Fiber Cells Research Articles

Anchorage of bacterial effector at plasma membrane via selective phosphatidic acid binding to modulate host cell signaling.

Binding phospholipid is a simple, yet flexible, strategy for anchorage of bacterial effectors at cell membrane to manipulate host signaling responses. Phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-biphosphate are the only two phospholipid species known to direct bacterial effectors to establish inner leaflet localization at the plasma membrane. Here, selectivity of phosphatidic acid (PA) by bacterial effectors for the plasma membrane anchorage and its molecular entity was identified. C-terminal BID domain of Bartonella T4SS effectors (Beps) directed the plasma membrane localization of Beps in host cells through binding with PA. A hydrophobic segment of the 'HOOK' subdomain from BID is inserted into the bilayer to enhance the interaction of positively charged residues with the lipid headgroups. Mutations of a conserved arginine facilitating the electrostatic interaction, a conserved glycine maintaining the stability of the PA binding groove, and hydrophobic residues determining membrane insertion, prevented the anchorage of Beps at the plasma membrane. Disassociation from plasma membrane to cytosol attenuated the BepC capacity to induce stress fiber formation and cell fragmentation in host cells. The substitution of alanine with aspartic acid at the -1 position preceding the conserved arginine residue hindered BepD anchoring at the plasma membrane, a vital prerequisite for its ability to elicit IL-10 secretion in host macrophages. In conclusion, our findings reveal the PA-binding properties of bacterial effectors to establish plasma membrane localization and will shed light on the intricate mechanisms employed by bacterial effectors within host cells.

Embryo movement is required for limb tendon maturation

IntroductionFollowing early cell specification and tenocyte differentiation at the sites of future tendons, very little is known about how tendon maturation into robust load-bearing tissue is regulated. Between embryonic day (E)16 and E18 in the chick, there is a rapid change in mechanical properties which is dependent on normal embryo movement. However, the tissue, cellular and molecular changes that contribute to this transition are not well defined.MethodsHere we profiled aspects of late tendon development (collagen fibre alignment, cell organisation and Yap pathway activity), describing changes that coincide with tissue maturation. We compared effects of rigid (constant static loading) and flaccid (no loading) immobilisation to gain insight into developmental steps influenced by mechanical cues.ResultsWe show that YAP signalling is active and responsive to movement in late tendon. Collagen fibre alignment increased over time and under static loading. Cells organise into end-to-end stacked columns with increased distance between adjacent columns, where collagen fibres are deposited; this organisation was lost following both types of immobilisation.DiscussionWe conclude that specific aspects of tendon maturation require controlled levels of dynamic muscle-generated stimulation. Such a developmental approach to understanding how tendons are constructed will inform future work to engineer improved tensile load-bearing tissues.

Concave-to-convex curve conversion of fiber cells correlates with Y-shaped suture formation at the poles of the rodent lens

The eye lens contains convexly curved fiber cells that align in concentric layers around the lens anterior-posterior pole axis. For lens fiber differentiation at the equator, cells elongate with their apical and basal tips migrating towards the anterior and posterior poles, respectively. At each pole, the fiber tips meet opposing tips of other fiber cells, to form a suture. Although umbilical or point sutures are observed in fish and birds, line, Y- or star-shaped sutures are detected in other vertebrate lenses. Sutures that do not converge at the point are thought to result from intricate movements of the fiber tips, rather than a straightforward migration along a meridional path. The triggers that give rise to these variations are currently not understood. Our findings revealed that in the mouse embryo, the early-stage lens contains only concave curved fibers, and later, a zone of concave-to-convex curve conversion develops. At this point, a nascent suture in a linear shape appears at the posterior pole and subsequently progresses into a V-shape. This V-shape appears to further develop into a Y-shape as a branch extends from the apex of the V-shape. In lens of zebrafish and Xenopus larvae that form point sutures, this curve-conversion zone is not observed. In lens of adult birds (e.g. zebra finch) that form a point suture, these too also lack a curve-conversion zone. In our previous studies, we demonstrated that murine lens fibers undergoing curve conversion extend membrane protrusions, or lamellipodia, at their basal membranes. In line with this, we did not observe protrusions at the basal tips of fibers in the non-mammalian lenses of zebrafish, Xenopus, and zebra finch in which curve conversion does not occur. We propose that the concave-to-convex conversion in rodent lenses introduces defined paths for fiber cell tips, leading to a more elaborate and complex suture formation, compared to the simple point suture of lower vertebrates.

Bionic structural design and model calculation for high toughness Si3N4f/BNi fibrous monolithic ceramic: A strategy from the fibrous cell and interface

Preeminent mechanical and dielectric performances are demonstrated by covalently bonded Si3N4 ceramics, yet their application range is still challenged because of limited toughness and sintering density. In this work, the Si3N4f/BNi fibrous monolithic ceramic with uniaxially arranged Si3N4 fibers cells separated by BN interfaces were successfully synthesized via hot-pressed sintering based on bionic structural design. The Si3N4f/BNi fibrous monolithic ceramics with bionic structure exhibited flexural strength, fracture toughness, and density of up to 501 MPa, 10.99 MPa m1/2, and 3.43 g/cm3, respectively. The results revealed that bionic structural design enables to significantly increase the toughness of the ceramic without sacrificing its strength. Furthermore, an original model for determining the flexural strength of fibrous monolithic ceramics was proposed, which can account for various shapes of fibrous cells. The findings of this research open up new prospects for achieving competitive mechanical performance of ceramics.

Membrane Interactions of GET1 and GET2 Facilitate Fiber Cell Initiation through the Guided Entry of the TA Protein Pathway in Cotton.

The guided entry of TA proteins (GET) pathway, which is responsible for the post-translational targeting and insertion of the tail-anchored (TA) protein into the endoplasmic reticulum (ER), plays an important role in physiological processes such as protein sorting, vesicle trafficking, cell apoptosis, and enzymatic reactions in which the GET1/2 complex is indispensable. However, a comprehensive study of the GET1 and GET2 genes and the GET pathway in cotton has not yet been carried out. Here, 12 GET1 and 21 GET2 genes were identified in nine representative plant species, and the phylogenetic relationships, gene structures, protein motifs, cis-regulatory elements (CREs), and temporal and spatial expression profiles were analyzed thoroughly. Our study indicated that GhGET1s and GhGET2s might be localized on ER membranes. According to expression profiling and CREs analysis, GhGET2-A02 was identified as a promising candidate for fiber cell development, interacting with two GhGET1s in the membrane, with a binding bias toward GhGET1-A06. Silencing of GhGET1-A06 or GhGET2-A02 reduced fiber initiation and elongation. In summary, our research provides important evidence for understanding the gene families and functions of GET1 and GET2 in cotton and provides clues for molecular breeding of high-quality cotton fiber varieties.

Canonical ligand-dependent and non-canonical ligand-independent EphA2 signaling in the eye lens of wild-type, knockout, and aging mice.

Disruption of Eph-ephrin bidirectional signaling leads to human congenital and age-related cataracts, but the mechanisms for these opacities in the eye lens remain unclear. Eph receptors bind to ephrin ligands on neighboring cells to induce canonical ligand-mediated signaling. The EphA2 receptor also signals non-canonically without ligand binding in cancerous cells, leading to epithelial-to-mesenchymal transition (EMT). We have previously shown that the receptor EphA2 and the ligand ephrin-A5 have diverse functions in maintaining lens transparency in mice. Loss of ephrin-A5 leads to anterior cataracts due to EMT. Surprisingly, both canonical and non-canonical EphA2 activation are present in normal wild-type lenses and in the ephrin-A5 knockout lenses. Canonical EphA2 signaling is localized exclusively to lens epithelial cells and does not change with age. Non-canonical EphA2 signaling is in both epithelial and fiber cells and increases significantly with age. We hypothesize that canonical ligand-dependent EphA2 signaling is required for the morphogenesis and organization of hexagonal equatorial epithelial cells while non-canonical ligand-independent EphA2 signaling is needed for complex membrane interdigitations that change during fiber cell differentiation and maturation. This is the first demonstration of non-canonical EphA2 activation in a non-cancerous tissue or cell and suggests a possible physiological function for ligand-independent EphA2 signaling.

Daily glycome and transcriptome profiling reveals polysaccharide structures and correlated glycosyltransferases critical for cotton fiber growth.

Cotton fiber is the most valuable naturally available material for the textile industry and the fiber length and strength are key determinants of its quality. Dynamic changes in the pectin, xyloglucan, xylan, and cellulose polysaccharide epitope content during fiber growth contribute to complex remodeling of fiber cell wall (CW) and quality. Detailed knowledge about polysaccharide compositional and structural alteration in the fiber during fiber elongation and strengthening is important to understand the molecular dynamics of fiber development and improve its quality. Here, large-scale glycome profiling coupled with fiber phenotype and transcriptome profiling was conducted on fiber collected daily covering the most critical window of fiber development. The profiling studies with high temporal resolution allowed us to identify specific polysaccharide epitopes associated with distinct fiber phenotypes that might contribute to fiber quality. This study revealed the critical role of highly branched RG-I pectin epitopes such as β-1,4-linked-galactans, β-1,6-linked-galactans, and arabinogalactans, in addition to earlier reported homogalacturonans and xyloglucans in the formation of cotton fiber middle lamella and contributing to fiber plasticity and elongation. We also propose the essential role of heteroxylans (Xyl-MeGlcA and Xyl-3Ar), as a guiding factor for secondary CW cellulose microfibril arrangement, thus contributing to fiber strength. Correlation analysis of profiles of polysaccharide epitopes from glycome data and expression profiles of glycosyltransferase-encoding genes from transcriptome data identified several key putative glycosyltransferases that are potentially involved in synthesizing the critical polysaccharide epitopes. The findings of this study provide a foundation to identify molecular factors that dictate important fiber traits.

Shc1 cooperates with Frs2 and Shp2 to recruit Grb2 in FGF-induced lens development.

Fibroblast growth factor (FGF) signaling elicits multiple downstream pathways, most notably the Ras/MAPK cascade facilitated by the adaptor protein Grb2. However, the mechanism by which Grb2 is recruited to the FGF signaling complex remains unresolved. Here we showed that genetic ablation of FGF signaling prevented lens induction by disrupting transcriptional regulation and actin cytoskeletal arrangements, which could be reproduced by deleting the juxtamembrane region of the FGF receptor and rescued by Kras activation. Conversely, mutations affecting the Frs2-binding site on the FGF receptor or the deletion of Frs2 and Shp2 primarily impact later stages of lens vesicle development involving lens fiber cell differentiation. Our study further revealed that the loss of Grb2 abolished MAPK signaling, resulting in a profound arrest of lens development. However, disrupting the Grb2 binding site on Shp2 or abrogating Shp2 phosphatase activity only modestly influenced FGF signaling, whereas mutating the presumed Shp2 dephosphorylation site on Grb2 did not impede MAPK signaling in lens development, indicating that Shp2 is only partially responsible for Grb2 recruitment. In contrast, we observed that FGF signaling is required for the phosphorylation of the Grb2-binding sites on Shc1 and the deletion of Shc1 exacerbates the lens vesicle defect caused by Frs2 and Shp2 deletion. These results reveal that Shc1 collaborates with Frs2 and Shp2 to target Grb2 in FGF signaling.

GhGRF4/GhARF2-GhGASA24 module regulates fiber cell wall thickness by modulating cellulose biosynthesis in upland cotton (Gossypium hirsutum).

Fiber elongation rate is an essential characteristic of cotton fiber in the textile industry, yet it has been largely overlooked in genetic studies. Gibberellins (GAs) and auxin (IAA) are recognized for their role in directing numerous developmental processes in plants by influencing cell differentiation and elongation. However, the degree to which GA-IAA interaction governs cellular elongation in cotton fiber cells remains to be fully understood. In this study, we identified a causal gene, Gibberellic Acid-Stimulated in Arabidopsis 24 (GhGASA24), that appears to be responsible for fiber elongation rate via regulating fiber cell wall thickness. Subsequent experiments revealed that GhGASA24 influences cell wall formation by promoting the expression of GhCesA8 and GhCesA10. Our findings suggest that Auxin Response Factor 2 (GhARF2) regulates fiber elongation rate by directly binding to the AuxRE elements in GhGASA24 promoter. In addition, we identified Growth Regulation Factor 4 (GhGRF4) as a transcription factor that interacts with GhARF2 to form a heterodimer complex, which also transcriptionally activates GhGASA24. Intriguingly, GhGRF4 regulates GhARF2 expression by directly binding to its promoter, thereby acting as a cascade regulator to enhance the transcriptional levels of GhGASA24. We propose that the GhGRF4/GhARF2-GhGASA24-GhCesAs module may contribute to fiber cell wall thickness by modulating cellulose biosynthesis, and provide a theoretical basis for improvement of fiber quality.

A New Pattern of Phytomelanin Deposition in Cypselae of Bahieae (Asteraceae, Heliantheae Alliance)

ABSTRACTPhytomelanin is the dark and highly inert substance found in the fruit walls of the Heliantheae alliance clade within Asteroideae (Asteraceae). Phytomelanin deposition pattern in the cypselar wall was studied in two taxa of the Bahieae—Bahia pedata A. Gray and Palafoxia arida B. L. Turner. The pericarp anatomy studies have revealed that the phytomelanin deposits in the schizogenous space between the hypodermis and fiber layer. Our findings reveal that phytomelanin deposition is associated with fiber and has also brought out a new pattern of phytomelanin deposition between inner (fibrous zone) and outer sclerified layers (hypodermal) in the schizogenous space. The study recorded a first‐time report of a second layer of phytomelanin outer to the sclerified hypodermal cells at maturity. Our observations point to phytomelanin precursors being synthesized as pliable fluid substances associated with fiber cells and solidifies later in the intercellular space. The study also recorded the first report for the occurrence of tracheoidal cells in the pericarp wall of the Bahieae within the Heliantheae alliance. Our study has provided insights to decipher evolutionary patterns and the significance of phytomelanin in the phytomelanic fruit clade (PFC), and it could help to resolve long‐standing debates on deposition patterns and the site of synthesis of phytomelanin.

Structural and anatomical analysis of rattan (Calamus balansaeanus Becc.)

Abstract Calamus balasaeanus Becc., a climbing palm species, has potential commercial value. Detailed anatomical and chemical analyses of rattan stems at different ages are crucial for their utilization in various industrial applications, particularly eco-friendly products. In this study, rattan stems of different ages were examined using light microscopy, transmission electron microscopy, and Fourier transform infrared spectroscopy. The results showed that the anatomical structure of rattan stems at different ages are very similar, with vascular bundle sizes ranging from 0.26 to 0.37 mm and a frequency of 8.4 to 13.7 bundles/mm2. The vessel lengths and diameters were 1.87–2.35 mm and 0.10–0.16 mm, respectively. Fiber lengths and diameters were 0.84–1.15 mm and 8.65–11.55 μm, respectively. As C. balasaeanus matured, the secondary wall of fiber cells thickened into the cell lumen, changing from 2 layers to 4 layers, the amount of crystallinity in cellulose increased while the mean microfibril angle decreased. The crystallinity of cellulose was higher in the periphery of the stem compared to the center. Unimodal imaging effectively visualized the distribution of various polysaccharides in C. balasaeanus tissues, with the highest concentrations of cellulose, hemicellulose, and lignin found in the base of the rattan stems.

Benzo[b]fluoranthene damages coronary artery and affects atherosclerosis markers in mice and umbilical vein endothelial cells

Polycyclic aromatic hydrocarbons (PAHs) exposure is associated with cardiovascular diseases. Toxic effects of PAHs are diverse, while cardiovascular consequences of benzo[b]fluoranthene (B[b]F) are unclear. Here, we reported the impacts of B[b]F on coronary artery and atherosclerosis markers both in mice and umbilical vein endothelial EAhy.926 cells. In mice, we found that B[b]F decreases heart-to-body weight ratio, affects aortic physiology, elevates serum low-density lipoprotein and total cholesterol, increases aortic levels of collagen fiber and atherosclerotic marker vascular cell adhesion molecule-1 (VCAM-1), and downregulates oxidative stress related nuclear factor erythroid 2-related factor 2 (Nrf2). In EAhy.926 cells, we showed that B[b]F inhibits cell proliferation and migration in a dose-dependent manner, induces cell cycle arrest and apoptosis, increases reactive oxygen species, upregulates VCAM-1 level, and suppresses expression of Nrf2. Taken together, our findings reveal that B[b]F exposure may contribute to coronary artery damage and potentially induce atherosclerosis, possibly via the Nrf2-related signaling pathways.

Mechanism of low-intensity pulse ultrasound combined with Rhodiola to promote bone formation in spinal fusion.

This study aimed to explore the influence and mechanism of low-intensity pulsed ultrasound (LIPUS) combined with Rhodiola bone penetration on the formation of spinal fusion bone. Sixty clean-grade New Zealand white rabbits were selected for randomization and divided into combined group and Rhodiola group, with 30 rabbits in each group to construct a rabbit lumbar intervertebral fusion model, using Rhodiola intervention and Rhodiola combined with LIPUS intervention protocol, respectively. The axial strength, axial stiffness, maximum compressive load, vascular endothelial growth factor (VEGF), cycloxygenase-2 (COX-2), prostaglandin E2 (PGE2) and transforming growth factor-β (TGF-β) were compared after HE staining, immunohistochemistry and biomechanical detection. Spine fusion rate was 100.00%; the combined bone graft tissue had implanted bone cell degeneration, cell necrosis and cell hyperplasia, chondrocytes differentiated into trabecular bone and some hematopoietic cells, severe cell necrosis and fiber cell proliferation and late bone formation in the Rhodiola group, VEGF, COX-2, PGE2, TGF-β, axial strength, axial stiffness, and maximum compression load in the combined group significantly increased (P<0.05). Spinal fusion using LIPUS combined with Rhodiola can enhance biomechanical properties and promote the role of PGE2, COX-2, VEGF, TGF-β expression and bone formation, and this protocol is worthy of clinical application.

Characterization and property prediction of fibre structures within discontinuous-fibre reinforced polymer matrix composites using 3D fibre cells assisted by contrastive learning

Fibre-cell-based fibre structure characterization approach was proposed recently to characterize the fibre distribution within discontinuous-fibre reinforced polymer matrix composites (DFR PMCs) over a 2D domain. This approach determines the distribution state of each fibre based on the relative size and topological features of its fibre cell. In this study, the fibre-cell-based approach is extended for 3D fibre domains. A convolutional neural network (CNN) encoder is trained through contrastive learning to quantitatively represent topological features of 3D fibre cells. Subsequently, the feature–property correlations are established using an artificial neural network (ANN). For practical application, the ANN is integrated with an image analysis software to provide in situ predictions of local elastic modulus of a DFR PMC based on its fibre structures observed from micro-CT images. The predictions are also compared with the experimental measurements acquired through microindentation testing, and it shows a good agreement.

PagNAC2a promotes phloem fiber development by regulating PagATL2 in poplar

Phloem fiber is a key component of phloem tissue and is involved in supporting its structural integrity. NAC domain transcription factors are master switches that regulate secondary cell wall (SCW) biosynthesis in xylem fibers, but the mechanism by which NACs regulate phloem fiber development remains unexplored. Here, a NAC2-like gene in poplar, PagNAC2a, was shown to be involved in phloem fiber differentiation. qRT-PCR and GUS staining revealed that PagNAC2a was specifically expressed in the phloem zone of poplar stems. The overexpression of PagNAC2a in poplar increased plant biomass by increasing plant height, stem diameter, and leaf area. Stem anatomy analysis revealed that overexpression of PagNAC2a resulted in enhanced phloem fiber differentiation and cell wall deposition. In addition, PagNAC2a directly upregulated the expression of PagATL2, a gene involved in phloem development, as revealed by yeast one hybrid (Y1H) and electrophoretic mobility shift assay (EMSA) assays. Overall, we proposed that the PagNAC2a was a positive regulator of phloem fiber development in poplar, and these results provided insights into the molecular mechanisms involved in the differentiation of phloem fibers.

Mode II fracture properties of parallel neosinocalamus affinis bamboo strand lumber

This study investigates the Mode II fracture properties in the RL and TL planes of parallel neosinocalamus affinis bamboo strand lumber. Eighty End-Notched Flexure (ENF) specimens were designed, varying pre-crack lengths and specimen widths. Digital Image Correlation (DIC) was employed to analyze strain variations during the development of the fracture process zone (FPZ) and crack propagation stages. The Compliance Based Beam Method (CBBM) and equivalent crack length calculations were used to determine the strain energy release rate (GIIC) of PNABSL. R curves and crack growth rate curves revealed that crack propagation in the RL and TL planes. The results indicated that Mode II fracture propagation in both the RL and TL planes of PNABSL predominantly exhibited self-similar cracking, though the fracture surfaces were rough and uneven, with noticeable fiber bridging. The study found that Mode II fracture propagation was less stable in the RL plane compared to the TL plane, which is attributed to the easier fracturing of thick-walled fiber cells in the RL plane. The critical strain energy release rate for PNABSL was determined, offering valuable insights into its fracture behavior.

Effects of Supplementation with Essential Fatty Acids and Conjugated Linoleic Acids on Muscle Structure and Fat Deposition in Lactating Holstein Cows

High-yielding dairy cows need diets that meet their energy demand and contain sufficient essential nutrients such as n-3 fatty acids (FAs). Conjugated linoleic acid (CLA) is able to relieve the energy metabolism, but common corn silage and concentrate-based diets contain insufficient amounts of essential fatty acids (EFA). Abomasal infusion was used in the current study to supplement cows from 9 weeks antepartum to 9 weeks postpartum with either coconut oil (CTRL, n = 8), EFA (n = 9), or conjugated linoleic acid (CLA, n = 9), or a combination of both (EFA + CLA, n = 10). The study focused on the effects of FAs on peripheral tissues, such as longissimus muscle (MLD) and adipose tissues, which were harvested after slaughter. Fatty acid composition, muscle fiber and fat cell morphology, muscle fiber type transition, and gene expression were analyzed. Supplemented FAs and their metabolites were increased (p &lt; 0.05) in MLD and intermuscular fat (INTF) but not in subcutaneous fat (SCF). The intramuscular fat content and gene expression of ACACA and FASN were increased in CLA-supplemented cows (p &lt; 0.05). Supplementation did not affect the muscle fiber size and fiber type composition. Supplemented CLA had more effects than EFA, improving the energy balance of cows accompanied with increased triglyceride formation and storage.

Transient Notch Activation Converts Pluripotent Stem Cell-Derived Cardiomyocytes Towards a Purkinje Fiber Fate.

Cardiac Purkinje fibers form the most distal part of the ventricular conduction system. They coordinate contraction and play a key role in ventricular arrhythmias. While many cardiac cell types can be generated from human pluripotent stem cells, methods to generate Purkinje fiber cells remain limited, hampering our understanding of Purkinje fiber biology and conduction system defects. To identify signaling pathways involved in Purkinje fiber formation, we analyzed single cell data from murine embryonic hearts and compared Purkinje fiber cells to trabecular cardiomyocytes. This identified several genes, processes, and signaling pathways putatively involved in cardiac conduction, including Notch signaling. We next tested whether Notch activation could convert human pluripotent stem cell-derived cardiomyocytes to Purkinje fiber cells. Following Notch activation, cardiomyocytes adopted an elongated morphology and displayed altered electrophysiological properties including increases in conduction velocity, spike slope, and action potential duration, all characteristic features of Purkinje fiber cells. RNA-sequencing demonstrated that Notch-activated cardiomyocytes undergo a sequential transcriptome shift, which included upregulation of key Purkinje fiber marker genes involved in fast conduction such as SCN5A, HCN4 and ID2, and downregulation of genes involved in contractile maturation. Correspondingly, we demonstrate that Notch-induced cardiomyocytes have decreased contractile force in bioengineered tissues compared to control cardiomyocytes. We next modified existing in silico models of human pluripotent stem cell-derived cardiomyocytes using our transcriptomic data and modeled the effect of several anti-arrhythmogenic drugs on action potential and calcium transient waveforms. Our models predicted that Purkinje fiber cells respond more strongly to dofetilide and amiodarone, while cardiomyocytes are more sensitive to treatment with nifedipine. We validated these findings in vitro , demonstrating that our new cell-specific in vitro model can be utilized to better understand human Purkinje fiber physiology and its relevance to disease.

A process-based model of climate-driven xylogenesis and tree-ring formation in broad-leaved trees (BTR).

The process-based xylem formation model is an important tool for understanding the radial growth process of trees and its influencing factors. While numerous xylogenesis models for conifers have been developed, there is a lack of models available for non-coniferous trees. In this study, we present a process-based model designed for xylem formation and ring growth in broad-leaved trees, which we call the Broad-leaved Tree-Ring (BTR) model. Climate factors, including day length, air temperature, soil moisture, and vapor pressure deficit, drive daily xylem cell production (fibers and vessels) and growth (enlargement, wall deposition). The model calculates the total cell area in the simulated zone to determine the annual ring width. The results demonstrate that the BTR model can basically simulate inter-annual variation in ring width and intra-annual changes in vessel and fiber cell formation in Fraxinus mandshurica (ring-porous) and Betula platyphylla (diffuse-porous). The BTR model is a potential tool for understanding how different trees form wood and how climate change influences this process.

GhCASPL1 regulates secondary cell wall thickening in cotton fibers by stabilizing the cellulose synthase complex on the plasma membrane.

Cotton (Gossypium hirsutum) fibers are elongated single cells that rapidly accumulate cellulose during secondary cell wall (SCW) thickening, which requires cellulose synthase complex (CSC) activity. Here, we describe the CSC-interacting factor CASPARIAN STRIP MEMBRANE DOMAIN-LIKE1 (GhCASPL1), which contributes to SCW thickening by influencing CSC stability on the plasma membrane. GhCASPL1 is preferentially expressed in fiber cells during SCW biosynthesis and encodes a MARVEL domain protein. The ghcaspl1 ghcaspl2 mutant exhibited reduced plant height and produced mature fibers with fewer natural twists, lower tensile strength, and a thinner SCW compared to the wild type. Similarly, the Arabidopsis (Arabidopsis thaliana) caspl1 caspl2 double mutant showed a lower cellulose content and thinner cell walls in the stem vasculature than the wild type but normal plant morphology. Introducing the cotton gene GhCASPL1 successfully restored the reduced cellulose content of the Arabidopsis caspl1 caspl2 mutant. Detergent treatments, ultracentrifugation assays, and enzymatic assays showed that the CSC in the ghcaspl1 ghcaspl2 double mutant showed reduced membrane binding and decreased enzyme activity compared to the wild type. GhCASPL1 binds strongly to phosphatidic acid (PA), which is present in much higher amounts in thickening fiber cells compared to ovules and leaves. Mutating the PA-binding site in GhCASPL1 resulted in the loss of its colocalization with GhCesA8, and it failed to localize to the plasma membrane. PA may alter membrane structure to facilitate protein-protein interactions, suggesting that GhCASPL1 and PA collaboratively stabilize the CSC. Our findings shed light on CASPL functions and the molecular machinery behind SCW biosynthesis in cotton fibers.