Wall Components Research Articles

The Bacterial Cell Wall Components Lipopolysaccharide and Peptidoglycan Initiate Divergent Local Tissue and Systemic Inflammatory Response Profiles in the Chicken Model

The innate immune system plays an important role in the defense against pathogens, whereby the ability to rapidly mount an effective inflammatory response is critical in the elimination/containment of the infection. To better understand the nature of the inflammatory responses to bacterial components in chickens, we used the growing feather (GF) cutaneous bioassay together with blood sampling to examine the local and systemic inflammatory responses initiated by intradermal (i.d.) GF-pulp injection of lipopolysaccharide (LPS) from Salmonella Typhimurium or peptidoglycan (PGN) from Staphylococcus aureus. Three studies were conducted in egg-type chickens between 9 and 15 weeks of age; Study 1 and 2 examined the leukocyte response profiles to a 100-fold dose range of LPS or PGN over 24 h or 7 d, respectively; Study 3 examined the leukocyte- and cytokine mRNA-profiles in pulps in response to LPS and PGN concurrently over 72 h. I.d. injection of LPS stimulated a heterophil and monocyte/macrophage dominated response in both GF-pulps and blood that was resolved by 48–72 h and differed based on dose administered. The inflammatory response stimulated by PGN was characterized by rapid infiltration of lymphocytes in GF-pulps with sustained high levels of T and B cells over 5–7 d and was neither affected by PGN dose nor reflected in the blood. Limited cytokine transcriptome analyses did not reveal differences that could explain the divergent response profiles to LPS versus PGN. More research is needed to understand the mechanisms underlying the divergent inflammatory responses to LPS and PGN in chickens.

Immunomodulatory Properties of Multi-Strain Postbiotics on Human CD14+ Monocytes

The ability of probiotics, comprising live microbiota, to modulate the composition of intestinal microbiomes has been connected to modulation of the central nervous system (Gut–Brain axis), neuroendocrine system (Gut–Skin axis), and immune response (Gut–Immune axis). Less information is known regarding the ability of postbiotics (cell wall components and secreted metabolites derived from live organisms) to regulate host immunity. In the present study, we tested postbiotics comprising single strains of bacteria and yeast (Lactobacillus acidophilus 16axg, Lacticaseibacillus rhamnosus 18fx, Saccharomyces cerevisiae var. boulardii 16mxg) as well as combinations of multiple strains for their ability to stimulate cytokine production by human CD14+ monocytes. We quantified cytokine gene and protein expression levels in monocytes following stimulation with postbiotics. Both heat-killed L. acidophilus and L. rhamnosus stimulated naïve monocytes without significant differences between them. Heat-killed S. boulardii stimulated less cytokine production compared to postbiotic bacteria at the same concentration. Interestingly, the addition of heat-killed yeast to heat-killed L. acidophilus and L. rhamnosus resulted in an enhancement of immune stimulation. Thus, heat-killed postbiotics have immune-modulating potential, particularly when bacteria and yeast are combined. This approach may hold promise for developing targeted interventions that can be fine-tuned to modulate host immune response with beneficial health impact.

Paralogous Gene Recruitment in Multiple Families Constitutes Genetic Architecture and Robustness of Pod Dehiscence in Legumes.

Pod dehiscence facilitates seed dispersal in wild legumes while indehiscence is a key domestication trait in cultivated ones. However, the evolutionary genetic mechanisms underlying its diversity are largely unclear. In this study, we compared transcriptomes of two warm-season (Glycine spp. and Phaseolus spp.) and two cool-season (Pisum spp. and Medicago ruthenica) legumes in analysis of dehiscent and indehiscent pod genotypes. Differentially expressed genes in AP2/ERF-like transcription factors and seven structural gene families, including lactoperoxidase, laccase, and cellulose synthase-interactive proteins, which are involved in secondary cell wall component accumulation, were identified to exert key roles in pod dehiscence variation. In accordance with this, higher lignin and cellulose contents were observed in pod secondary cell wall of dehiscent accessions of soybean and pea; however, the variation patterns of lignin polymers in soybean (accumulation) and pea (proportion) differed between dehiscent and indehiscent pods. Moreover, genome-wide comparative analysis revealed that orthogroups represented <1% of all identified differentially expressed genes could be traced among the four genera of legumes, while recruiting paralogous members may constitute the genetic robustness of legume pod dehiscence. This study compared the genetic mechanism among several legumes in pod dehiscence formation and revealed a compensating role of paralogous redundancy of involved gene families in seed dispersal, which can guide crop breeding.

Foliar uniconazole application increases rice lodging resistance by altering stem morphological and anatomical traits

Lodging is a limiting factor for rice production in the Sichuan Basin, China. However, the mechanisms of stem lodging resistance, especially its regulation by plant growth regulators are still unclear. A two-year field study, by using the three foliar application rates of uniconazole with two rice varieties, Yuxiang203 (YX203) and C-Liangyouhuazhan (CLYHZ), was conducted to determine stem lodging resistance and its morphological and anatomical mechanisms in rice plants. The results revealed that, compared with that in 2019, the grain yield in 2020 significantly decreased, whereas the lodging index (LI) significantly increased. Uniconazole treatment increased the rice yield by 4.6%−11.2% and 2.1%−7.0%, and decreased LI by 21.1%−33.9% and 11.4%−29.6% in YX203 and CLYHZ, respectively. Uniconazole treatment shortened the length of the basal internodes by 19.5%−33.0% (YX203) and 24.7%−40.7% (CLYHZ), resulting in a significant reduction in plant height. Uniconazole treatment increased the mechanical tissue thickness, areas of small and larger vascular bundles, and culm diameter and further increased the breaking strength of the two varieties. Cell wall components, including cellulose and lignin, were increased by foliar application of uniconazole, thereby creating denser sclerenchyma cells and increasing the thickness of the mechanical tissue and area of the vascular bundle. These results suggest that the application of uniconazole enhances stem mechanical strength via increased mechanical tissue thickness and larger areas of small and large vascular bundles, thereby improving the lodging resistance of rice plants.

Application of BIM in Modeling Concrete Retaining Walls

During the hydraulic design phase, challenges often arise, such as poor coordination between different disciplines and ineffective information transfer. These issues can lead to errors in the design process and unreasonable structural designs. As the complexity of engineering projects continues to increase, traditional 2D design methods can no longer meet current demands. As a result, Building Information Modeling (BIM) technology has been introduced, shifting the design approach to 3D modeling. This transition has significantly improved design accuracy and construction efficiency. To further optimize this process, a user interface interaction system based on C# language and the Revit API was developed. Through this system, we successfully implemented the 3D automatic modeling of retaining wall components and a reinforcement plugin. These tools have greatly saved time on repetitive tasks for designers, improving work efficiency. At the same time, they have advanced the informatization of hydraulic concrete structure construction, making the construction process more efficient, precise, and controllable. The 3D deepening design can accurately express the geometric characteristics of the structure, enhancing precision and accelerating the informatization development of hydraulic concrete structure construction.

Engineering the secretome of Aspergillus niger for cellooligosaccharides production from plant biomass

BackgroundFermentation of sugars derived from plant biomass feedstock is crucial for sustainability. Hence, utilizing customized enzymatic cocktails to obtain oligosaccharides instead of monomers is an alternative fermentation strategy to produce prebiotics, cosmetics, and biofuels. This study developed an engineered strain of Aspergillus niger producing a tailored cellulolytic cocktail capable of partially degrading sugarcane straw to yield cellooligosaccharides.ResultsThe A. niger prtT∆ strain created resulted in a reduced extracellular protease production. The prtT∆ background was then used to create strains by deleting exoenzyme encoding genes involved in mono- or disaccharide formation. Consequently, we successfully generated a tailored prtT∆bglA∆ strain by eliminating a beta-glucosidase (bglA) gene and subsequently deleted two cellobiohydrolases and one beta-xylosidase encoding genes using a multiplex strategy, resulting in the Quintuple∆ strain (prtT∆; bglA∆; cbhA∆; cbhB∆; xlnD∆). When applied for sugarcane biomass degradation, the tailored secretomes produced by A. niger resulted in a higher ratio of cellobiose and cellotriose compared with glucose relative to the reference strain. Mass spectrometry revealed that the Quintuple∆ strain secreted alternative cellobiohydrolases and beta-glucosidases to compensate for the absence of major cellulases. Enzymes targeting minor polysaccharides in plant biomass were also upregulated in this tailored strain.ConclusionTailored secretome use increased COS/glucose ratio during sugarcane biomass degradation showing that deleting some enzymatic components is an effective approach for producing customized enzymatic cocktails. Our findings highlight the plasticity of fungal genomes as enzymes that target minor components of plant cell walls, and alternative cellulases were produced by the mutant strain. Despite deletion of important secretome components, fungal growth was maintained in plant biomass.

Effect of UV-B stress on olive (Olea europaea L.) pollen tubes: A study of callose plug deposition and male germ unit integrity.

While UV-B radiation is beneficial to plant growth, it can also cause adverse effects. The pollen tube, a key component of plant reproduction with a tip growth mechanism, is an excellent cellular model for understanding how environmental stressors such as UV-B radiation affect plant cell growth. This research investigated the effect of UV-B on olive pollen both before and after germination. Pollen grains were hydrated and exposed to UV-B radiation for 1h. Pollen tube germination was then evaluated 4 and 24h after exposure. To study the effect of UV-B radiation on developing pollen tubes, pollen was germinated for 4h prior to 1h of UV-B exposure. Pollen tube growth was evaluated by assessing the distribution of cell wall components, the distance between callose plugs and nuclei, and the distance between the male germ unit and the pollen tube tip. We also examined the accumulation of callose synthase. The results showed that UV-B radiation significantly inhibited the growth of pollen tubes, thereby preventing successful fertilization. The effect of UV-B exposure on pollen tube growth was mainly due to the alteration of position of callose plugs and the level of callose synthase in the pollen tube, potentially affecting its growth. In addition, UV-B radiation affected the movement and integrity of the male germ unit, a critical element for successful fertilization. This research sheds light on how UV-B radiation affects the growth of pollen tubes and highlights the need for further research into the effects of UV-B radiation on plant cells and plant reproduction.

Post-mortem analysis of material deposition and fuel retention on the plasma-facing materials after the 2021 campaign in EAST

Plasma-wall interaction (PWI) is a critical concern in tokamaks because of its significant impact on the lifetimes of plasma-facing materials (PFMs), fuel retention, and plasma performance. In 2020, the Experimental Advanced Superconducting Tokamak (EAST) PFMs were upgraded to predominantly metallic walls. Following the 2021 experimental campaign, the deposition distribution and fuel retention on the surfaces of the PFMs along both the poloidal and toroidal directions were analyzed. The poloidal tests commenced at the high-field side (HFS), proceeded to the lower divertor, then to the low-field side (LFS), and finally to the upper divertor. The toroidal tests were performed at the midplane of the HFS, beginning from port A and ending at port P. The distributions of the deposits in the poloidal and toroidal directions were clearly asymmetrical. Mo and Fe particles sputtered from the first wall and inner stainless steel (SS) components were prone to deposition in the lower divertor region, as evidenced by the fact that the elemental content in the far-SOL region of the inner divertor exhibited Mo and Fe peaks, as did both the near- and far-SOL regions of the outer divertor. In addition, quick re-deposition of W and Fe was observed, as demonstrated by the fact that their contents near the erosion sources were higher than those farther away along the toroidal first wall on the HFS. This tendency was stronger for W than for Fe. Further analysis indicated that the deposits consisted of Li, C, O, W, Mo, Fe, Cu, and Ni. The Li and Fe contents were much higher than those of other metal impurities, with peak values of 8.17 μg/mm2 and 7.78 μg/mm2, respectively. The Li content decreased along the HFS first wall from 8.17 μg/mm2 at P-2 to 1.82 μg/mm2 at P-22, and then increased to 2.72 μg/mm2 at P-32 in the divertor, while the Fe content was higher around the top side and the midplane of the HFS. The deposited Li originated from routine wall conditioning and existed primarily in the form of Li2CO3. Additionally, other metal impurities deposited on the Li2CO3 surfaces exhibited various irregular shapes, often appearing as aggregated and recrystallized small particles. Furthermore, significant deuterium (D) retention on the order of 1021 atoms/m2 was measured on all the analyzed SiC-coated graphite tiles on the HFS. The D content at location P-10 at the midplane was the highest along the poloidal direction of the HFS because it was directly facing the NBI beam.

Dynamic changes of heterogeneous cell wall macromolecules in differentiating conifer xylem using cytochemical localization

Tracing dynamic changes of heterogeneous cell wall components during xylem differentiation is essential for understanding the intricate architecture of wood cell walls at the individual secondary cell wall layer level. Here we employ histochemical- and immunological approaches to visualize the deposition of cellular polymers during xylem differentiation in Pinus bungeana. In axial tracheids, deposition of crystalline cellulose and glucomannan preceded xylan and lignin. Lignification was initiated in primary cell wall corners during development of the S1 layer and intensified with cell wall thickening. Immunofluorescence labeling showed an earlier deposition of glucomannan than xylan with strong presence in S1 layer corner regions at early stages of differentiation. Quantification of immunogold-labeled xylan and glucomannan showed distinct increasing trends during thickening of tracheid wall layers with xylan labeling of the S1 and S2 layers at the S3 stage greater than the S2 stage. Differential cell wall polymer deposition was evident in mature tracheid areas with glucomannan absent in warty layers. Pectins were highly concentrated in unlignified primary cell walls but decreased with axial tracheid wall differentiation. The sequence of polymer deposition in ray cells was similar but lagged behind axial tracheids with ray parenchyma remaining unlignified with thinner cell walls than ray tracheids.

AtFH5 recruits and transports the arabinogalactan protein AGP23 to maintain the tip growth of pollen tube

Actin cytoskeleton drives the targeted transport of cell wall components to sustain the tip growth of pollen tubes for double fertilization; however, the underlying mechanism remains largely unknown. Arabidopsis formin 5 (AtFH5), an actin-nucleating protein, localizes at secretory vesicles and mediates actin polymerization-based vesicle trafficking in pollen. Here, we demonstrate that AtFH5 determines the recruitment and transport of cell wall components in AtFH5-labeled vesicles during the tip growth of pollen tubes. Through a screen of interacting proteins of AtFH5, we identify many cell wall-related proteins, with arabinogalactan protein 23 (AGP23) occupying the highest frequency. AtFH5 interacts with AGP23 via its N-terminal extracellular domain (ECD) and jointly regulate the pollen germination and tube growth process. Further observations reveal that AGP23 co-localizes with AtFH5 at moving vesicles, germination sites, and pollen tube tips, suggesting that AGP23 is delivered by AtFH5-labeled vesicles. Deletion of the ECD of AtFH5 interrupts the dynamic localization and cell-wall connection of AGP23 in pollen grains and tubes. Cytological and genetic evidence shows that AGP23 and AtFH5 work in the same pathway to modulate cell wall composition. Together, our data uncover a role of formin in directing the sorting and deposition of cell wall components via secretory vesicle trafficking during pollen germination and tube growth.

Preliminary exploration of the mechanisms underlying morphology engineering of Monascus purpureus during submerged fermentation via multi-omics approaches

Herein, multi-omics approaches were employed to investigate the relationship between mycelia morphology and biosynthesis of Monascus pigments (MPs) during submerged fermentation of Monascus purpureus. Through heavy ion beam irradiation, eleven mutants with diverse mycelial morphologies and MPs productions were generated. Genome resequencing analysis revealed that the autologous genes of these mutants were primarily enriched in pathways associated with histone ubiquitination and methylation. Among them, M. purpureus ZS exhibited a 2.49-fold increase in MPs production and distinct mycelia morphology compared to the parent strain, thus it was selected for transcriptomic and metabolomic analyses. The results revealed showed significant alterations in lysine metabolism, lipid metabolism, as well as cell wall and membrane components. Additionally, upregulated CoA biosynthesis along with type I polyketide structures and the genetic cluster responsible for MPs biosynthesis facilitated accumulation of MPs. These findings provide a theoretical foundation for morphological engineering of Monascus while offering novel strategies to achieve higher levels of MPs production during submerged fermentation.

Bio-organic fertilizer affects secondary cell wall biosynthesis of Dendrocalamus farinosus by inhibiting the phenylpropanoid metabolic pathway

Bamboo, as a timber plant, holds significant environmental and economic value. Dendrocalamus farinosus is particularly valuable as it serves both as a source of bamboo shoots and timber, offering high yield, strong disease resistance, and superior fiber quality. Our previous study demonstrated that bio-organic fertilizers promoted the growth of D. farinosus and significantly altered the cellulose and lignin content, key components of the secondary cell wall in culms. However, the underlying regulatory mechanisms remain unclear. In this study, we used metabolomic and transcriptomic analyses to uncover the potential mechanisms by which bio-organic fertilizers affect the secondary cell wall biosynthesis in D. farinosus. A total of 1,437 metabolites were identified, with 20 differential metabolites significantly enriched in the phenylpropanoid metabolic pathway in bamboo shoots (7 upregulated; 13 downregulated). We identified 8,075 differentially expressed genes in bamboo shoots, including 72 genes potentially involved in lignin and flavonoid biosynthesis (6 upregulated; 66 downregulated). In internodes, we identified 5,324 differentially expressed genes, including 83 genes potentially involved in secondary cell wall biosynthesis (43 upregulated; 39 downregulated). Quantitative real-time PCR (qRT-PCR) validated the expression patterns of 8 key genes in internodes. The results suggest that bio-organic fertilizers may affect secondary cell wall biosynthesis in internodes by inhibiting the phenylpropanoid metabolic pathway in D. farinosus shoots. Our study offers insights into the efficient utilization of bamboo and lignocellulosic biomass, serving as a valuable resource for future research.

Deciphering the Genetic and Biochemical Drivers of Fruit Cracking in Akebia trifoliata.

This study investigates the molecular mechanisms underlying fruit cracking in Akebia trifoliata, a phenomenon that significantly impacts fruit quality and marketability. Through comprehensive physiological, biochemical, and transcriptomic analyses, we identified key changes in cell wall components and enzymatic activities during fruit ripening. Our results revealed that ventral suture tissues exhibit significantly elevated activities of polygalacturonase (PG) and β-galactosidase compared to dorsoventral line tissues, indicating their crucial roles in cell wall degradation and structural weakening. The cellulose content in VS tissues peaked early and declined during ripening, while DL tissues maintained relatively stable cellulose levels, highlighting the importance of cellulose dynamics in fruit cracking susceptibility. Transcriptomic analysis revealed differentially expressed genes (DEGs) associated with pectin biosynthesis and catabolism, cell wall organization, and oxidoreductase activities, indicating significant transcriptional regulation. Key genes like AKT032945 (pectinesterase) and AKT045678 (polygalacturonase) were identified as crucial for cell wall loosening and pericarp dehiscence. Additionally, expansin-related genes AKT017642, AKT017643, and AKT021517 were expressed during critical stages, promoting cell wall loosening. Genes involved in auxin-activated signaling and oxidoreductase activities, such as AKT022903 (auxin response factor) and AKT054321 (peroxidase), were also differentially expressed, suggesting roles in regulating cell wall rigidity. Moreover, weighted gene co-expression network analysis (WGCNA) identified key gene modules correlated with traits like pectin lyase activity and soluble pectin content, pinpointing potential targets for genetic manipulation. Our findings offer valuable insights into the molecular basis of fruit cracking in A. trifoliata, laying a foundation for breeding programs aimed at developing crack-resistant varieties to enhance fruit quality and commercial viability.

Sulfonamide-Based Inhibition of the β-Carbonic Anhydrase from A. baumannii, a Multidrug-Resistant Bacterium.

Acinetobacter baumannii is a Gram-negative opportunistic pathogen responsible for severe hospital-associated infections. Owing to its ability to develop resistance to a wide range of antibiotics, novel therapeutic strategies are urgently needed. One promising approach is to target bacterial carbonic anhydrases (CAs; EC 4.2.1.1), which are enzymes critical for various metabolic processes. The genome of A. baumannii encodes a β-CA (βAbauCA), which is essential for producing bicarbonate ions required in the early stages of uridine triphosphate (UTP) synthesis, a precursor for the synthesis of peptidoglycans, which are vital components of the bacterial cell wall. This study aimed to inhibit βAbauCA in vitro, with the potential to impair the vitality of the pathogen in vivo. We conducted sequence and structural analyses of βAbauCA to explore its differences from those of human CAs. Additionally, kinetic and inhibition studies were performed to investigate the catalytic efficiency of βAbauCAβ and its interactions with sulfonamides and their bioisosteres, classical CA inhibitors. Our results showed that βAbauCA has a turnover rate higher than that of hCA I but lower than that of hCA II and displays distinct inhibition profiles compared to human α-CAs. Based on the obtained data, there are notable differences between the inhibition profiles of the human isoforms CA I and CA II and bacterial βAbauCA. This could open the door to designing inhibitors that selectively target bacterial β-CAs without affecting human α-CAs, as well as offer a novel strategy to weaken A. baumannii and other multidrug-resistant pathogens.

Examination of testicular veins in children with varicocele

BACKGROUND: Diagnostics and management of varicocele in children, adolescents and males are issues of annual discussions in the professional community, yet the debates still continue. Pathophysiological changes in the testicular veins in children with varicocele are dubious and require further in-depth fundamental researches. AIM: To study the difference in ultrastructural organization of testicular veins in boys with varicocele depending on the malformation degree. METHODS: In the present study, the authors examined the ultrastructure of 58 bioptats taken from spermatic veins during surgeries in adolescent boys aged 11–17 with varicocele of degree II and III. RESULTS: A comparative analysis of ultrastructural changes in the spermatic veins revealed that signs of endotheliocyte destruction of varying degree are met in 86% of biopsy samples in Group I and in 100% in Group II. Electron microscopy allowed to examine the muscular component of testicular veins in varicocele, as well as to examine the structure of connective tissue component in the venous wall, basing on an anatomical-functional paradigm of the connective tissue as the main "building brick" of organ and system structures. CONCLUSION: Ultrastructural changes in the testicular vein wall were revealed in children with varicocele, what indicates the presence of congenital malformation in this wall and of accompanied endothelial dysfunction. Foldings of the vascular wall with hollows and pockets, connective tissue loosening and separation of fibers, endothelium destruction and detachment were equally met in boys and adolescents with varicocele of degree II and III.

Lipopolysaccharide-mediated effects of the microbiota on sleep and body temperature

Recent research suggests that microbial molecules translocated from the intestinal lumen into the host’s internal environment may play a role in various physiological functions, including sleep. Previously, we identified that butyrate, a short-chain fatty acid produced by intestinal bacteria, and lipoteichoic acid, a cell wall component of gram-positive bacteria, induce sleep when their naturally occurring translocation is mimicked by direct delivery into the portal vein. Building upon these findings, we aimed to explore the sleep signaling potential of intraportally administered lipopolysaccharide (LPS), a primary component of gram-negative bacterial cell walls, in rats. Low dose of LPS (1 μg/kg) increased sleep duration and prolonged fever, without affecting systemic LPS levels. Interestingly, administering LPS systemically outside the portal region at a dose 20 times higher did not affect sleep, indicating a localized sensitivity within the hepatoportal region for the sleep and febrile effects of LPS. Furthermore, both the sleep- and fever-inducing effects of LPS were inhibited by indomethacin, a prostaglandin synthesis inhibitor, and replicated by intraportal administration of prostaglandin E2 or arachidonic acid, suggesting the involvement of the prostaglandin system in mediating these actions.

Evolution of g-type lysozymes in metazoa: insights into immunity and digestive adaptations.

Exploring the evolutionary dynamics of lysozymes is critical for advancing our knowledge of adaptations in immune and digestive systems. Here, we characterize the distribution of a unique class of lysozymes known as g-type, which hydrolyze key components of bacterial cell walls. Notably, ctenophores, and choanoflagellates (the sister group of Metazoa), lack g-type lysozymes. We reveal a mosaic distribution of these genes, particularly within lophotrochozoans/spiralians, suggesting the horizontal gene transfer events from predatory myxobacteria played a role in their acquisition, enabling specialized dietary and defensive adaptations. We further identify two major groups of g-type lysozymes based on their widespread distribution in gastropods. Despite their sequence diversity, these lysozymes maintain conserved structural integrity that is crucial for enzymatic activity, underscoring independent evolutionary pathways where g-type lysozymes have developed functionalities typically associated with different lysozyme types in other species. Specifically, using Aplysia californica as a reference species, we identified three distinct g-type lysozyme genes: two are expressed in organs linked to both feeding and defense, and the third exhibits broader distribution, likely associated with immune functions. These findings advance our understanding of the evolutionary dynamics shaping the recruitment and mosaic functional diversification of these enzymes across metazoans, offering new insights into ecological physiology and physiological evolution as emerging fields.

Molecular cloning and characterization of a brassinosteriod biosynthesis-related gene PtoCYP90D1 from Populus tomentosa

Brassinosteroids (BRs), one of the major classes of phytohormones are essential for various processes of plant growth, development, and adaptations to biotic and abiotic stresses. In Arabidopsis, AtCYP90D1 acts as a bifunctional cytochrome P450 monooxygenase, catalyzing C-23 hydroxylation in the brassinolide biosynthetic pathway. The present study reports the functional characterizations of PtoCYP90D1, one of the AtCYP90D1 homologous genes from Populus tomentosa. The qRT-PCR analysis showed that PtoCYP90D1 was highly expressed in roots and old leaves. Overexpression of PtoCYP90D1 (PtoCYP90D1-OE) in poplar promoted growth and biomass yield, as well as increased xylem area and cell layers. Transgenic plants exhibited a significant increase in plant height and stem diameter as compared to the wild type. In contrast, the CRISPR/Cas9-generated mutation of PtoCYP90D1 (PtoCYP90D1-KO) resulted in significantly decreased biomass production in transgenic plants. Further studies revealed that cell wall components increased significantly in PtoCYP90D1-OE lines but not in PtoCYP90D1-KO lines, as compared to wild-type plants. Overall, the findings indicate a positive role of PtoCYP90D1 in improving growth rate and elevating biomass production in poplar, which will have positive implications for its versatile industrial or agricultural applications.

Microbial communities and their role in enhancing hemp fiber quality through field retting.

The current development of industrial hemp "Cannabis Sativa L." fibers for technical textiles and industrial applications requires high-quality fibers with homogeneous properties. However, several factors have been reported to influence the fibers' intrinsic properties, including a post-harvest process known as retting. This process plays a crucial role in facilitating the mechanical extraction of fibers from hemp stems. Retting involves the degradation of the amorphous components surrounding the fiber bundles enabling their decohesion from stems. Microorganisms play a central role in mediating this bioprocess. During retting, they colonize the stems' surface. Therefore, the biochemical components of plant cell wall, acting as natural binding between fibers, undergo a breakdown through the production of microbial enzymes. Although its critical role, farmers often rely on empirical retting practices, and considering various biotic and abiotic factors, resulting in fibers with heterogenous properties. These factors limit the industrial applications of hemp fibers due to their inconsistent properties. Thus, the purpose of this review is to enhance our comprehension of how retting influences the dynamics of microbial communities and, consequently, the evolution of the biochemical properties of hemp stems throughout this process. Better understanding of retting is crucial for effective process management, leading to high-value fibers. KEY POINTS: • Retting enables degradation of cell wall components, controlling fiber properties. • Microbial enzymatic activity is crucial for successful decohesion of fiber bundles. • Understanding retting mechanisms is essential for consistent fiber production.

Cell wall nanoparticles from hyphae of Alternaria infectoria grown with caspofungin, nikkomycin, or pyroquilon trigger different activation profiles in macrophages.

Alternaria infectoria causes opportunistic human infections and is a source of allergens leading to respiratory allergies. In this work, we prepared cell wall nanoparticles (CWNPs) as a novel approach to study macrophage immunomodulation by fungal hyphal cell walls. A. infectoria was grown in the presence of caspofungin, an inhibitor of β(1,3)-glucan synthesis; nikkomycin Z, an inhibitor of chitin synthases; and pyroquilon, an inhibitor of dihydroxynaphthalene (DHN)-melanin synthesis. Distinct CWNPs were obtained from these cultures, referred to as casCWNPs, nkCWNPs, and pyrCWNPs, respectively. CWNPs are round-shaped particles with a diameter of 70-200 nm diameter particles that when added to macrophages are taken up by membrane ruffling. CWNPs with no DHN-melanin and more glucan (pyrCWNPs) caused early macrophage activation and lowest viability, with the cells exhibiting ultrastructural modifications such as higher vacuolization and formation of autophagy-like structures. CasCWNPs promoted the highest tumor necrosis factor alpha (TNF-α) and interleukin 1 beta (IL-1β) increase, also resulting in the release of partially degraded chitin, an aspect never observed in macrophage-like cells and fungi. After 6 h of interaction with CWNPs, only half were viable, except with control CWNPs. Overall, this work indicates that compounds that modify the fungal cell wall led to CWNPs with new properties that may have implications for the effects of drugs during antifungal therapy. CWNPs provide a new tool to study the interaction of hyphal fungal cell wall components with phagocytic cells and enable to show how the modification of cell wall components in A. infectoria can modulate the response by macrophages.IMPORTANCEAlternaria species are ubiquitous environmental fungi to which the human host can continuously be exposed, through the inhalation of fungal spores but also of fragments of hyphae, from desegregated mycelia. These fungi are involved in hypersensitization and severe respiratory allergies, such as asthma, and can cause opportunistic infections in immunodepressed human host leading to severe disease. The first fungal structures to interact with the host cells are the cell wall components, and their modulation leads to differential immune responses. Here, we show that fungal cells grown with cell wall inhibitors led to cell wall nanoparticles with new properties in their interaction with macrophages. With this strategy, we overcame the limitation of in vitro assays interacting with filamentous fungi and showed that the absence of DNH-melanin leads to higher virulence, while caspofungin leads to cells walls that trigger higher hydrolysis of chitin and higher production of cytokines.