Outer Cell Wall Research Articles

Host Recognition for Survival of Plant Pathogens

The intact, healthy plant is a community of cells built in a fortress-like fashion. Plant cells consist of cell wall contains the nucleus and various organelles and all the substances for which the pathogens attack them. The cytoplasm and the organelles it contains are separated from each other by membranes that carry various types of proteins embedded in them (Fig. 5-2). The plant surfaces that come in contact with the environment either consist of cellulose, as in the epidermal cells of roots and in the intercellular spaces of leaf parenchyma cells, or consist of a cuticle that covers the epidermal cell walls, as is the case in the aerial parts of plants. Often an additional layer, consisting of waxes, is deposited outside the cuticle, especially on younger parts of plants Pathogens attack plants because during their evolutionary development they have acquired the ability to live off the substances manufactured by the host plants, and some of the pathogens depend on these substances for survival. Many substances are contained in the protoplast of the plant cells, however, and if pathogens are to gain access to them they must first penetrate the outer barriers formed by the cuticle and/or cell walls. Even after the outer cell wall has been penetrated, further invasion of the plant by the pathogen necessitates the penetration of more cell walls. Furthermore, the plant cell contents are not always found in forms immediately utilizable by the pathogen and must be broken down to units that the pathogen can absorb and assimilate. Moreover, the plant, reacting to the presence and activities of the pathogen, produces structures and chemical substances that interfere with the advance or the existence of the pathogen; if the pathogen is to survive and to continue living off the plant, it must be able to overcome such obstacles. Therefore, for a pathogen to infect a plant it must be able to make its way into and through the plant, obtain nutrients from the plant, and neutral

Genomic characterization and comparative genomics of Chlorella sp. CH2018 from Musi River water, India

Microalgae, a diverse class of photosynthetic eukaryote, can provide food and energy sustainably. Selecting productive strains is crucial for commercial viability. Here, we isolated an axenic microalgal species from Musi River water, and based on its morphology, molecular makeup, and cell wall composition identified it as Chlorella sp. CH2018. Scanning electron microscopy was used to measure the size of cells, which were found to be 3-, 4-, or 5-μm in diameter with a discernible thick outer cell wall. The characterization of algal genomes is imperative for comprehending species, studying metabolic pathways, and modifying genetics. In this study, we sequenced the entire genome of Chlorella sp. CH2018, yielded genome size of 56.83 Mb with 11,143 functionally annotated protein-coding gene models. The Gene Ontology (GO) analysis revealed that Chlorella's predominant metabolic pathway is carbohydrate metabolism. Ortholog comparative analysis of species of phylum Chlorophyta with Chlorella sp. CH2018 showed that the isolated species possesses unique protein families with a maximum number of 6292 ortholog groups with Chlorella sorokiniana. The phylogenetic tree created by concatenating single-copy ortholog sequences demonstrates the uniqueness of Chlorella sp. CH2018, and its genome sequence serves as a genetic resource for future research.

Identification of Kibdelomycin and Related Biosynthetic Gene Clusters and Characterization of the C‐Branching of Amycolose

AbstractMany bacterial natural products contain C‐branched sugars, including components from the outer cell wall or antibiotically active metabolites. The enzymatic C‐branching of keto sugars leading to longer side chains (≥C2) is catalyzed by thiamine diphosphate (ThDP)‐dependent enzymes. Chiral tertiary α‐hydroxy ketones are formed in this process. The ThDP‐dependent enzymes that catalyze C‐branching reactions belong to one of three enzymatic superfamilies: decarboxylases, transketolases, and α‐ketoacid dehydrogenases 2, but branching of keto sugars has only been demonstrated for decarboxylases. In this study, we showed that an α‐ketoacid dehydrogenase is responsible for C‐branching of the deoxyketo sugar amycolose in the biosynthesis of kibdelomycin in Kibdelosporangium sp. MA7385. In addition, we characterized an amino transferase in the same biosynthetic gene cluster (BGC) that accepts a sterically demanding tertiary α‐hydroxy ketone in a downstream reaction. Subsequently, we identified approximately 400 similar BGCs in silico, suggesting that there is a large diversity of possible ThDP‐dependent enzymes catalyzing the C‐branching of keto sugars and subsequent modifications.

Engineering the cell wall reactive groups of Plant Growth Promoting Rhizobacteria by culture strategy for heavy metal removal

This research delved into the effects of nutrient limitation on the level of sporulation and the cadmium adsorption capacity of the bacterium Bacillus sp. isolated from the rhizosphere of endemic soils in the Region of Valparaiso, Chile. The bacteria were subjected to nitrogen limitation in fed-batch mode and were compared to bacteria grown in batch culture without nutrient limitation. The cultures were carried out in a 3 L bioreactor with an external nitrogen supply of ammonium at a flow of 0.123 L h−1. The specific maximum growth rate was 0.42 h−1 in batch and 0.45 h−1 in the exponential phase of the fed-batch.The analysis of sporulation did not show any significant difference between the biomass coming from the fed-batch and batch cultures.It was found that maximum cadmium adsorption capacity varied with culture strategy. The dry biomass grown without nutrient limitation exhibited a maximum adsorption capacity for cadmium of 65.0 mgCd g−1biomass. Conversely, the limited biomass achieved a lower cadmium adsorption capacity of approximately 36.0 mgCd g−1biomass. FTIR analysis showed that nitrogen limitation induced changes in the composition of the outer cell wall, specifically an increase of deacetlylated polysaccharides, reducing the relative amount of secondary amines and proteins from the peptidoglycan matrix. Amino groups from acetylated polysaccharides and proteins have been associated elsewhere with greater cadmium affinity, which could explain the poor results obtained with the nitrogen-restricted biomass. This study shows that new physiological states displaying different adsorption capabilities were effectively obtained by engineering the cell coverage of the bacteria using varying culture strategies. The fed-batch culture proved to be a valuable tool for studying PGPR strains for biosorption and other applications. Exploring diverse nutrient limitations and other pollutants in this bacterium and other members of the PGPR family offer great opportunities to tailor biosorption strategies based on specific conditions, ultimately contributing to sustainable environmental solutions.

The structural organization of the outer tissues in the gametophytic stem of the umbrella moss Hypnodendron menziesii optimizes load bearing.

The ultrastructural design and biochemical organization of the significantly thickened outer tissues of the gametophytic stem of Hypnodendron menziesii optimizes load bearing of the stem. Hypnodendron menziesii is a bryoid umbrella moss growing in high humid conditions on the forest floors of New Zealand. The erect gametophyte bears up to eight whorls of branches in succession, spreading across the stem that bears the heavy weight of branches with highly hydrated leaves. Our investigation using a combination of light microscopy, transmission electron microscopy (TEM), scanning electron microscopy (SEM), and TEM-immunolabeling techniques provided novel information on the structural design and biochemical organization of greatly thickened cell walls of epidermal, hypodermal, and outermost cortical tissues, comparing underlying thin-walled cortical tissues in the gametophytic stem. Probing into the ultrastructure of the cell wall architecture of these target tissues by TEM and SEM revealed the cell walls to display a multilamellar organization, in addition to demonstrating the presence of an electron-dense substance in the cell wall, presumably flavonoids. The pattern of distribution and concentration of rhamnogalacturonan, homogalacturonan, and heteromannan, as determined by immunogold labeling, suggests that it is the combination of structural and molecular design of the cell wall that may optimize the mechanical function of the epidermal, hypodermal, and outer cortical tissues. Statistical relationships between the overall thickness of epidermal, hypodermal, and outer cortical cell walls, the lumen area of cells and the percentage area of cell wall occupied in these tissues at different heights of the stem, and thickness of secondary cell wall layers (L1-L4/5) were explored. The results of these analyses unequivocally support the contribution of outer tissues to the mechanical strength of the resilient stem.

Deletion of the polyketide synthase-encoding gene pks1 prevents melanization in the extremophilic fungus Cryomyces antarcticus.

Cryomyces antarcticus, a melanized cryptoendolithic fungus endemic to Antarctica, can tolerate environmental conditions as severe as those in space. Particularly, its ability to withstand ionizing radiation has been attributed to the presence of thick and highly melanized cell walls, which-according to a previous investigation-may contain both 1,8-dihydroxynaphthalene (DHN) and L-3,4 dihydroxyphenylalanine (L-DOPA) melanin. The genes putatively involved in the synthesis of DHN melanin were identified in the genome of C. antarcticus. Most important is capks1 encoding a non-reducing polyketide synthase (PKS) and being the ortholog of the functionally characterized kppks1 from the rock-inhabiting fungus Knufia petricola. The co-expression of CaPKS1 or KpPKS1 with a 4'-phosphopantetheinyl transferase in Saccharomyces cerevisiae resulted in the formation of a yellowish pigment, suggesting that CaPKS1 is the enzyme providing the precursor for DHN melanin. To dissect the composition and function of the melanin layer in the outer cell wall of C. antarcticus, non-melanized mutants were generated by CRISPR/Cas9-mediated genome editing. Notwithstanding its slow growth (up to months), three independent non-melanized Δcapks1 mutants were obtained. The mutants exhibited growth similar to the wild type and a light pinkish pigmentation, which is presumably due to carotenoids. Interestingly, visible light had an adverse effect on growth of both melanized wild-type and non-melanized Δcapks1 strains. Further evidence that light can pass the melanized cell walls derives from a mutant expressing a H2B-GFP fusion protein, which can be detected by fluorescence microscopy. In conclusion, the study reports on the first genetic manipulation of C. antarcticus, resulting in non-melanized mutants and demonstrating that the melanin is rather of the DHN type. These mutants will allow to elucidate the relevance of melanization for surviving extreme conditions found in the natural habitat as well as in space.

Residual Membrane Fluidity in Mycobacterial Cell Envelope Layers under Extreme Conditions Underlines Membrane-Centric Adaptation.

One of the routes for adaptation to extreme environments is via remodeling of cell membrane structure, composition, and biophysical properties rendering a functional membrane. Collective studies suggest some form of membrane feedback in mycobacterial species that harbor complex lipids within the outer and inner cell wall layers. Here, we study the homeostatic membrane landscape of mycobacteria in response to high hydrostatic pressure and temperature triggers using high pressure fluorescence, mass and infrared spectroscopies, NMR, SAXS, and molecular dynamics simulations. Our findings reveal that mycobacterial membrane possesses unique and lipid-specific pressure-induced signatures that attenuate progression to highly ordered phases. Both inner and outer membrane layers exhibit phase coexistence of nearly identical lipid phases keeping residual fluidity over a wide range of temperature and pressure, but with different sensitivities. Lipidomic analysis of bacteria grown under pressure revealed lipidome remodeling in terms of chain length, unsaturation, and specific long-chained characteristic mycobacterial lipids, rendering a fluid bacterial membrane. These findings could help understand how bacteria may adapt to a broad spectrum of harsh environments by modulating their lipidome to select lipids that enable the maintenance of a fluid functional cell envelope.

Identification of Kibdelomycin and Related Biosynthetic Gene Clusters and Characterization of the C-Branching of Amycolose.

Many bacterial natural products contain C-branched sugars, including components from the outer cell wall or antibiotically active metabolites. The enzymatic C-branching of keto sugars leading to longer side chains (≥C2) is catalyzed by thiamine diphosphate (ThDP)-dependent enzymes. Chiral tertiary α-hydroxy ketones are formed in this process. The ThDP-dependent enzymes that catalyze C-branching reactions belong to one of three enzymatic superfamilies: decarboxylases, transketolases, and α-ketoacid dehydrogenases 2, but branching of keto sugars has only been demonstrated for decarboxylases. In this study, we showed that an α-ketoacid dehydrogenase is responsible for C-branching of the deoxyketo sugar amycolose in the biosynthesis of kibdelomycin in Kibdelosporangium sp. MA7385. In addition, we characterized an amino transferase in the same biosynthetic gene cluster (BGC) that accepts a sterically demanding tertiary α-hydroxy ketone in a downstream reaction. Subsequently, we identified approximately 400 similar BGCs in silico, suggesting that there is a large diversity of possible ThDP-dependent enzymes catalyzing the C-branching of keto sugars and subsequent modifications.

The Complete Assessment of Small Molecule and Peptidomimetic Inhibitors of Sortase A Towards Antivirulence Treatment.

This review covers the most recent advances in the development of inhibitors for the bacterial enzyme sortase A (SrtA). Sortase A (SrtA) is a critical virulence factor, present ubiquitously in Gram-positive bacteria of which many are pathogenic. Sortases are key enzymes regulating bacterial adherence to host cells, by anchoring extracellular matrix-binding proteins to the bacterial outer cell wall. By targeting virulence factors, effective treatment can be achieved, without inducing antibiotic resistance to the treatment. This is a potentially more sustainable, long-term approach to treating bacterial infections, including ones that display multiple resistance to current therapeutics. There are many promising approaches available for SrtA inhibition, some of which have the potential to advance into further clinical development, with peptidomimetic and in vivo active small molecules being among the most promising. There are currently no approved drugs on the market targeting SrtA, despite its promise, adding to the relevance of this review article, as it extends to the pharmaceutical industry additionally to academic researchers.

A specific innate immune response silences the virulence of Pseudomonas aeruginosa in a latent infection model in the Drosophila melanogaster host.

Microbial pathogenicity often depends on the route of infection. For instance, P. aeruginosa or S. marcescens cause acute systemic infections when low numbers of bacteria are injected into D. melanogaster flies whereas flies succumb much slower to the continuous ingestion of these pathogens, even though both manage to escape from the gut compartment and reach the hemocoel. Here, we have developed a latent P. aeruginosa infection model by feeding flies on the bacteria for a short period. The bacteria stably colonize internal tissues yet hardly cause any damage since latently-infected flies live almost as long as noninfected control flies. The apparently dormant bacteria display particular characteristics in terms of bacterial colony morphology, composition of the outer cell wall, and motility. The virulence of these bacteria can however be reactivated upon wounding the host. We show that melanization but not the cellular or the systemic humoral response is the predominant host defense that establishes latency and may coerce the bacteria to a dormant state. In addition, the lasting activation of the melanization responses in latently-infected flies provides a degree of protection to the host against a secondary fungal infection. Latent infection by an ingested pathogen protects against a variety of homologous or heterologous systemic secondary infectious challenges, a situation previously described for the endosymbiotic Wolbachia bacteria, a guard against viral infections.

Impact of secreted glucanases upon the cell surface and fitness of Candida albicans during colonisation and infection

Host recognition of the pathogen-associated molecular pattern (PAMP), β-1,3-glucan, plays a major role in antifungal immunity. β-1,3-glucan is an essential component of the inner cell wall of the opportunistic pathogen Candida albicans. Most β-1,3-glucan is shielded by the outer cell wall layer of mannan fibrils, but some can become exposed at the cell surface. In response to host signals such as lactate, C. albicans shaves the exposed β-1,3-glucan from its cell surface, thereby reducing the ability of innate immune cells to recognise and kill the fungus. We have used sets of barcoded xog1 and eng1 mutants to compare the impacts of the secreted β-glucanases Xog1 and Eng1 upon C. albicans in vitro and in vivo. Flow cytometry of Fc-dectin-1-stained strains revealed that Eng1 plays the greater role in lactate-induced β-1,3-glucan masking. Transmission electron microscopy and stress assays showed that neither Eng1 nor Xog1 are essential for cell wall maintenance, but the inactivation of either enzyme compromised fungal adhesion to gut and vaginal epithelial cells. Competitive barcode sequencing suggested that neither Eng1 nor Xog1 strongly influence C. albicans fitness during systemic infection or vaginal colonisation in mice. However, the deletion of XOG1 enhanced C. albicans fitness during gut colonisation. We conclude that both Eng1 and Xog1 exert subtle effects on the C. albicans cell surface that influence fungal adhesion to host cells and that affect fungal colonisation in certain host niches.

Cutinsomes of Malus Mill. (Rosaceae) leaf and pericarp: genesis, localization, and transport

New data were obtained on specific bionanostructures, cutinsomes, which are involved in the formation of cuticles on the surface of leaf blades and pericarp of Malus domestica Borkh (Malus Mill., Rosaceae)introduced to the mountains at the altitudes of 1200 and 1700 m above sea level. Cutinsomes, which are electron-dense structures of spherical shape, have been identified by transmission electron microscopy. It was demonstrated that plastids can be involved in the synthesis of their constituent nanocomponents. The greatest number of nanoparticles was observed in the granal thylakoid lumen of the chloroplasts in palisade mesophyll cells and pericarp hypodermal cells. The transmembrane transport of cutinsomes into the cell wall cuticle proper by exocytosis has been visualized for the first time. The plasma membrane is directly involved in the excretion of nanostructures from the cell. Nanoparticles of cutinsomes in the form of necklace-like formations line up in a chain near cell walls, merge into larger conglomerates and are loaded into plasmalemma invaginations, and then, in membrane packing, they move into the cuticle, which covers both outer and inner cell walls of external tissues. The original materials obtained by us supplement the ideas about the non-enzymatic synthesis of cuticle components available in the literature and expand the cell compartment geography involved in this process.

Seed morphology in the genus Matthiola R. Br. (Brassicaceae): Identification of species and systematic implications

AbstractSeed coat morphology of 34 species of Matthiola R.Br. has been studied using light and scanning electron microscopy to assess their investigative value for systematic studies. Macro‐ and micromorphological characters, including seed shape, color, size, wing, and radicle position relative to cotyledons, epidermal cell shape, anticlinal boundaries, and periclinal cell wall are presented. Seed shape, color, and radicle position relative to cotyledons have limited taxonomic significance, as their variation is unhelpful. There against, seed size, wing, and seed coat ornamentation present informative characteristics that can be used professionally in distinguishing the studied taxa. Six different types of seed anticlinal cell wall boundaries are recognized and five different outer periclinal cell wall shapes are described. The sculpture of the secondary cell wall varies from striate, micropapillate to microreticulate, and smooth to fine folds. The taxonomic and phylogenetic implications of seed coat micromorphology were compared with those of the existing gross morphological and molecular data. Seed character analysis offered useful data for evaluating the taxonomy of Matthiola both on subgeneric and sectional levels. Seed micromorphological characters provide evidence that M. odoratissima and M. ovatifolia should be preserved as separate species. In addition, M. ghorana is not conspecific with M. graminea. Furthermore, M. tricuspidata was unique in the seed characters. A key to the identification of the examined taxa based on seed characters is provided.

Anatomical limitations in adventitious root formation revealed by magnetic resonance imaging, infrared spectroscopy, and histology of rose genotypes with contrasting rooting phenotypes.

Adventitious root (AR) formation is one of the most important developmental processes in vegetative propagation. Although genotypic differences in rose rooting ability are well known, the causal factors are not well understood. The rooting of two contrasting genotypes, 'Herzogin Friederike' and 'Mariatheresia', was compared following a multiscale approach. Using magnetic resonance imaging, we non-invasively monitored the inner structure of stem cuttings during initiation and progression of AR formation for the first time. Spatially resolved Fourier-transform infrared spectroscopy characterized the chemical composition of the tissues involved in AR formation. The results were validated through light microscopy and complemented by immunolabelling. The outcome demonstrated similarity of both genotypes in root primordia formation, which did not result in root protrusion through the shoot cortex in the difficult-to-root genotype 'Mariatheresia'. The biochemical composition of the contrasting genotypes highlighted main differences in cell wall-associated components. Further spectroscopic analysis of 15 contrasting rose genotypes confirmed the biochemical differences between easy- and difficult-to-root groups. Collectively, our data indicate that it is not the lack of root primordia limiting AR formation in these rose genotypes, but the firmness of the outer stem tissue and/or cell wall modifications that pose a mechanical barrier and prevent root extension and protrusion.

Equisetum arvense as a silica fertilizer

The aim was to use the agricultural weed and silica (Si) hyperaccumulator Equisetum arvense as Si fertilizer in plant cultivation. We investigated (1) the Si uptake in various Equisetum species, (2) where Si accumulates in the Equisetum plant, (3) processing methods to release as much Si as possible from dried, ground E. arvense plants and (4) which treatment yields gives the highest uptake of Si in young wheat plants cultivated in soil containing ground E. arvense. The results showed that E. arvense containes 22% Si and was among the best Si accumulators. Equisetum arvense accumulates Si as both soluble and firmly bound fractions. Amorphous silica (SiO2) accumulates in the outer cell walls of epidermis of the entire plant. Regarding the processing method, a longer treatment time, greater concentration of Equisetum, boiling, and the addition of sodium bicarbonate increased the Si availability in ground, dried E. arvense. The addition of untreated, ground, dried E. arvense to the soil, corresponding to 160 kg Si ha−1, increased the available Si in the soil and the Si uptake in wheat plants by five-fold, compared with the control. Boiling the ground E. arvense increased the Si uptake by 10 times, and the of sodium bicarbonate increased the availability and uptake by 40 times, compared with the control. In conclusion, dried, ground E. arvense can be used as a Si fertilizer as is, after boiling for a slightly better effect, or with sodium bicarbonate (up to a similar amount as the ground material) for best effect.

Hemocompatible polyethylenimine-grafted poly (glycidyl methacrylate) based microspheres for effective endotoxin removal

Endotoxin (also known as lipopolysaccharide, LPS), the major component in the outer cell wall of Gram-negative bacteria, is considered to be a main cause of sepsis. To reduce the mortality of sepsis, adsorbents with positively charged surfaces like polyethylenimine (PEI) are commonly employed to adsorb endotoxins; however, there are hidden dangers in binding stability and hemocompatibility of PEI-grafted materials. Herein, poly (glycidyl methacrylate) microspheres prepared by electro-spraying are chosen as the substrate, the surface epoxy groups are utilized to chemically immobilize PEI molecules. Then, heparin coatings are introduced through electrostatic interaction to obtain composite microspheres as hemoperfusion adsorbent. As a result, the hemocompatibility of the microspheres is significantly improved after the introduction of the heparin coatings. Compared to the microspheres only grafting PEI, the hemolysis ratio has a huge decrease from 4.6% to 1.0%. The adsorption process of the composite microspheres is more consistent with Sips isotherm model with a theoretical maximum endotoxin adsorption capacity of 2659.1 EU/g in normal saline solution. And the microspheres exhibit high endotoxin clearance ratio of 81.3% in static plasma (C0 = 12.6 EU/mL, exceeding critical illness level). In vitro dynamic adsorption experiments also indicate that the microspheres remove 86.6% of endotoxin in septic plasma (C0 = 7.2 EU/mL, exceeding critical illness level) within 1 h. We hope that the composite microspheres could have good application potential as an efficiency endotoxin adsorbent in sepsis treatment.

High-Performance Engineered Composites Biofabrication Using Fungi.

Various natural polymers offer sustainable alternatives to petroleum-based adhesives, enabling the creation of high-performance engineered materials. However, additional chemical modifications and complicated manufacturing procedures remain unavoidable. Here, a sustainable high-performance engineered composite that benefits from bonding strategies with multiple energy dissipation mechanisms dominated by chemical adhesion and mechanical interlocking is demonstrated via the fungal smart creative platform. Chemical adhesion is predominantly facilitated by the extracellular polymeric substrates and glycosylated proteins present in the fungal outer cell walls. The dynamic feature of non-covalent interactions represented by hydrogen bonding endows the composite with extensive unique properties including healing, recyclability, and scalable manufacturing. Mechanical interlocking involves multiple mycelial networks (elastic modulus of 2.8GPa) binding substrates, and the fungal inner wall skeleton composed of chitin and β-glucan imparts product stability. The physicochemical properties of composite (modulus of elasticity of 1455.3MPa, internal bond strength of 0.55MPa, hardness of 82.8, and contact angle of 110.2°) are comparable or even superior to those of engineered lignocellulosic materials created using petroleum-based polymers or bioadhesives. High-performance composite biofabrication using fungi may inspire the creation of other sustainable engineered materials with the assistance of the extraordinary capabilities of living organisms.

The Metallotolerance and Biosorption of As(V) and Cr(VI) by Black Fungi.

A collection of 34 melanized fungi isolated previously from anthropogenic contaminated sites were assessed for their tolerance to toxic concentrations of As(V) and Cr(VI) anions. Three strains of the species Cyphellophora olivacea, Rhinocladiella similis, and Exophiala mesophila (Chaetothyriales) were identified as hyper-metallotolerant, with estimated IC50 values that ranged from 11.2 to 16.9 g L-1 for As(V) and from 2.0 to 3.4 g L-1 for Cr(VI). E. mesophila and R. similis were selected for subsequent assays on their biosorption capacity and kinetics under different pH values (4.0 and 6.5) and types of biomass (active and dead cells and melanin extracts). The fungal biosorption of As(V) was relatively ineffective, but significant removal of Cr(VI) was observed from liquid cultures. The Langmuir model with second-order kinetics showed maximum sorption capacities of 39.81 mg Cr6+ g-1 for R. similis and 95.26 mg Cr6+ g-1 for E. mesophila on a dry matter basis, respectively, while the kinetic constant for these two fungi was 1.32 × 10-6 and 1.39 × 10-7 g (mg Cr6+ min)-1. Similar experiments with melanin extracts of E. mesophila showed maximum sorption capacities of 544.84 mg Cr6+ g-1 and a kinetic constant of 1.67 × 10-6 g (mg Cr6+ min)-1. These results were compared to bibliographic data, suggesting that metallotolerance in black fungi might be the result of an outer cell-wall barrier to reduce the diffusion of toxic metals into the cytoplasm, as well as the inner cell wall biosorption of leaked metals by melanin.

The nonlinear mechanics of highly extensible plant epidermal cell walls.

Plant epidermal cell walls maintain the mechanical integrity of plants and restrict organ growth. Mechanical analyses can give insights into wall structure and are inputs for mechanobiology models of plant growth. To better understand the intrinsic mechanics of epidermal cell walls and how they may accommodate large deformations during growth, we analyzed a geometrically simple material, onion epidermal strips consisting of only the outer (periclinal) cell wall, ~7 μm thick. With uniaxial stretching by >40%, the wall showed complex three-phase stress-strain responses while cyclic stretching revealed reversible and irreversible deformations and elastic hysteresis. Stretching at varying strain rates and temperatures indicated the wall behaved more like a network of flexible cellulose fibers capable of sliding than a viscoelastic composite with pectin viscosity. We developed an analytic framework to quantify nonlinear wall mechanics in terms of stiffness, deformation, and energy dissipation, finding that the wall stretches by combined elastic and plastic deformation without compromising its stiffness. We also analyzed mechanical changes in slightly dehydrated walls. Their extension became stiffer and more irreversible, highlighting the influence of water on cellulose stiffness and sliding. This study offers insights into the structure and deformation modes of primary cell walls and presents a framework that is also applicable to tissues and whole organs.

Rapid and high accurate identification of Escherichia coli active and inactivated state by hyperspectral microscope imaging combing with machine learning algorithm

Rapid identification of the active state of foodborne bacteria is crucial for ensuring the safety and quality control of food or pharmaceutical products. In this study, a combination of hyperspectral microscope imaging (HMI) and machine learning algorithm is employed for the identification of active state of Escherichia coli (E. coli). Hyperspectral microscope images of live, 100 ℃ heat inactivation and 121 ℃ high-pressure inactivation of E. coli are collected in wavelength range of 370–1060 nm. Savitzky-Golay (SG) smoothing combing with normalization is used for spectra preprocessing. And principal component analysis (PCA) is employed for spectral dimension reduction. Four different regions of interest (ROIs), including the entire bacterial cell ROI (cell), the outer cell wall ROI (cell_r), the membrane structure ROI (cell_w) formed by the cell wall and cell membrane, and the central of the cell ROI (cell_cy), are extracted and used as model input variables to investigate the influence on the modeling results. Five model algorithms, support vector machines (SVM), random forests (RF), k-nearest neighbors (KNN) algorithms, discriminant analysis (DA) classifiers, and long short-term memory (LSTM) neural networks are used and compared. Modeling results with spectral data of cell_r perform better than those with other ROIs. Accuracy of the models with data of the cell_r ROI are as follows: 79.78% for SVM, 95.11% for RF, 91.33% for KNN, 98.22% for DA, and 93.78% for LSTM. DA achieves the highest classification accuracy. The results show that high-temperature inactivation induces changes in bacterial tissue and morphology, resulting in certain spectral differences among bacteria in three different states. The combination of hyperspectral microscope imaging and machine learning algorithm can provide an effective method for identification of active and inactive states of E. coli. Furthermore, the model, constructed with the data of cell_r ROI, exhibits the best performance in identification.