Cell Wall Morphology Research Articles

An Analysis of the Mechanism About CO2 Enrichment Promoting Carbohydrate Metabolism in Cucumber (Cucumis sativus L.) Leaves.

Elevated CO2 can affect the synthesis and distribution of photosynthetic assimilates. However, the carbohydrate metabolism molecular mechanism of cucumber leaves in response to CO2 enrichment is unclear. Therefore, it is of great significance to investigate the key functional regulatory genes in cucumber. In this study, the growth of cucumber leaves under different CO2 conditions was compared. The results showed that under CO2 enrichment, leaf area increased, the number of mesophyll cells increased, stomata enlarged, and more starch grains accumulated in the chloroplasts. Compared with the control, the starch and soluble sugar content of leaves were maximally increased by 194.1% and 55.94%, respectively; the activities of fructose-1,6-bisphosphatase (FBPase), ADPG pyrophosphorylase (AGPase), starch synthase (SSS), sucrose phosphate synthase (SPS), sucrose synthase (SS) and invertase (Inv) were maximally increased by 36.91%, 66.13%, 33.18%, 21.7%, 54.11%, and 46.01%, respectively. Through transcriptome analysis, a total of 1,582 differential expressed genes (DEGs) were identified, in which the starch and sucrose metabolism pathway was significantly enriched, and 23 genes of carbon metabolism were screened. Through metabolome analysis, a total of 22 differential accumulation metabolites (DAMs) were identified. Moreover, D-glucose and D(+)-glucose were significantly accumulated, showing upregulation 2.4-fold and 2.6-fold, respectively. Through combined analysis of transcriptome and metabolome, it was revealed that seven genes were highly related to D-glucose, and Csa6G153460 (AGPase), Csa5G612840 (β-glucosidase), and Csa4G420150 (4-α-glucanotransferase) were significantly correlated to the carbohydrate regulatory network. Furthermore, the mechanism of CO2 enrichment that promotes carbohydrate metabolism in leaves at the molecular level was revealed. This mechanism advances the development of the cell wall and leaf morphology by activating the expression of key genes and improving enzyme activity.

Endophytic Fungus UJ3-2 from Urtica fissa: Antibacterial Activity and Mechanism of Action against Staphylococcus aureus

Taking the endophytic fungus UJ3-2, isolated from Urtica fissa, as the experimental material, this study aimed to explore the composition of its metabolites and the underlying mechanisms by which it inhibits Staphylococcus aureus. Initially, the MIC, MBC, inhibitory curves, biofilm growth, and extracellular nucleic acids and proteins of S. aureus in response to the metabolites were measured. Secondly, PI staining and SEM were used to evaluate the impact of the metabolites on the integrity of the cell wall and overall morphology of S. aureus. Additionally, UPLC-MS was employed to analyze the composition of the secondary metabolites. The UJ3-2 strain was identified as Xylaria grammica based on ITS sequencing and designated as Xylaria grammica UJ3-2. Our results revealed that the metabolites of UJ3-2 exhibited excellent in vitro antibacterial activity against S. aureus, with both MIC and MBC values of 3.125 mg/mL. The inhibitory curve confirmed that 1 MIC of UJ3-2 metabolites could completely inhibit the growth of S. aureus within 24 h. With increasing concentrations of UJ3-2 metabolites, the growth of S. aureus biofilms was significantly suppressed, and obvious leakage of nucleic acids and proteins was observed. PI fluorescence staining indicated that various concentrations of UJ3-2 metabolites disrupted the integrity of the S. aureus cell membrane. SEM observation revealed that the treated S. aureus surfaces became rough, and the bacteria shrank and adhered to each other, showing a dose-dependent effect. UPLC-MS analysis suggested that the main components of the fermented metabolites were 6-oxocineole (17.92%), (S)-2-acetolactate (9.91%), 3-methyl-cis,cis-muconate (4.36%), and 8-oxogeranial (3.17%). This study demonstrates that the endophytic fungus UJ3-2 exhibits remarkable in vitro antibacterial effects against S. aureus, primarily by enhancing the permeability of the S. aureus cell membrane, causing the leakage of its intracellular contents, and altering the bacterial surface morphology to inhibit the pathogen. The endophytic fungus UJ3-2 has a good antibacterial effect on S. aureus, which gives it certain application prospects in the screening and industrial production of new and efficient natural antibacterial active substances.

Phytochemical and Functional Diversity of Enzyme-Assisted Extracts from Hippophae rhamnoides L., Aralia cordata Thunb., and Cannabis sativa L.

Plant leaves are a source of essential phenolic compounds, which have numerous health benefits and can be used in multiple applications. While various techniques are available for recovering bioactive compounds from by-products, more data are needed on enzyme-assisted extraction (EAE). The aim of this study was to compare EAE and solid-liquid extraction (SLE), to evaluate the impact on bioactive compounds' extraction yield, phytochemical composition, and the antioxidant, antimicrobial, and antidiabetic properties of Aralia cordata leaves and roots, sea buckthorn Hippophae rhamnoides, and hemp Cannabis sativa leaves. The results indicate that EAE with Viscozyme L enzyme (EAE_Visc) extracts of the tested plant leaves possess the highest yield, antioxidant activity, and total phenolic content. Moreover, the EAE_Visc extract increased by 40% the total sugar content compared to the control extract of A. cordata root. Interestingly, the sea buckthorn leaf extracts exhibited α-glucosidase inhibitory activity, which reached an almost 99% inhibition in all extracts. Furthermore, the sea buckthorn leaves SLE and EAE_Visc extracts possess antibacterial activity against Staphylococcus aureus. Additionally, scanning electron microscopy was used to examine changes in cell wall morphology after EAE. Overall, this study shows that EAE can be a promising method for increasing the yield and improving the functional properties of the resulting extracts in a fast and sustainable way compared to SLE.

Investigation of heat-resistant antifungal agents from Bacillus amyloliquefaciens and Bacillus subtilis for biocontrol of mycotoxigenic fungi

Fungal infections of food and feed with toxigenic strains results in the accumulation of harmful metabolites. In this study, the antifungal activity of two indigenous bacterial strains Bacillus amyloliquefaciens and Bacillus subtilis isolated from camel feed was investigated. The potential of the bacterial cell free supernatant (CFS) to reduce mycotoxins synthesis was evaluated, and the impact of their antifungal metabolites on the fungal hyphae was examined. High antagonistic activity was exhibited by both bacterial strains. The best activity was observed against two mycotoxigenic A. niger strains (Sp31 and Sp33) with inhibition zones of 36.75 ± 0.7 mm and 35.75 ±1.5 mm, by strains B. amyloliquefaciens and B. subtilis, respectively. The effects of these bacterial strains’ CFSs on mycotoxins synthesis were investigated, and both CFSs were found to reduce the mycotoxins synthesis by A. parasiticus, A. niger and Penicillium spp., in a dose-dependent manner. Moreover, the fungal cell wall morphology was significantly altered by the bacterial CFS at a very low concentration of 1 %. The heat stability of the antifungal metabolites was evaluated at various temperatures ranging from −20 °C to 100 °C, and the metabolites were found to be highly stable, retaining their antifungal activity against A. niger. The antifungal activity of B. amyloliquefaciens and B. subtilis was stable at −20 °C for 236 and 34 days of storage, respectively. These findings suggest that both bacterial strains are exceptional candidates in the biocontrol of mycotoxigenic fungi and in the reduction of their toxic metabolites.

Aluminum oxide, cobalt aluminum oxide, and aluminum-doped zinc oxide nanoparticles as an effective antimicrobial agent against pathogens

Since the clock of antimicrobial resistance was set, modern medicine has shed light on a new cornerstone in technology to overcome the worldwide dread of the post-antimicrobial era. Research organizations are exploring the use of nanotechnology to modify metallic crystals from macro to nanoscale size, demonstrating significant interest in the field of antimicrobials. Herein, the antimicrobial activities of aluminum oxide (Al2O3), cobalt aluminum oxide (CoAl2O4), and aluminum doped zinc oxide (Zn0.9Al0.1O) nanoparticles were examined against some nosocomial pathogens. The study confirmed the formation and characterization of Al2O3, CoAl2O4, and Zn0.9Al0.1O nanoparticles using various techniques, revealing the generation of pure nanoscale nanoparticles. With inhibition zones ranging from 9 to 14 mm and minimum inhibitory concentrations varying from 4 mg/mL to 16 mg/mL, the produced nanoparticles showed strong antibacterial activity against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus. Meanwhile, the bactericidal concentrations ranged from 8 mg/mL to 40 mg/mL. In culture, Zn0.9Al0.1O NPs demonstrated a unique ability to inhibit the development of nosocomial infections with high bactericidal activity (8 mg/mL). Transmission electron microscope images revealed changes in cell shape, bacterial cell wall morphology, cytoplasmic membrane, and protoplasm due to the introduction of tested nanoparticles. These results pave the way for the use of these easily bacterial wall-piercing nanoparticles in combination with potent antibiotics to overcome the majority of bacterial strains' resistance.

Novel Aminocoumarin Derivatives against Phytopathogenic Fungi: Design, Synthesis and Structure-Activity Relationships.

Phytopathogenic fungi is the most devastating reason for the decrease of the agricultural production and food safety. To develop new fungicidal agents for resistance concerning, a novel series of aminocoumarin derivatives were synthesized and their fungicidal activity were investigated both in vitro and in vivo. Transmission electron microscope (TEM), scanning electron microscope (SEM), RNA-Seq, 3D-QSAR and molecular docking were applied to reveal the underlying anti-fungal mechanisms. Most of the compounds exhibited significant fungicidal activity. Notably, compound 10c had a more extensive fungicidal effect than positive control. TEM indicated that compound 10c could cause abnormal morphology of cell walls, vacuoles and release of cellular contents. Transcriptional analysis data indicated that 895 and 653 out of 1548 differential expressed genes (DEGs) were up-regulated and down-regulated respectively. The Go and KEGG enrichment indicated that the coumarin derivatives could induce significant changes of succinate dehydrogenase (SDH), Acetyl-coenzyme A synthetase (ACCA) and pyruvate dehydrogenase (PDH) genes, which contributed to the disorders of glucolipid metabolism and the dysfunction of mitochondrial. The results demonstrated that aminocoumarins with schiff-base as core moieties could be the promising lead compounds for the discovery of novel fungicides.

In vitro evaluation of the application of an optimized xylanase cocktail for improved monogastric feed digestibility.

Xylanases from glycoside hydrolase (GH) families 10 and 11 are common feed additives for broiler chicken diets due to their catalytic activity on the nonstarch polysaccharide xylan. This study investigated the potential of an optimized binary GH10 and GH11 xylanase cocktail to mitigate the antinutritional effects of xylan on the digestibility of locally sourced chicken feed. Immunofluorescence visualization of the activity of the xylanase cocktail on xylan in the yellow corn of the feed showed a substantial collapse in the morphology of cell walls. Secondly, the reduction in the viscosity of the digesta of the feed by the cocktail showed an effective degradation of the soluble fraction of xylan. Analysis of the xylan degradation products from broiler feeds by the xylanase cocktail showed that xylotriose and xylopentaose were the major xylooligosaccharides (XOS) produced. In vitro evaluation of the prebiotic potential of these XOS showed that they improved the growth of the beneficial bacteria Streptococcus thermophilus and Lactobacillus bulgaricus. The antibacterial activity of broths from XOS-supplemented probiotic cultures showed a suppressive effect on the growth of the extraintestinal infectious bacterium Klebsiella pneumoniae. Supplementing the xylanase cocktail in cereal animal feeds attenuated xylan's antinutritional effects by reducing digesta viscosity and releasing entrapped nutrients. Furthermore, the production of prebiotic XOS promoted the growth of beneficial bacteria while inhibiting the growth of pathogens. Based on these effects of the xylanase cocktail on the feed, improved growth performance and better feed conversion can potentially be achieved during poultry rearing.

Transforming Wood into a High-Performance Engineering Material via Cellulose Nanocrystal Impregnation

Abstract The demand for wood in construction has led to shortages of strong wood types, causing a shift to costlier alternatives like concrete and nonbiodegradable materials, prompting the investigation of modifying softwoods for better engineering properties. This study investigates the optimization of a multistep impregnation process utilizing functionalized cellulose nanocrystals (f-CNCs) to enhance softwood properties. The process involves alkali delignification, ultrasonication, and vacuum pressure treatment to improve wood porosity and in turn improve CNC impregnation with uniform dispersion. Microstructural analyses through field emission scanning electron microscopy and atomic force microscopy (AFM) offer detailed insights into cell wall morphology and surface topography, whereas Fourier transform infrared spectroscopy highlights compositional shifts resulting from f-CNC impregnation. Mechanical testing demonstrates significant improvements for treated woods, particularly a 67 percent increase in modulus of elasticity for the 2 percent CNC-treated group compared with the control group; a 71 percent increase in modulus of rupture was observed for 2 percent CNC-, 3 percent NaOH-, and 2 percent acetic acid-treated group compared with the control sample. The sample delignified with 3 percent NaOH and impregnated by 2 percent f-CNC emerged as particularly effective. This research sets the stage for potential advancements in strengthening softwood using CNC, including a novel AFM method and alternative impregnation techniques like the Lowry method, inviting further exploration.

Multivalent inhibition of the Aspergillus fumigatus KDNase.

Aspergillus fumigatus is a saprophytic fungus and opportunistic pathogen often causing fatal infections in immunocompromised patients. Recently AfKDNAse, an exoglycosidase hydrolyzing 3-deoxy-D-galacto-D-glycero-nonulosonic acid (KDN), a rare sugar from the sialic acid family, was identified and characterized. The principal function of AfKDNAse is still unclear, but a study suggests a critical role in fungal cell wall morphology and virulence. Potent AfKDNAse inhibitors are required to better probe the enzyme's biological role and as potential antivirulence factors. In this work, we developed a set of AfKDNAse inhibitors based on enzymatically stable thio-KDN motifs. C2, C9-linked heterodi-KDN were designed to fit into unusually close KDN sugar binding pockets in the protein. A polymeric compound with an average of 54 KDN motifs was also designed by click chemistry. Inhibitory assays performed on recombinant AfKDNAse showed a moderate and strong enzymatic inhibition for the two classes of compounds, respectively. The poly-KDN showed more than a nine hundred fold improved inhibitory activity (IC50 = 1.52 ± 0.37 μM, 17-fold in a KDN molar basis) compared to a monovalent KDN reference, and is to our knowledge, the best synthetic inhibitor described for a KDNase. Multivalency appears to be a relevant strategy for the design of potent KDNase inhibitors. Importantly, poly-KDN was shown to strongly decrease filamentation when co-cultured with A. fumigatus at micromolar concentrations, opening interesting perspectives in the development of antivirulence factors.

Medicinal Remedies for the Treatment of Methicillin Resistant Staphylococcus Aureus

Antimicrobial resistance has led to the rise of formidable microbes like methicillin-resistant Staphylococcus aureus (MRSA). This infection can be acquired at the hospital and in a community, making the search for efficient treatment critical, given the rise in inefficacy of conventional treatments such as vancomycin. Phytocompounds can offer protection against these threats. They not only have the ability to kill MRSA but also enhance its susceptibility to previously ineffective antimicrobial agents. Plant-derived anti-MRSA agents can produce therapeutic effects comparable to some conventional treatments, and in some cases, they may even outperform an antibiotic. This article provided many examples of botanical compounds that have proven potent agents against MRSA in both in vitro and in vivo studies. These compounds achieve these effects by inhibiting the resistance mechanisms in MRSA, breaking down its defenses, making it vulnerable, and eventually killing it. Many of these compounds are inherently bactericidal; however, they may produce a synergistic response when combined with allopathic antibiotics. For example, plumbagin, from Plumbago zeylanica, not only disrupts MRSA cell wall morphology, eliciting cytoplasmic changes but also produces a synergistic effect with ciprofloxacin and piperacillin. Moreover, honokiol and magnolol glycosides, from Magnolia officinalis, reversed the quintessential resistance of MRSA by repressing genes, responsible for resistance development. The possibilities are indeed endless, but further research and financial investments are required for proper drug development.

Optimization of Enzyme-Assisted Extraction of Bioactive Compounds from Sea Buckthorn (Hippophae rhamnoides L.) Leaves: Evaluation of Mixed-Culture Fermentation.

Hippophae rhamnoides L. leaves possess a remarkable amount of polyphenols that could serve as a natural remedy in various applications. In comparison, numerous techniques, such as conventional and high-pressure techniques, are available for extracting the bioactive fractions from sea buckthorn leaves (SBL). However, enzyme-assisted extraction (EAE) of SBL has not been comprehensively studied. The aim of this study was to optimize critical EAE parameters of SBL using the cellulolytic enzyme complex, Viscozyme L, to obtain a high-yield extract with a high concentration of bioactive compounds. In order to determine the optimal conditions for EAE, the study employed a central composite design and response surface methodology to analyze the effects of four independent factors (pH, temperature, extraction time, and enzyme concentration) on two different responses. Our findings indicated that under optimal conditions (3:15 h extraction, temperature 45 °C, pH 4.9, and 1% Viscozyme L v/w of leaves DW), EAE yielded 28.90 g/100 g DW of the water-soluble fraction. Furthermore, the EAE-optimized liquid extract was continuously fermented using an ancient fermentation starter, Tibetan kefir grains, which possess lactic acid bacteria (LAB) and have significant potential for use in biopreservation. Interestingly, the results indicated various potential prebiotic characteristics of LAB. Additionally, alterations in the cell wall morphology of the SBL residue after EAE were examined using scanning electron microscopy (SEM). This study significantly optimized EAE parameters for sea buckthorn leaves, providing a promising natural source of bioactive compounds for various applications, such as nutraceuticals, functional foods, and high-value products.

Comparison of cell wall changes of two different types of apple cultivars during fruit development and ripening

Fruit development and ripening is a complex procedure (Malus×domestica Borkh.) and can be caused by various factors such as cell structure, cell wall components, and cell wall hydrolytic enzymes. In our study, we focused on the variations in fruit firmness, cell wall morphology and components, the activity of cell wall hydrolytic enzymes and the expression patterns of associated genes during fruit development in two different types of apple cultivars, the hard-crisp cultivar and the loose-crisp cultivar. In this paper, the aim was to find out the causes of the texture variations between the different type cultivars. Cell wall materials (CWMs), hemicellulose and cellulose content were strongly associated with variations in fruit firmness during the fruit development. The content of water soluble pectin (WSP) and chelator soluble pectin (CSP) gradually increased, while the content of ionic soluble pectin (ISP) showed inconsistent trends in the four cultivars. The activities of polygalacturonase (PG), β-galactosidase (β-gal), cellulase (CEL), and pectate lyase (PL) gradually increased in four cultivars. And the activities of PG, β-gal, and CEL were higher in ‘Fuji’ and ‘Honeycrisp’ fruit with the fruit development, while the activity of PL of ‘Fuji’ and ‘Honeycrisp’ was lower than that of ‘ENVY’ and ‘Modi’. Both four cultivars of fruit cells progressively became bigger as the fruit expanded, with looser cell arrangements and larger cell gaps. According to the qRT-PCR, the relative expression levels of MdACO and Mdβ-gal were notably enhanced. Our study showed that there were large differences in the content of ISP and hemicellulose, the activity of PL and the relative expression of Mdβ-gal between two different types of apple cultivars, and these differences might be responsible for the variations in the texture of the four cultivars.

“Exogenous boron alleviates salt stress in cotton by maintaining cell wall structure and ion homeostasis”

Salt stress is considered one of the major abiotic stresses that impair agricultural production, while boron (B) is indispensable for plant cell composition and has also been found to alleviate salt stress. However, the regulatory mechanism of how B improves salt resistance via cell wall modification remains unknown. The present study primarily focused on investigating the mechanisms of B-mediated alleviation of salt stress in terms of osmotic substances, cell wall structure and components and ion homeostasis. The results showed that salt stress hindered plant biomass and root growth in cotton. Moreover, salt stress disrupted the morphology of the root cell wall as evidenced by Transmission Electron Microscope (TEM) analysis. The presence of B effectively alleviated these adverse effects, promoting the accumulation of proline, soluble protein, and soluble sugar, while reducing the content of Na+ and Cl− and augmenting the content of K+ and Ca2+ in the roots. Furthermore, X-ray diffraction (XRD) analysis demonstrated a decline in the crystallinity of roots cellulose. Boron supply also reduced the contents of chelated pectin and alkali-soluble pectin. Fourier-transform infrared spectroscopy (FTIR) analysis further affirmed that exogenous B led to a decline in cellulose accumulation. In conclusion, B offered a promising strategy for mitigating the adverse impact of salt stress and enhancing plant growth by countering osmotic and ionic stresses and modifying root cell wall components. This study may provide invaluable insights into the role of B in ameliorating the effects of salt stress on plants, which could have implications for sustainable agriculture.

Qualitative and quantitative assessment of diatom deformities and protoplasmic condition under metal and metalloid stress.

Metals and metalloids are toxic, persistent, and non-biodegradable and can be biomagnified (e.g., Hg), and therefore pose a serious threat to the algal flora of aquatic ecosystems. This laboratory study tested the effects of metals (Zn, Fe, and Hg) and a metalloid (As) on the cell wall morphology and protoplasmic content of living cells of six widespread diatom genera over 28days. Diatoms exposed to Zn and Fe had a higher frequency of deformed diatom frustules (> 1%) compared to the As, Hg, and control treatments (< 1%). Deformities in the valve outline and striae were found in all treatments, including the control, whereas deformed raphes and more than one type of deformity were more prevalent under Zn and Hg stress. The order of toxicity is as follows: Zn > Fe > Hg≈As. Deformities were more frequent in Achnanthes and Diploneis (adnate forms) than in the motile genera of Nitzschia and Navicula. The correlation between the % healthy diatoms and % deformities in all six genera showed a negative relationship with the integrity of protoplasmic content (i.e., greater alteration in protoplasmic content was associated with greater frustule deformation). We conclude that diatom deformities can be a good indicator of metal and metalloid stress in waterbodies and are very useful in the rapid biomonitoring of aquatic ecosystems.

Genome-Wide Identification and Expression Profiling of the FORMIN Gene Family Implies Their Potential Functions in Abiotic Stress Tolerance in Rice (Oryza sativa)

FORMIN proteins, which contain FH1 and FH2 domains, play crucial roles in the growth and development of organisms. However, the functions of FORMINs in rice (Oryza sativa) remain largely unclear. In this study, a total of 17 FORMIN genes in rice genome were identified, and their distribution on chromosomes, gene structure, as well as protein structure were investigated. According to their protein structural and phylogenetic features, these 17 rice FORMIN genes were classified into two distinct subfamilies. The results of subcellular localization prediction showed that rice FORMINs were located in cytosol, Golgi complex, endoplasmic reticulum, extracellular, or vacuole. Protein–protein interaction (PPI) prediction results showed that FORMIN proteins might answer hormone signals and be involved in cytoskeleton dynamics regulation and cell wall morphology regulation. The gene expression analysis by using qRT-PCR indicated that a number of rice FORMIN genes were induced by auxin/indole-3-acetic acid (Aux/IAA) and abscisic acid (ABA). Importantly, some of the FORMIN genes also exhibited cadmium (Cd) and drought stress-responding expression patterns, suggesting that FORMIN genes may play roles in rice while dealing with drought or Cd stress. Overall, our research may shed light on the understanding of the biological functions of rice FORMINs.

Effects of Steaming on Sweet Potato Soluble Dietary Fiber: Content, Structure, and Lactobacillus Proliferation In Vitro.

The influence of steaming treatment on the soluble dietary fiber (SDF) of sweet potato was investigated. The SDF content increased from 2.21 to 4.04 g/100 g (in dry basis) during 20 min of steaming. The microcosmic morphology of the fractured cell wall indicated the release of SDF components during steaming. The SDF from fresh (SDF-F) and 20 min steamed (SDF-S) sweet potato was characterized. The neutral carbohydrates and uronic acid levels in SDF-S were significantly higher than SDF-F (59.31% versus 46.83%, and 25.36% versus 9.60%, respectively) (p < 0.05). The molecular weight of SDF-S was smaller than SDF-F (5.32 kDa versus 28.79 kDa). The probiotic property was evaluated by four Lactobacillus spp. fermentation in vitro with these SDF as carbon source, using inulin as the references. SDF-F showed the best proliferation effects on the four Lactobacillus spp. in terms of the OD600 and pH in cultures, and the highest production of propanoic acid and butyric acid after 24 h fermentation. SDF-S presented higher Lactobacillus proliferation effects, but slight lower propanoic acid and butyric acid production than inulin. It was concluded that 20 min of steaming released SDF with inferior probiotic properties, which might derive from the degraded pectin, cell wall components, and resistant dextrin.

Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium.

Obligate intracellular bacteria of the order Rickettsiales include important human pathogens. However, our understanding of the biology of Rickettsia species is limited by challenges imposed by their obligate intracellular lifestyle. To overcome this roadblock, we developed methods to assess cell wall composition, growth, and morphology of Rickettsia parkeri, a human pathogen in the spotted fever group of the Rickettsia genus. Analysis of the cell wall of R. parkeri revealed unique features that distinguish it from free-living alphaproteobacteria. Using a novel fluorescence microscopy approach, we quantified R. parkeri morphology in live host cells and found that the fraction of the population undergoing cell division decreased over the course of infection. We further demonstrated the feasibility of localizing fluorescence fusions, for example, to the cell division protein ZapA, in live R. parkeri for the first time. To evaluate population growth kinetics, we developed an imaging-based assay that improves on the throughput and resolution of other methods. Finally, we applied these tools to quantitatively demonstrate that the actin homologue MreB is required for R. parkeri growth and rod shape. Collectively, a toolkit was developed of high-throughput, quantitative tools to understand growth and morphogenesis of R. parkeri that is translatable to other obligate intracellular bacteria.

Antibacterial Mechanism of Patrinia scabiosaefolia Against Methicillin Resistant Staphylococcus epidermidis.

Staphylococcus epidermidis has become one of the most common causes of septicemia. Meanwhile, S. epidermidis has acquired resistance to many antibiotics. Among these, methicillin-resistant S. epidermidis (MRSE) were frequently isolated. Similar to methicillin resistant Staphylococcus aureus (MRSA), they also exhibited multi-resistance, which presented a danger to human health. Patrinia scabiosaefolia as traditional Chinese medicine had strong antibacterial activity against MRSE. However, the mechanism of P. scabiosaefolia against MRSE is not clear. Here, the morphology of cell wall and cell membrane, production of β-lactamase and PBP2, energy metabolism, antioxidant system were systematically studied. The data showed that P. scabiosaefolia damaged the cell wall and membrane. In addition, β-lactamase, energy metabolism and antioxidant system were involved in mechanisms of P. scabiosaefolia against MRSE. These observations provided new understanding of P. scabiosaefolia against MRSE to control MRSE infections.

Dynamic structural evolution of lignin macromolecules and hemicelluloses during Chinese pine growth

To comprehend the biosynthesis processes of conifers, it is essential to investigate the disparity between the cell wall shape and the interior chemical structures of polymers throughout the development of Chinese pine. In this study, branches of mature Chinese pine were separated according to their growth time (2, 4, 6, 8 and 10 years). The variation of cell wall morphology and lignin distribution was comprehensively monitored by scanning electron microscopy (SEM) and confocal Raman microscopy (CRM), respectively. Moreover, the chemical structures of lignin and alkali-extracted hemicelluloses were extensively characterized by nuclear magnetic resonance (NMR) and gel permeation chromatography (GPC). The thickness of latewood cell walls increased steadily from 1.29 μm to 3.38 μm, and the structure of the cell wall components became more complicated as the growth time increased. Based on the structural analysis, it was found that the content of β-O-4 (39.88–45.44/100 Ar), β-β (3.20–10.02/100 Ar) and β-5 (8.09–15.35/100 Ar) linkages as well as the degree of polymerization of lignin increased with the growth time. The complication propensity increased significantly over 6 years before slowing to a trickle over 8 and 10 years. Furthermore, alkali-extracted hemicelluloses of Chinese pine mainly consist of galactoglucomannans and arabinoglucuronxylan, in which the relative content of galactoglucomannans increased with the growth of the pine, especially from 6 to 10 years.

Triphenylphosphonium-Driven Targeting of Pyrimorph Fragment Derivatives Greatly Improved Its Action on Phytopathogen Mitochondria.

Pyrimorph is a carboxylic acid amide (CAA) fungicide, which shows excellent activity against oomycetes such as pepper phytophthora blight, tomato late blight, and downy mildew of cucumber. It works mainly by inhibiting the biosynthesis of cell wall of oomycetes. However, pyrimorph also shows weak activity of inhibiting mitochondrial complex III, which is the first CAA fungicide found to act on mitochondria. To improve this effect on mitochondria and develop fungicides that may have a novel mechanism of action, in this paper, by disassembling pyrimorph and conjugating the fragments with the mitochondrial-targeted delivery system (triphenylphosphonium), three series of mitochondrial-targeting analogues of pyrimorph were designed and synthesized. The results show that the pyridine-containing 1,1-diaryl is the core module of inhibition mitochondrial function of pyrimorph. Among these conjugates, compound 3b with a short linker showed the highest and broad-spectrum fungicidal activity, strong respiratory inhibition activity, and adenosine 5'-triphosphate synthesis inhibition activity, suggesting its potential as a fungicide candidate. 3b exhibited greatly improved action on mitochondria, such as by destroying the mitochondrial function of pathogens, causing mitochondrial swelling, weakening its influence on cell wall morphology, and so on. More importantly, this study provides a method to strengthen the drugs or pesticides with weak mitochondrial action, which is of special significance for developing mitochondrial bioactive molecules with the novel action mechanism.