype was first discovered in dams that responded to PregSure BVD vaccination by producing pathological antibodies, triggering the fatal disease “bovine neonatal pancytopenia” in calves. The aim of the study was to gain deeper insights into molecular processes occurring in lymphocytes of immune phenotypes and the effect on their secretome after immune stimulation. Two discovery proteomic experiments were performed with unstimulated and Pokeweed Mitogen (PWM) stimulated lymphocytes, using label-free LC-MS/MS. In lymphocytes, 2447 proteins were quantified, and 1204 proteins were quantified in the secretome. Quantitative proteome analysis of immune deviant and control samples after PWM stimulation revealed clear differences. The increase in abundance of IL17A, IL17F, IL8, CCL5, LRRC59, and CLIC4 was higher in controls through mitogenic stimulation. In contrast, the abundance of IFNγ, IL2, IL2RA, CD83, and CD200 increased significantly more in immune deviant lymphocytes. Additional pathway enrichment analysis of differentially secreted proteins also yielded fundamental differences between the immune phenotypes. Our study provides a comprehensive dataset, which gives novel insights into proteome changes of lymphocytes from different bovine immune phenotypes. These differences point to the development of diverse immune responses of bovine immune phenotypes after immune stimulation. Full article

Drosophila suzukii, an economically important pest of small and thin-skinned fruits, has caused annual crop losses up to 20% in the state of Georgia’s multimillion-dollar blueberry industry. The known host range of D. suzukii is large, yet the breadth of uncultivated and wild plants that can serve as alternative hosts in the southeastern United States is still not fully understood. Establishing comprehensive lists of non-crop D. suzukii hosts in woodlands near blueberry production will assist in the creation of more sustainable integrated pest management (IPM) strategies. Objectives of this study were to determine viability of wild fruiting plant species to this pest based on survivorship to adulthood and assess D. suzukii short-range preference between cultivated blueberries and wild fruit. Laboratory choice and no-choice assays were performed to determine if D. suzukii could complete its development on wild fruits sampled from the field. Results from our no-choice assays indicated that multiple species of wild fruits surveyed in Georgia were viable D. suzukii hosts including blackberry species, deerberry, hillside blueberry, common pokeweed, beautyberry, elderberry, evergreen blueberry, and large gallberry. Yet, none of these hosts were preferred by adult female D. suzukii as ovipositional substrates when compared to cultivated blueberries. However, these uncultivated species have the potential to sustain D. suzukii populations pre- and post-harvest season. This information can help farmers do more targeted management of these viable alternative hosts from wooded areas surrounding blueberry fields in order to minimize D. suzukii populations. Full article