RESUMEN
The stereotypical tip growth of filamentous fungi supports their lifestyles and functions. It relies on the polarized remodeling and expansion of a protective elastic cell wall (CW) driven by large cytoplasmic turgor pressure. Remarkably, hyphal filament diameters and cell elongation rates can vary extensively among different fungi. To date, however, how fungal cell mechanics may be adapted to support these morphological diversities while ensuring surface integrity remains unknown. Here, we combined super-resolution imaging and deflation assays to measure local CW thickness, elasticity and turgor in a set of fungal species spread on the evolutionary tree that spans a large range in cell size and growth speeds. While CW elasticity exhibited dispersed values, presumably reflecting differences in CW composition, both thickness and turgor scaled in dose-dependence with cell diameter and growth speeds. Notably, larger cells exhibited thinner lateral CWs, and faster cells thinner apical CWs. Counterintuitively, turgor pressure was also inversely scaled with cell diameter and tip growth speed, challenging the idea that turgor is the primary factor dictating tip elongation rates. We propose that fast-growing cells with rapid CW turnover have evolved strategies based on a less turgid cytoplasm and thin walls to safeguard surface integrity and survival.
Asunto(s)
Pared Celular , Hongos , Hifa , Pared Celular/metabolismo , Pared Celular/fisiología , Hifa/crecimiento & desarrollo , Hongos/fisiología , Elasticidad , Citoplasma/metabolismo , Fenómenos BiomecánicosRESUMEN
BACKGROUND: Like all plant cells, the guard cells of stomatal complexes are encased in cell walls that are composed of diverse, interacting networks of polysaccharide polymers. The properties of these cell walls underpin the dynamic deformations that occur in guard cells as they expand and contract to drive the opening and closing of the stomatal pore, the regulation of which is crucial for photosynthesis and water transport in plants. SCOPE: Our understanding of how cell wall mechanics are influenced by the nanoscale assembly of cell wall polymers in guard cell walls, how this architecture changes over stomatal development, maturation and ageing and how the cell walls of stomatal guard cells might be tuned to optimize stomatal responses to dynamic environmental stimuli is still in its infancy. CONCLUSION: In this review, we discuss advances in our ability to probe experimentally and to model the structure and dynamics of guard cell walls quantitatively across a range of plant species, highlighting new ideas and exciting opportunities for further research into these actively moving plant cells.
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Pared Celular , Estomas de Plantas , Poaceae , Pared Celular/metabolismo , Pared Celular/fisiología , Estomas de Plantas/fisiología , Poaceae/fisiología , Poaceae/crecimiento & desarrolloRESUMEN
Multicellular organisms grow and acquire their shapes through the differential expansion and deformation of their cells. Recent research has addressed the role of cell and tissue mechanical properties in these processes. In plants, it is believed that growth rate is a function of the mechanical stress exerted on the cell wall, the thin polymeric layer surrounding cells, involving an effective viscosity. Nevertheless, recent studies have questioned this view, suggesting that cell wall elasticity sets the growth rate or that uptake of water is limiting for plant growth. To assess these issues, we developed a microfluidic device to quantify the growth rates, elastic properties and hydraulic conductivity of individual Marchantia polymorpha plants in a controlled environment with a high throughput. We characterized the effect of osmotic treatment and abscisic acid on growth and hydromechanical properties. Overall, the instantaneous growth rate of individuals is correlated with both bulk elastic modulus and hydraulic conductivity. Our results are consistent with a framework in which the growth rate is determined primarily by the elasticity of the wall and its remodelling, and secondarily by hydraulic conductivity. Accordingly, the coupling between the chemistry of the cell wall and the hydromechanics of the cell appears as key to set growth patterns during morphogenesis.
Asunto(s)
Pared Celular , Pared Celular/fisiología , Marchantia/crecimiento & desarrollo , Marchantia/fisiología , Ácido Abscísico/metabolismo , Modelos Biológicos , Fenómenos Biomecánicos , Desarrollo de la Planta/fisiologíaRESUMEN
Stomata are pores at the leaf surface that enable gas exchange and transpiration. The signaling pathways that regulate the differentiation of stomatal guard cells and the mechanisms of stomatal pore formation have been characterized in Arabidopsis thaliana. However, the process by which stomatal complexes develop after pore formation into fully mature complexes is poorly understood. We tracked the morphogenesis of young stomatal complexes over time to establish characteristic geometric milestones along the path of stomatal maturation. Using 3D-nanoindentation coupled with finite element modeling of young and mature stomata, we found that despite having thicker cell walls than young guard cells, mature guard cells are more energy efficient with respect to stomatal opening, potentially attributable to the increased mechanical anisotropy of their cell walls and smaller changes in turgor pressure between the closed and open states. Comparing geometric changes in young and mature guard cells of wild-type and cellulose-deficient plants revealed that although cellulose is required for normal stomatal maturation, mechanical anisotropy appears to be achieved by the collective influence of cellulose and additional wall components. Together, these data elucidate the dynamic geometric and biomechanical mechanisms underlying the development process of stomatal maturation.
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Arabidopsis , Pared Celular , Estomas de Plantas , Arabidopsis/fisiología , Arabidopsis/crecimiento & desarrollo , Arabidopsis/genética , Estomas de Plantas/fisiología , Estomas de Plantas/crecimiento & desarrollo , Estomas de Plantas/citología , Anisotropía , Pared Celular/metabolismo , Pared Celular/fisiología , Celulosa/metabolismo , Análisis de Elementos Finitos , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genéticaRESUMEN
Sorghum stems comprise different tissue components, i.e., rind, pith, and vascular bundles in the rind and pith regions, of different cell morphologies and cell wall characteristics. The overall responses of stems to mechanical loadings depend on the responses of these tissues themselves. Investigating how each tissue deforms to various loading conditions will inform us of the failure mechanisms in sorghum stems when exposed to wind loadings, which can guide the development of lodging-resistant variants. To this end, numerical analyses were implemented to investigate the effects of cell morphologies and cell wall properties on the overall mechanical responses of the above four tissues under tension and compression. Microstructures of different tissues were constructed from microscopic images of the tissues using computer-aided design (CAD), which were then used for finite element (FE) analyses. Shell finite elements were used to model the cell walls, and the classical lamination model was used to determine the overall mechanical responses of cell walls having different fiber composite arrangements. The results from the numerical analyses helped explain how the loading (boundary) conditions, the cell microstructures, the mechanical properties of cell walls of different tissues, the cell wall thickness, the microfibril angle (MFA) of fiber composites of the cell walls, and the turgor pressure affected the overall mechanical responses of the tissues. Tissue stiffening or softening behaviors were attributed to different microstructural deformations, i.e., local or global buckling of cell walls, cell collapse, densifications of cells, or reorientation and rearrangement of cells. The mechanical properties and thickness of cell walls only affected the stiffness and load-bearing ability of the tissues. The turgor pressure affected the compressive responses but its effect on tensile responses was negligible. The MFA had a significant influence on the stiffness and load-bearing ability when the tissues were loaded along their longitudinal axis, but it had an insignificant effect on loading in the transverse direction. Tissues with smaller cell sizes and denser cells were stronger and stiffer than those with larger cell sizes. The numerical simulations also revealed that rind and rind vascular bundles were stiffer and had higher load-bearing ability than pith and pith vascular bundles.
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Sorghum , Estrés Mecánico , Presión , Pared Celular/fisiología , Análisis de Elementos FinitosRESUMEN
The mechanical properties of guard cells have major effects on stomatal functioning. Reinforced stiffness in the stomatal polar regions was recently proposed to play an important role in stomatal function, but the underlying molecular mechanisms remain elusive. Here, we used genetic and biochemical approaches in poplar (Populus spp.) to show that the transcription factor MYB156 controls pectic homogalacturonan-based polar stiffening through the downregulation of the gene encoding pectin methylesterase 6 (PME6). Loss of MYB156 increased the polar stiffness of stomata, thereby enhancing stomatal dynamics and response speed to various stimuli. In contrast, overexpression of MYB156 resulted in decreased polar stiffness and impaired stomatal dynamics, accompanied by smaller leaves. Polar stiffening functions in guard cell dynamics in response to changing environmental conditions by maintaining normal stomatal morphology during stomatal movement. Our study revealed the structure-function relationship of the cell wall of guard cells in stomatal dynamics, providing an important means for improving the stomatal performance and drought tolerance of plants.
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Estomas de Plantas , Populus , Estomas de Plantas/fisiología , Factores de Transcripción/genética , Populus/genética , Regulación de la Expresión Génica de las Plantas/genética , Pared Celular/fisiologíaRESUMEN
The fungal cell is surrounded by a thick cell wall which obviously play an essential role in the protection of the fungus against external aggressive environments. In spite of 50 years of studies, the cell wall remains poorly known and especially its constant modifications during growth as well as environmental changes is not well appreciated. This review focus on the cell wall changes seen between different fungal stages and cell populations with a specific view to explain the resistance to stresses.
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Aspergillus fumigatus , Proteínas Fúngicas , Virulencia , Pared Celular/fisiologíaRESUMEN
Most bacteria are surrounded by a cell wall composed of peptidoglycan (PG) that specifies shape and protects the cell from osmotic rupture. Growth, division, and morphogenesis are intimately linked to the synthesis of this exoskeleton but also its hydrolysis. The enzymes that cleave the PG meshwork require careful control to prevent aberrant hydrolysis and loss of envelope integrity. Bacteria employ diverse mechanisms to control the activity, localization, and abundance of these potentially autolytic enzymes. Here, we discuss four examples of how cells integrate these control mechanisms to finely tune cell wall hydrolysis. We highlight recent advances and exciting avenues for future investigation.
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N-Acetil Muramoil-L-Alanina Amidasa , Peptidoglicano , Bacterias , Pared Celular/fisiología , Proteínas BacterianasRESUMEN
The cell wall (CW) is a thin and rigid layer encasing the membrane of all plant and fungal cells. It ensures mechanical integrity by bearing mechanical stresses derived from large cytoplasmic turgor pressure, contacts with growing neighbors or growth within restricted spaces. The CW is made of polysaccharides and proteins, but is dynamic in nature, changing composition and geometry during growth, reproduction or infection. Such continuous and often rapid remodeling entails risks of enhanced stress and consequent damages or fractures, raising the question of how the CW detects and measures surface mechanical stress and how it strengthens to ensure surface integrity? Although early studies in model fungal and plant cells have identified homeostatic pathways required for CW integrity, recent methodologies are now allowing the measurement of pressure and local mechanical properties of CWs in live cells, as well as addressing how forces and stresses can be detected at the CW surface, fostering the emergence of the field of CW mechanobiology. Here, using tip-growing cells of plants and fungi as case study models, we review recent progress on CW mechanosensation and mechanical regulation, and their implications for the control of cell growth, morphogenesis and survival.
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Pared Celular , Células Vegetales , Pared Celular/fisiología , Morfogénesis , Estrés Mecánico , BiofisicaRESUMEN
The stiffness of a plant cell in response to an applied force is determined not only by the elasticity of the cell wall but also by turgor pressure and cell geometry, which affect the tension of the cell wall. Although stiffness has been investigated using atomic force microscopy (AFM) and Young's modulus of the cell wall has occasionally been estimated using the contact-stress theory (Hertz theory), the existence of tension has made the study of stiffness more complex. Elastic shell theory has been proposed as an alternative method; however, the estimation of elasticity remains ambiguous. Here, we used finite element method simulations to verify the formula of the elastic shell theory for onion (Allium cepa) cells. We applied the formula and simulations to successfully quantify the turgor pressure and elasticity of a cell in the plane direction using the cell curvature and apparent stiffness measured by AFM. We conclude that tension resulting from turgor pressure regulates cell stiffness, which can be modified by a slight adjustment of turgor pressure in the order of 0.1 MPa. This theoretical analysis reveals a path for understanding forces inherent in plant cells.
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Pared Celular , Células Vegetales , Pared Celular/fisiología , Módulo de Elasticidad , Elasticidad , Microscopía de Fuerza Atómica/métodos , Cebollas , Células Vegetales/fisiologíaRESUMEN
GTP is an important signaling molecule involved in the growth, development, and stress adaptability of plants. The functions are mediated via binding to GTPases which are in turn regulated by GTPase-activating proteins (GAPs). Satellite reports have suggested the positive roles of GAPs in regulating ABA signaling and pathogen resistance in plants. However, the molecular mechanisms that bring forth the pathogen resistance have remained unclear. In this study, we demonstrated that the expression of AtGAP1 was inducible by Pseudomonas syringae pv. tomato DC3000 (Pst DC3000). The overexpression of AtGAP1 in Arabidopsis promoted the expression of PR1 and the resistance to Pst DC3000. Proteomic analyses revealed the enhanced accumulation of cell-wall-modifying proteins as a result of AtGAP1 overexpression. By microscopic analyses, we showed that the overexpression of AtGAP1 resulted in increased thickness of the mesophyll cell wall and reduced stomatal aperture, which are effective strategies for restricting the entry of foliar pathogens. Altogether, we demonstrated that AtGAP1 increases the resistance to Pst DC3000 in Arabidopsis by promoting cellular strategies that restrict the entry of pathogens into the cells. These results point to a future direction for studying the modes of action of GAPs in regulating plant cell structures and disease resistance.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Proteínas Activadoras de GTPasa , Pseudomonas syringae , Solanum lycopersicum , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Pared Celular/fisiología , Resistencia a la Enfermedad/genética , Proteínas Activadoras de GTPasa/genética , Proteínas Activadoras de GTPasa/metabolismo , Regulación de la Expresión Génica de las Plantas , Solanum lycopersicum/metabolismo , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Estomas de Plantas/fisiología , Proteómica , Pseudomonas syringae/fisiologíaRESUMEN
Plant cell walls are highly dynamic and chemically complex structures surrounding all plant cells. They provide structural support, protection from both abiotic and biotic stress as well as ensure containment of turgor. Recently evidence has accumulated that a dedicated mechanism exists in plants, which is monitoring the functional integrity of cell walls and initiates adaptive responses to maintain integrity in case it is impaired during growth, development or exposure to biotic and abiotic stress. The available evidence indicates that detection of impairment involves mechano-perception, while reactive oxygen species and phytohormone-based signaling processes play key roles in translating signals generated and regulating adaptive responses. More recently it has also become obvious that the mechanisms mediating cell wall integrity maintenance and pattern triggered immunity are interacting with each other to modulate the adaptive responses to biotic stress and cell wall integrity impairment. Here we will review initially our current knowledge regarding the mode of action of the maintenance mechanism, discuss mechanisms mediating responses to biotic stresses and highlight how both mechanisms may modulate adaptive responses. This first part will be focused on Arabidopsis thaliana since most of the relevant knowledge derives from this model organism. We will then proceed to provide perspective to what extent the relevant molecular mechanisms are conserved in other plant species and close by discussing current knowledge of the transcriptional machinery responsible for controlling the adaptive responses using selected examples.
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Arabidopsis , Transducción de Señal , Arabidopsis/genética , Pared Celular/fisiología , Regulación de la Expresión Génica de las Plantas , Células Vegetales/fisiología , Plantas , Estrés FisiológicoRESUMEN
Cell growth in plants occurs due to relaxation of the cell wall in response to mechanical forces generated by turgor pressure. Growth can be anisotropic, with the principal direction of growth often correlating with the direction of lower stiffness of the cell wall. However, extensometer experiments on onion epidermal peels have shown that the tissue is stiffer in the principal direction of growth. Here, we used a combination of microextensometer experiments on epidermal onion peels and finite element method (FEM) modeling to investigate how cell geometry and cellular patterning affects mechanical measurements made at the tissue level. Simulations with isotropic cell-wall material parameters showed that the orientation of elongated cells influences tissue apparent stiffness, with the tissue appearing much softer in the transverse versus the longitudinal directions. Our simulations suggest that although extensometer experiments show that the onion tissue is stiffer when stretched in the longitudinal direction, the effect of cellular geometry means that the wall is in fact softer in this direction, matching the primary growth direction of the cells.
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Pared Celular , Fenómenos Mecánicos , Anisotropía , Pared Celular/fisiologíaRESUMEN
The instructive role of mechanical cues during morphogenesis is increasingly being recognized in all kingdoms. Patterns of mechanical stress depend on shape, growth and external factors. In plants, the cell wall integrates these three parameters to function as a hub for mechanical feedback. Plant cells are interconnected by cell walls that provide structural integrity and yet are flexible enough to act as both targets and transducers of mechanical cues. Such cues may act locally at the subcellular level or across entire tissues, requiring tight control of both cell-wall composition and cell-cell adhesion. Here we focus on how changes in cell-wall chemistry and mechanics act in communicating diverse cues to direct growth asymmetries required for plant morphogenesis. We explore the role of cellulose microfibrils, microtubule arrays and pectin methylesterification in the transduction of mechanical cues during morphogenesis. Plant hormones can affect the mechanochemical composition of the cell wall and, in turn, the cell wall can modulate hormone signaling pathways, as well as the tissue-level distribution of these hormones. This also leads us to revisit the position of biochemical growth factors, such as plant hormones, acting both upstream and downstream of mechanical signaling. Finally, while the structure of the cell wall is being elucidated with increasing precision, existing data clearly show that the integration of genetic, biochemical and theoretical studies will be essential for a better understanding of the role of the cell wall as a hub for the mechanical control of plant morphogenesis.
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Células Vegetales , Reguladores del Crecimiento de las Plantas , Pared Celular/fisiología , Morfogénesis , Células Vegetales/fisiología , Desarrollo de la Planta , Estrés MecánicoRESUMEN
Septation in filamentous fungi is a normal part of development, which involves the formation of cross-hyphal bulkheads, typically containing pores, allowing cytoplasmic streaming between compartments. Based on previous findings regarding septa and cell wall stress, we hypothesized that septa are critical for survival during cell wall stress. To test this hypothesis, we used known Aspergillus nidulans septation-deficient mutants (ΔsepH, Δbud3, Δbud4, and Δrho4) and six antifungal compounds. Three of these compounds (micafungin, Congo red, and calcofluor white) are known cell wall stressors which activate the cell wall integrity signaling pathway (CWIS), while the three others (cycloheximide, miconazole, and 2,3-butanedione monoxime) perturb specific cellular processes not explicitly related to the cell wall. Our results show that deficiencies in septation lead to fungi which are more susceptible to cell wall-perturbing compounds but are no more susceptible to other antifungal compounds than a control. This implies that septa play a critical role in surviving cell wall stress. IMPORTANCE The ability to compartmentalize potentially lethal damage via septation appears to provide filamentous fungi with a facile means to tolerate diverse forms of stress. However, it remains unknown whether this mechanism is deployed in response to all forms of stress or is limited to specific perturbations. Our results support the latter possibility by showing that presence of septa promotes survival in response to cell wall damage but plays no apparent role in coping with other unrelated forms of stress. Given that cell wall damage is a primary effect caused by exposure to the echinocandin class of antifungal agents, our results emphasize the important role that septa might play in enabling resistance to these drugs. Accordingly, the inhibition of septum formation could conceivably represent an attractive approach to potentiating the effects of echinocandins and mitigating resistance in human fungal pathogens.
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Aspergillus nidulans/crecimiento & desarrollo , Aspergillus nidulans/fisiología , Pared Celular/fisiología , Antifúngicos/farmacología , Aspergillus nidulans/efectos de los fármacos , Aspergillus nidulans/genética , Pared Celular/efectos de los fármacos , Pared Celular/genética , Rojo Congo/farmacología , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Hifa/efectos de los fármacos , Hifa/genética , Hifa/crecimiento & desarrollo , Hifa/metabolismo , Micafungina/farmacocinética , Viabilidad Microbiana/efectos de los fármacos , Estrés FisiológicoRESUMEN
Efficient uptake of nutrients in both animal and plant cells requires tissue-spanning diffusion barriers separating inner tissues from the outer lumen/soil. However, we poorly understand how such contiguous three-dimensional superstructures are formed in plants. Here, we show that correct establishment of the plant Casparian Strip (CS) network relies on local neighbor communication. We show that positioning of Casparian Strip membrane domains (CSDs) is tightly coordinated between neighbors in wild-type and that restriction of domain formation involves the putative extracellular protease LOTR1. Impaired domain restriction in lotr1 leads to fully functional CSDs at ectopic positions, forming 'half strips'. LOTR1 action in the endodermis requires its expression in the stele. LOTR1 endodermal expression cannot complement, while cortex expression causes a dominant-negative phenotype. Our findings establish LOTR1 as a crucial player in CSD positioning acting in a directional, non-cell-autonomous manner to restrict and coordinate CS positioning.
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Proteínas de Arabidopsis , Pared Celular , Lignina , Arabidopsis/citología , Arabidopsis/genética , Arabidopsis/fisiología , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Pared Celular/química , Pared Celular/metabolismo , Pared Celular/fisiología , Lignina/química , Lignina/genética , Lignina/fisiología , Regiones Promotoras Genéticas/genéticaRESUMEN
With their ability of CO2 fixation using sunlight as an energy source, algae and especially microalgae are moving into the focus for the production of proteins and other valuable compounds. However, the valorization of algal biomass depends on the effective disruption of the recalcitrant microalgal cell wall. Especially cell walls of Chlorella species proved to be very robust. The wall structures that are responsible for this robustness have been studied less so far. Here, we evaluate different common methods to break up the algal cell wall effectively and measure the success by protein and carbohydrate release. Subsequently, we investigate algal cell wall features playing a role in the wall's recalcitrance towards disruption. Using different mechanical and chemical technologies, alkali catalyzed hydrolysis of the Chlorella vulgaris cells proved to be especially effective in solubilizing up to 56 wt% protein and 14 wt% carbohydrates of the total biomass. The stepwise degradation of C. vulgaris cell walls using a series of chemicals with increasingly strong conditions revealed that each fraction released different ratios of proteins and carbohydrates. A detailed analysis of the monosaccharide composition of the cell wall extracted in each step identified possible factors for the robustness of the cell wall. In particular, the presence of chitin or chitin-like polymers was indicated by glucosamine found in strong alkali extracts. The presence of highly ordered starch or cellulose was indicated by glucose detected in strong acidic extracts. Our results might help to tailor more specific efforts to disrupt Chlorella cell walls and help to valorize microalgae biomass.
Asunto(s)
Pared Celular/química , Pared Celular/fisiología , Chlorella vulgaris/metabolismo , Biomasa , Carbohidratos/análisis , Pared Celular/patología , Celulosa/análisis , Chlorella vulgaris/patogenicidad , Microalgas/metabolismo , Almidón/análisisRESUMEN
BACKGROUND: Despite the frequent use of protoplast-to-plant system in in vitro cultures of plants, the molecular mechanisms regulating the first and most limiting stages of this process, i.e., protoplast dedifferentiation and the first divisions leading to the formation of a microcallus, have not been elucidated. RESULTS: In this study, we investigated the function of miRNAs in the dedifferentiation of A. thaliana mesophyll cells in a process stimulated by the enzymatic removal of the cell wall. Leaf cells, protoplasts and CDPs (cells derived from protoplasts) cultured for 24, 72 and 120 h (first cell division). In protoplasts, a strong decrease in the amount of AGO1 in both the nucleus and the cytoplasm, as well as dicing bodies (DBs), which are considered to be sites of miRNA biogenesis, was shown. However during CDPs division, the amounts of AGO1 and DBs strongly increased. MicroRNA transcriptome studies demonstrated that lower amount of differentially expressed miRNAs are present in protoplasts than in CDPs cultured for 120 h. Then analysis of differentially expressed miRNAs, selected pri-miRNA and mRNA targets were performed. CONCLUSION: This result indicates that miRNA function is not a major regulation of gene expression in the initial but in later steps of dedifferentiation during CDPs divisions. miRNAs participate in organogenesis, oxidative stress, nutrient deficiencies and cell cycle regulation in protoplasts and CDPs. The important role played by miRNAs in the process of dedifferentiation of mesophyll cells was confirmed by the increased mortality and reduced cell division of CDPs derived from mutants with defective miRNA biogenesis and miR319b expression.
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Arabidopsis/fisiología , Desdiferenciación Celular/genética , Pared Celular/fisiología , MicroARNs/genética , Células Vegetales/fisiología , ARN de Planta/genética , Arabidopsis/genética , MicroARNs/metabolismo , ARN de Planta/metabolismoRESUMEN
Cytokinesis is the final step of the cell-division cycle. In fungi, it relies on the coordination of constriction of an actomyosin contractile ring and construction of the septum at the division site. Glucan synthases synthesize glucans, which are the major components in fungal cell walls and division septa. It is known that Rho1 and Rho2 GTPases regulate glucan synthases Bgs1, Bgs4, and Ags1, and that Sbg1 and the F-BAR protein Cdc15 play roles in Bgs1 stability and delivery to the plasma membrane. Here we characterize Smi1, an intrinsically disordered protein that interacts with Bgs4 and regulates its trafficking and localization in fission yeast. Smi1 is important for septum integrity, and its absence causes severe lysis during cytokinesis. Smi1 localizes to secretory vesicles and moves together with Bgs4 toward the division site. The concentrations of the glucan synthases Bgs1 and Bgs4 and the glucanases Agn1 and Bgl2 decrease at the division site in the smi1 mutant, but Smi1 seems to be more specific to Bgs4. Mistargeting of Smi1 to mitochondria mislocalizes Bgs4 but not Bgs1. Together, our data reveal a novel regulator of glucan synthases and glucanases, Smi1, which is more important for Bgs4 trafficking, stability, and localization during cytokinesis.
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Pared Celular/metabolismo , Glucosiltransferasas/metabolismo , Proteínas de Schizosaccharomyces pombe/metabolismo , Citoesqueleto de Actina/metabolismo , Actomiosina/metabolismo , Membrana Celular/metabolismo , Pared Celular/fisiología , Citocinesis/fisiología , Glucosiltransferasas/fisiología , Proteínas de la Membrana/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Schizosaccharomyces/metabolismo , Proteínas de Schizosaccharomyces pombe/genética , Proteínas de Schizosaccharomyces pombe/fisiología , Factores de Transcripción/metabolismo , beta-Glucanos/metabolismoRESUMEN
Mechanical properties, size and geometry of cells, and internal turgor pressure greatly influence cell morphogenesis. Computational models of cell growth require values for wall elastic modulus and turgor pressure, but very few experiments have been designed to validate the results using measurements that deform the entire thickness of the cell wall. New wall material is synthesized at the inner surface of the cell such that full-thickness deformations are needed to quantify relevant changes associated with cell development. Here, we present an integrated, experimental-computational approach to analyze quantitatively the variation of elastic bending behavior in the primary cell wall of living Arabidopsis (Arabidopsis thaliana) pavement cells and to measure turgor pressure within cells under different osmotic conditions. This approach used laser scanning confocal microscopy to measure the 3D geometry of single pavement cells and indentation experiments to probe the local mechanical responses across the periclinal wall. The experimental results were matched iteratively using a finite element model of the experiment to determine the local mechanical properties and turgor pressure. The resulting modulus distribution along the periclinal wall was nonuniform across the leaf cells studied. These results were consistent with the characteristics of plant cell walls which have a heterogeneous organization. The results and model allowed the magnitude and orientation of cell wall stress to be predicted quantitatively. The methods also serve as a reference for future work to analyze the morphogenetic behaviors of plant cells in terms of the heterogeneity and anisotropy of cell walls.