Search Results (43)

Oxidative stress is key in immune defense against fungal infections, such as those caused by Sporothrix schenckii, the dimorphic fungus responsible for sporotrichosis. Phagocytic cells utilize oxidative stress as a crucial mechanism to control pathogen spread. During S. schenckii infection, phagocytic cells recognize pathogen-associated molecular patterns (PAMPs) on their surface through conserved transmembrane or soluble receptors, known as pattern recognition receptors (PRRs). This recognition triggers a cascade of immune responses, including the generation reactive oxygen species (ROS) essential for pathogen elimination. However, S. schenckii has developed sophisticated mechanisms to evade and counteract this response, contributing to its persistence in the host. These mechanisms include the production of antioxidant enzymes, alterations to its cell wall (CW), and the production of melanin, which helps neutralize oxidative stress. In addition, S. schenckii modulates the production of other proteins, such as moonlighting proteins, suggested to have roles in immune evasion and stress response, helping its survival in the host. These strategies, along with the modulation of gene expression, allow the fungus to survive and persist inside the immune system’s hostile environment, facilitating the progression of the infection. Understanding these interactions between phagocytic cells and S. schenckii is key to developing more effective therapeutic strategies to combat sporotrichosis. Full article

The gas-phase delivery of lignin into the hot zone of cw-CO₂ laser-powered homogeneous pyrolysis (LPHP) reactor under “wall-less” conditions led to the breakdown of lignin macromolecules into neutral oligomers and paramagnetic fragments deposited onto the reactor cell walls. The formation of PAHs was observed during the defragmentation of lignin, accelerated with increased laser power. Remarkably, no phenolic compounds were detected among lignin fragments—intermediate radicals and neutral oligomers. It is concluded that the PAH and soot-like conjugated particulates are formed in the hot zone of the LPHP reactor, resembling the high-temperature combustion processes. The key role of the resonantly stabilized radicals in the formation of low-molecular-weight PAHs is outlined. An alternative pathway is proposed for the generation of PAH involving the formation of cyclopentadienyl radical precursors (CPDa) that are adsorbed onto or trapped within lignin macromolecules. Full article

Previous studies in the Salado River Basin (Argentina) demonstrated that the introduced forage species, Lotus tenuis Waldst. & Kit. ex Wild. (Fabaceae), possesses high tolerance to abiotic stresses—including flooding, alkalinity, salinity, and drought. The efficient biological fixation of nitrogen in a region with a scarce presence of native legumes is one of its advantages. Despite these qualities, a year-long characterization of cell wall (CW) polysaccharides in Lotus tenuis and their relationship with the high nutritional quality is missing. In this study, seasonal parametric investigations of L. tenuis, regarding its photosynthetic and ionic status, modifications in CW composition, and concomitant nutritional quality, were performed. Our results demonstrate the high plant digestibility and protein content of this legume, even in summer, when most accompanying species reduce their forage quality. Regarding gas production kinetics (in vitro production is a proxy for the animal rumen’s output), spring biomass had the highest values. The CW material yields are similar throughout the year, but with differences in polysaccharide composition. In summer and winter, pectins predominate, while in the regrowth periods (spring and autumn), pectins and β-glucans are found in similar amounts. This work confirms that Lotus tenuis can help optimize grassland productivity in challenging mesophytic terrains to increase livestock productivity through environmentally friendly services. Full article

Background: Tomatoes are self-pollinating plants, and successful fruit set depends on the production of functional pollen within the same flower. Our previous studies have shown that the ‘Black Vernissage’ tomato variety exhibits greater resilience to heat stress in terms of pollen productivity compared to the ‘Micro-Tom’ variety. Pollen productivity is determined by meiotic activity during microsporogenesis and the development of free microspores during gametogenesis. This study focused on identifying heat stress (HS)-induced proteomes in pollen mother cells (PMCs) and microspores. Methods: Tomato plants were grown under two temperature conditions: 26 °C (non-heat-treated control) and 37 °C (heat-treated). Homogeneous cell samples of meiotic PMCs (prior to the tetrad stage) and free microspores were collected using laser capture microdissection (LCM). The heat-induced proteomes were identified using tandem mass tag (TMT)–quantitative proteomics analysis. Results: The enrichment of the meiotic cell cycle in PMCs and the pre-mitotic process in free microspores confirmed the correlation between proteome expression and developmental stage. Under HS, PMCs in both tomato varieties were enriched with heat shock proteins (HSPs). However, the ‘Black Vernissage’ variety exhibited a greater diversity of HSP species and a higher level of enrichment compared to the ‘Micro-Tom’ variety. Additionally, several proteins involved in gene expression and protein translation were downregulated in PMCs and microspores of both varieties. In the PMC proteomes, the relative abundance of proteins showed no significant differences between the two varieties under normal conditions, with very few exceptions. However, HS induced significant differential expression both within and between the varieties. More importantly, these heat-induced differentially abundant proteins (DAPs) in PMCs are directly involved in meiotic cell division, including the meiosis-specific protein ASY3 (Solyc01g079080), the cell division protein kinase 2 (Solyc11g070140), COP9 signalosome complex subunit 1 (Solyc01g091650), the kinetochore protein ndc80 (Solyc01g104570), MORC family CW-type zinc finger 3 (Solyc02g084700), and several HSPs that function in protecting the fidelity of the meiotic processes, including the DNAJ chaperone (Solyc04g009770, Solyc05g055160), chaperone protein htpG (Solyc04g081570), and class I and class II HSPs. In the microspores, most of the HS-induced DAPs were consistently observed across both varieties, with only a few proteins showing significant differences between them under heat stress. These HS-induced DAPs include proteases, antioxidant proteins, and proteins related to cell wall remodeling and the generation of pollen exine. Conclusions: HS induced more dynamic proteomic changes in meiotic PMCs compared to microspores, and the inter-varietal differences in the PMC proteomes align with the effects of HS on pollen productivity observed in the two varieties. This research highlights the importance of the cell-type-specific proteomics approach in identifying the molecular mechanisms that are critical for the pollen developmental process under elevated temperature conditions. Full article

The plant cell wall (CW) is a physical barrier that plays a dual role in plant physiology, providing structural support for growth and development. Understanding the dynamics of CW growth is crucial for optimizing crop yields. In this study, we employed onion (Allium cepa L.) epidermis as a model system, leveraging its layered organization to investigate growth stages. Microscopic analysis revealed proportional variations in cell size in different epidermal layers, offering insights into growth dynamics and CW structural adaptations. Fourier transform infrared spectroscopy (FTIR) identified 11 distinct spectral intervals associated with CW components, highlighting structural modifications that influence wall elasticity and rigidity. Biochemical assays across developmental layers demonstrated variations in cellulose, soluble sugars, and antioxidant content, reflecting biochemical shifts during growth. The differential expression of ten cell wall enzyme (CWE) genes, analyzed via RT-qPCR, revealed significant correlations between gene expression patterns and CW composition changes across developmental layers. Notably, the gene expression levels of the pectin methylesterase and fucosidase enzymes were associated with the contents in cellulose, soluble sugar, and antioxidants. To complement these findings, machine learning models, including Support Vector Machines (SVM), k-Nearest Neighbors (kNN), and Neural Networks, were employed to integrate FTIR data, biochemical parameters, and CWE gene expression profiles. Our models achieved high accuracy in predicting growth stages. This underscores the intricate interplay among CW composition, CW enzymatic activity, and growth dynamics, providing a predictive framework with applications in enhancing crop productivity and sustainability. Full article

The bacterial flagellar motor is one of the few known rotary motors, powering motility and chemotaxis. The mechanisms underlying its rotation and the switching of its rotational direction are fundamental problems in biology that are of significant interest. Recent high-resolution studies of the flagellar motor have transformed our understanding of the motor, revealing a novel gear mechanism where a membranous pentamer of MotA proteins rotates around a cell wall-anchored dimer of MotB proteins to turn the contacting flagellar rotor. A derivative model suggests that significant changes in rotor diameter occur during switching, enabling each MotA₅MotB₂ stator unit to shift between internal and external gear configurations, causing clockwise (CW) and counterclockwise (CCW) motor rotation, respectively. However, recent structural work favors a mechanism where the stator units dynamically swing back and forth between the two gear configurations without significant changes in rotor diameter. Given the intricate link between the switching model and the gear mechanism for flagellar rotation, a critical evaluation of the underlying assumptions is crucial for refining switching models. This review scrutinizes key assumptions within prevailing models of flagellar rotation and switching, identifies knowledge gaps, and proposes avenues for future biophysical tests. Full article

Osmotic stress impacts the cell wall properties in plants. This study aimed to elucidate the mechanisms involved in cell wall remodeling in etiolated (dark-grown) rice (Oryza sativa L.) shoots grown under polyethylene glycol (PEG)-induced osmotic stress conditions. Shoot growth was inhibited by 70% by the treatment with 60 mM PEG for 2 days. However, when the stressed seedlings were transferred to a solution without PEG, their shoot growth rate increased significantly. A measurement of the cell wall mechanical properties revealed that the cell walls of the stressed shoots became looser and more extensible than those of unstressed shoots. Among the cell wall constituents, the amounts of cell wall-bound phenolic acids, such as ferulic acid (FA), p-coumaric acid (p-CA), and diferulic acid (DFA), per shoot and per unit of matrix polysaccharide content were significantly reduced in the stressed shoots compared to those in the unstressed shoots. Concerning the formation of cell wall-bound phenolic acids, the activity of cell wall-bound peroxidase (CW-PRX) per unit of cell wall content, which is responsible for the coupling reaction of FA to produce DFA, was 3.5 times higher in stressed shoots than in unstressed shoots, while the activity was reduced by 20% on a shoot basis in stressed shoots compared to that in unstressed shoots. The expression levels of the major class III peroxidase genes in stressed shoots were either comparable to or slightly lower than those in unstressed shoots. Conversely, the phenylalanine ammonia-lyase (PAL) activity, which contributes to the biosynthesis of FA and p-CA, was reduced by 55% and 30% on a shoot and unit-of-protein-content basis, respectively, in stressed shoots compared to that in unstressed shoots. The expression levels of abundantly expressed PAL genes decreased by 14–46% under osmotic stress. Moreover, the gene expression levels of specific BAHD acyltransferases, which are responsible for the addition of FA and p-CA to form ester-linked moieties on cell wall constituents, decreased by 15–33% under osmotic stress. These results suggest that the downregulation of the expression of specific PAL and BAHD acyltransferase genes in osmotically stressed rice shoots is responsible for a reduction in the formation of cell wall-bound phenolic acid monomers. This, in turn, may result in a decrease in the levels of DFAs. The reduction in the formation of DFA-mediated cross-linking structures within the cell wall may contribute to an increase in the mechanical extensibility of the cell wall. The remodeling of cell walls in an extensible and loosened state could assist in maintaining the growth capacity of etiolated rice shoots grown under osmotic stress and contribute to rapid growth recovery following the alleviation of osmotic stress. Full article

Bgl2p is a major, conservative, constitutive glucanosyltransglycosylase of the yeast cell wall (CW) with amyloid amino acid sequences, strongly non-covalently anchored in CW, but is able to leave it. In the environment, Bgl2p can form fibrils and/or participate in biofilm formation. Despite a long study, the question of how Bgl2p is anchored in CW remains unclear. Earlier, it was demonstrated that Bgl2p lost the ability to attach in CW and to fibrillate after the deletion of nine amino acids in its C-terminal region (CTR). Here, we demonstrated that a Bgl2p anchoring is weakened by substitution Glu-233/Ala in the active center. Using AlphaFold and molecular modeling approach, we demonstrated the role of CTR on Bgl2p attachment and supposed the conformational possibilities determined by the presence or absence of an intramolecular disulfide bond, forming by Cys-310, leading to accessibility of amyloid sequence and β-turns localized in CTR of Bgl2p for protein interactions. We hypothesized the mode of Bgl2p attachment in CW. Using atomic force microscopy, we investigated fibrillar structures formed by peptide V187MANAFSYWQ196 and suggested that it can serve as a factor leading to the induction of amyloid formation during interaction of Bgl2p with other proteins and is of medical interest being located close to the surface of the molecule. Full article

In conifers, compression wood (CW) with a high lignin content forms at the base of the stem or branch in response to gravity, which is a good model system for studying lignin-rich wood formation. In this study, we identified and characterized the laccase gene family (PdeLAC) in Korean red pine (Pinus densiflora), which is integral to monolignol polymerization. Phylogenetic analysis of 54 PdeLAC genes with those from gymnosperms (i.e., Pinus taeda and Picea abies) and angiosperms (i.e., Populus trichocarpa, Arabidopsis thaliana, and Oryza sativa) revealed their categorization into five groups, highlighting distinct evolutionary relationships compared to angiosperms. Gene structure and motif analysis showed conserved copper-binding loops and variable substrate-binding loops, suggesting functional diversity. Expression profiling indicated that 23 PdeLAC genes, including three (PdeLAC28, PdeLAC1, and PdeLAC31) homologous to AtLAC17, were upregulated in developing xylem during the growing season, particularly in CW. Transgenic poplars overexpressing PdeLAC28 exhibited increased xylem area, cell wall thickness, and Klason lignin content, underscoring its role in lignin biosynthesis and CW formation. This study provides valuable insights into the molecular regulation of lignin biosynthesis in CW of P. densiflora, setting a foundation for advancing our understanding of wood formation mechanisms in gymnosperms. Full article

Percutaneous Coronary Intervention (PCI) is a treatment method that involves reopening narrowed arteries with a balloon catheter that delivers a cylindrical, mesh-shaped implant device to the site of the stenosis. Currently, by applying a coating to a bare metal stent (BMS) surface to improve biocompatibility, the main risks after PCI, such as restenosis and thrombosis, are reduced while maintaining the basic requirements for the mechanical behavior of the stent itself. In this work, for the first time, the development and optimization process of the spatial structure of the Co-Cr stent (L-605) with a graphene-based coating using cold-wall chemical vapor deposition (CW-CVD) to ensure uniform coverage of the implant was attempted. The CW-CVD process allows the coating of 3D structures, minimizing thermal stress on the surrounding equipment and allowing the deposition of coatings on temperature-sensitive materials. It produces uniform and high-purity films with control over the thickness and composition. The reduced heating of the chamber walls minimizes unwanted reactions, leading to fewer impurities in the final coating. The graphene layers obtained using Raman spectroscopy at different parameters of the CW-CVD process were verified, their properties were investigated, and the functional mechanical behavior of the studied graphene-covered stent was confirmed. In vitro, graphene-coated stents promoted rapid endothelial cell repopulation, an advantage over gold-standard drug-eluting stents delaying re-endothelialization. Also, full-range biocompatibility studies on potential allergic, irritation, toxicological, and pyrogenic reactions of new material in vivo on small animal models demonstrated excellent biocompatibility of the graphene-coated stents. Full article

Nitric oxide (NO) is an important signaling molecule involved in regulating plant processes to cope with abiotic stress. S-allyl-L-cysteine (SAC) is known to induce NO synthesis in animals. However, it is unknown whether SAC can trigger NO biosynthesis, regulate Cd transport, or alleviate Cd stress in plants. After being sprayed with 0.2 mM SAC, rice seedlings had a NO content that was 1.8 times higher than that of the control (ctrl) group at the ninth hour, which then gradually decreased. The expressions of Cd uptake and transport genes in the roots (including OsNRAMP5, OsNRAMP1, and OsHMA2) were markedly downregulated by 27.2%, 24.8%, and 49.1%, respectively, 72 h after SAC spraying treatment. The Cd content in seedling roots’ cell wall (CW) components significantly increased by 43.5% compared to that of the ctrl group. The Cd content in the shoots and roots decreased by 49.0% and 29.8%, respectively. Cd stress in the seedlings was also substantially alleviated. In conclusion, spraying rice seedlings with SAC triggered an increase in NO synthesis, regulated the expression of genes related to Cd transport, increased Cd fixation in the root CW components, and reduced Cd accumulation in the roots and shoots. Full article

Limited data are available on copper (Cu)–pH interaction-responsive genes and/or metabolites in plant roots. Citrus sinensis seedlings were treated with 300 μM (Cu toxicity) or 0.5 μM (control) CuCl₂ at pH 3.0 or 4.8 for 17 weeks. Thereafter, gene expression and metabolite profiles were obtained using RNA-Seq and widely targeted metabolome, respectively. Additionally, several related physiological parameters were measured in roots. The results indicated that elevating the pH decreased the toxic effects of Cu on the abundances of secondary metabolites and primary metabolites in roots. This difference was related to the following several factors: (a) elevating the pH increased the capacity of Cu-toxic roots to maintain Cu homeostasis by reducing Cu uptake and Cu translocation to young leaves; (b) elevating the pH alleviated Cu toxicity-triggered oxidative damage by decreasing reactive oxygen species (ROS) formation and free fatty acid abundances and increasing the ability to detoxify ROS and maintain cell redox homeostasis in roots; and (c) increasing the pH prevented root senescence and cell wall (CW) metabolism impairments caused by Cu toxicity by lowering Cu levels in roots and root CWs, thus improving root growth. There were some differences and similarities in Cu–pH interaction-responsive genes and metabolites between leaves and roots. Full article

Expansins are cell wall (CW) proteins that mediate the CW loosening and regulate salt tolerance in a positive or negative way. However, the role of Populus trichocarpa expansin A6 (PtEXPA6) in salt tolerance and the relevance to cell wall loosening is still unclear in poplars. PtEXPA6 gene was transferred into the hybrid species, Populus alba × P. tremula var. glandulosa (84K) and Populus tremula × P. alba INRA ‘717-1B4’ (717-1B4). Under salt stress, the stem growth, gas exchange, chlorophyll fluorescence, activity and transcription of antioxidant enzymes, Na⁺ content, and Na⁺ flux of root xylem and petiole vascular bundle were investigated in wild-type and transgenic poplars. The correlation analysis and principal component analysis (PCA) were used to analyze the correlations among the characteristics and principal components. Our results show that the transcription of PtEXPA6 was downregulated upon a prolonged duration of salt stress (48 h) after a transient increase induced by NaCl (100 mM). The PtEXPA6-transgenic poplars of 84K and 717-1B4 showed a greater reduction (42–65%) in stem height and diameter growth after 15 days of NaCl treatment compared with wild-type (WT) poplars (11–41%). The Na⁺ accumulation in roots, stems, and leaves was 14–83% higher in the transgenic lines than in the WT. The Na⁺ buildup in the transgenic poplars affects photosynthesis; the activity of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT); and the transcription of PODa2, SOD [Cu-Zn], and CAT1. Transient flux kinetics showed that the Na⁺ efflux of root xylem and leaf petiole vascular bundle were 1.9–3.5-fold greater in the PtEXPA6-transgenic poplars than in the WT poplars. PtEXPA6 overexpression increased root contractility and extensibility by 33% and 32%, indicating that PtEXPA6 increased the CW loosening in the transgenic poplars of 84K and 717-1B4. Noteworthily, the PtEXPA6-promoted CW loosening was shown to facilitate Na⁺ efflux of root xylem and petiole vascular bundle in the transgenic poplars. We conclude that the overexpression of PtEXPA6 leads to CW loosening that facilitates the radial translocation of Na⁺ into the root xylem and the subsequent Na⁺ translocation from roots to leaves, resulting in an excessive Na⁺ accumulation and consequently, reducing salt tolerance in transgenic poplars. Therefore, the downregulation of PtEXPA6 in NaCl-treated Populus trichocarpa favors the maintenance of ionic and reactive oxygen species (ROS) homeostasis under long-term salt stress. Full article

This review covers a group of non-covalently associated molecules, particularly proteins (NCAp), incorporated in the yeast cell wall (CW) with neither disulfide bridges with proteins covalently attached to polysaccharides nor other covalent bonds. Most NCAp, particularly Bgl2, are polysaccharide-remodeling enzymes. Either directly contacting their substrate or appearing as CW lipid-associated molecules, such as in vesicles, they represent the most movable enzymes and may play a central role in CW biogenesis. The absence of the covalent anchoring of NCAp allows them to be there where and when it is necessary. Another group of non-covalently attached to CW molecules are polyphosphates (polyP), the universal regulators of the activity of many enzymes. These anionic polymers are able to form complexes with metal ions and increase the diversity of non-covalent interactions through charged functional groups with both proteins and polysaccharides. The mechanism of regulation of polysaccharide-remodeling enzyme activity in the CW is unknown. We hypothesize that polyP content in the CW is regulated by another NCAp of the CW—acid phosphatase—which, along with post-translational modifications, may thus affect the activity, conformation and compartmentalization of Bgl2 and, possibly, some other polysaccharide-remodeling enzymes. Full article

In nature, arbuscular mycorrhizal fungi (AMF) play a crucial role in the root systems of plants. They can help enhance the resistance of host plants by improving the compartmentalization of toxic metal contaminants in the cell walls (CWs). However, the functions and responses of various CW subfractions to mycorrhizal colonization under Cd exposure remain unknown. Here we conducted a study to investigate how Cd is stored in the cell walls of maize roots colonized by Funneliformis mosseae. Our findings indicate that inoculating the roots with AMF significantly lowers the amount of Cd in the maize shoots (63.6 ± 6.54 mg kg⁻¹ vs. 45.3 ± 2.19 mg kg⁻¹, p < 0.05) by retaining more Cd in the mycorrhized roots (224.0 ± 17.13 mg kg⁻¹ vs. 289.5 ± 8.75 mg kg⁻¹, p < 0.01). This reduces the adverse effects of excessive Cd on the maize plant. Additional research on the subcellular distribution of Cd showed that AMF colonization significantly improves the compartmentalization of 88.2% of Cd in the cell walls of maize roots, compared to the 80.8% of Cd associated with cell walls in the non-mycorrhizal controls. We observed that the presence of AMF did not increase the amount of Cd in pectin, a primary binding site for cell walls; however, it significantly enhanced the content of lignin and the proportion of Cd in the total root cell walls. This finding is consistent with the increased activity of lignin-related enzymes, such as PAL, 4CL, and laccase, which were also positively impacted by mycorrhizal colonization. Fourier transform infrared spectroscopy (FTIR) results revealed that AMF increased the number and types of functional groups, including −OH/−NH and carboxylate, which chelate Cd in the lignin. Our research shows that AMF can improve the ability of maize plants to tolerate Cd by reducing the amount of Cd transferred from the roots to the shoots. This is achieved by increasing the amount of lignin in the cell walls, which binds with Cd and prevents it from moving through the plant. This is accomplished by activating enzymes related to lignin synthesis and increasing the exposure of Cd-binding functional groups of lignin. However, more direct evidence on the immobilization of Cd in the mycorrhiza-altered cell wall subfractions is needed. Full article