Journal: Current Research in Microbial Sciences

Article Title: The glutathione pathway is required for biofilm formation in Acinetobacter baumannii

doi: 10.1016/j.crmicr.2026.100562

Figure Lengend Snippet: Growth rates of A. baumannii gsh mutant strains in liquid tryptic soy broth media. Strains were grown in triplicate in 10 mL media in 50 mL conical tubes. Logarithmically growing cultures were diluted to 0.05 OD 600 in TSB and incubated at 37 °C with shaking. Aliquots were taken at specific timepoints and (a) OD 600 and (b) CFUs were determined; wildtype (black solid line, closed square), WTev (black dashed line, open square), gshA − ( red solid line, closed square), gshA-C (red dashed line, open square), gshB − (blue solid line, closed square), gshB-C (blue dashed line, open square). Error bars indicate standard deviation calculated from three independent biological replicates at each timepoint.

Article Snippet: Upregulation of the phenylacetate (PAA) catabolic pathway aligns with reduced virulence observed in ATCC 17978 gshA mutant strains in the Galleria mellonella infection model.

Techniques: Mutagenesis, Incubation, Standard Deviation

Journal: Current Research in Microbial Sciences

Article Title: The glutathione pathway is required for biofilm formation in Acinetobacter baumannii

doi: 10.1016/j.crmicr.2026.100562

Figure Lengend Snippet: Biofilm formation in A. baumannii gsh and gsnoR mutant strains. (a) Biofilm dry weight assay in tryptic soy broth, (b) biofilm optical density assay in tryptic soy broth, (c) biofilm optical density assay in LB. A. baumannii gshA, gshB, gsnoR1 , and gsnoR2 mutant, complemented, and isogenic wildtype strains were examined. The assay was performed with three independent biological replicates, each with two technical replicates. Data are presented as mean ± standard error of the mean. Statistical significance was determined using an unpaired Student’s t -test. P -values are indicated as * = 0.05, ** = 0.01, *** = 0.001, **** = 0.0001.

Article Snippet: Upregulation of the phenylacetate (PAA) catabolic pathway aligns with reduced virulence observed in ATCC 17978 gshA mutant strains in the Galleria mellonella infection model.

Techniques: Mutagenesis

Journal: Current Research in Microbial Sciences

Article Title: The glutathione pathway is required for biofilm formation in Acinetobacter baumannii

doi: 10.1016/j.crmicr.2026.100562

Figure Lengend Snippet: Confocal scanning laser microscopy (CSLM) analysis of biofilms. (a) Top-views (upper) and side-views (lower) CSLM images of representative biofilms formed by A. baumannii strains: WT, WTev, gshA − , gshA-C, gshB − , gshB-C, gsnoR1 − , gsnoR1-C, gsnoR2 − , and gsnoR2-C. Biofilms were grown in tryptic soy broth in 12-well plates, stained with Syto13 nucleic acid stain, and imaged using a Zeiss LSM 880 confocal microscope using a 488/561 nm diode laser. Z-stacks were acquired at 652 × 652 pixels with 0.5 μm intervals using a 40× water-dipping objective. Images were processed in ImageJ using the "Project Stacks" function to generate top and side views. Scale bars = 25 μm. (b) The area of clumping (µm²) of the strains. Aggregate areas were quantified from the CSLM images by selecting three clumped aggregates from three independent fields of view per strain. The gshA − mutant formed significantly larger aggregates compared to the WT and its complemented strain. Data plotted represents mean aggregate area, and error bars indicate standard deviation. Statistical analysis comparing each mutant strain to its respective complemented strain is shown and was performed by one-way ANOVA. ns, not significant; **** P < 0.0001.

Article Snippet: Upregulation of the phenylacetate (PAA) catabolic pathway aligns with reduced virulence observed in ATCC 17978 gshA mutant strains in the Galleria mellonella infection model.

Techniques: Microscopy, Staining, Mutagenesis, Standard Deviation

Journal: Applied and Environmental Microbiology

Article Title: Carbonic anhydrases contribute to mitochondrial function, conidial development, and pathogenicity of Magnaporthe oryzae

doi: 10.1128/aem.02488-25

Figure Lengend Snippet: MoCA2, MoCA4, and MoCA6 subcellular localization, interaction with MoCA1, and function in Magnaporthe oryzae . ( A ) Subcellular localization in hyphae and conidia. Red fluorescence signals of MoCA2-RFP, MoCA4-RFP, and MoCA6-RFP protein of hyphae and conidia were examined by confocal microscopy and colocalized with Mito-Tracker Green on mitochondria. Scale bar = 10 μm. ( B ) Y2H between MoCA2, MoCA4, MoCA6, and MoCA1. MoCA2-AD, MoCA4-AD, MoCA6-AD, and MoCA1-BD were co-transformed into yeast strain Y2H Gold and cultured on SD−Leu−Trp and SD−Leu−Trp−His media. pGADT7-MoCA2/pGBKT7-MoCA1, pGADT7-MoCA4/pGBKT7-MoCA1, and pGADT7-MoCA6/pGBKT7-MoCA1, experimental group; pGADT7-T/pGBKT7-53 , positive control group; pGADT7/pGBKT7-MoCA1, negative control group; pGADT7-T/pGBKT7-lam , blank group. ( C ) BiFC assay demonstrating the interaction between MoCA2, MoCA4, MoCA6, and MoCA1. YFP fluorescence signals were observed in N. benthamiana leaves following co-expression of MoCA2-YC, MoCA4-YC, and MoCA6-YC (C-terminal fragment of YFP) with MoCA1-YN (N-terminal fragment of YFP). JC-1 staining confirmed mitochondrial membrane localization. Negative controls included MoCA2-YC, MoCA4-YC, MoCA6-YC, and YN alone. Scale bar = 5 μm.

Article Snippet: Wild-type and MoCA1 , MoCA2 , MoCA4 , MoCA6 knockout mutant strains were cultured in the PDA medium containing CA inhibitor-acetazolamide (Ace) (MCE, China) ( ) at a concentration of 50 nM in the experimental group and 0 nM in the control group at a temperature of 28°C, and the lesion area was counted on the 7th day.

Techniques: Fluorescence, Confocal Microscopy, Transformation Assay, Cell Culture, Positive Control, Negative Control, Bimolecular Fluorescence Complementation Assay, Expressing, Staining, Membrane

Journal: Applied and Environmental Microbiology

Article Title: Carbonic anhydrases contribute to mitochondrial function, conidial development, and pathogenicity of Magnaporthe oryzae

doi: 10.1128/aem.02488-25

Figure Lengend Snippet: Analysis of synergistic effects of MoCA genes. ( A ) Colony morphology of WT, ∆ MoCA1 , and ∆ MoCA2 , ∆ MoCA4 , and ∆ MoCA6 on media with 0 nM and 50 nM carbonic anhydrase inhibitor. ( B ) Colony area measurement and statistical analysis. ( C ) Statistical analysis of MoCA gene expression in ACE of the WT strain. ( D ) Statistical analysis of MoCA1 gene expression in WT, Δ MoCA2 , Δ MoCA4 , and Δ MoCA6 . ( E ) Statistical analysis of MoCA2 gene expression in WT, Δ MoCA1 , Δ MoCA4 , and Δ MoCA6 . ( F ) Statistical analysis of MoCA4 gene expression in WT, Δ MoCA1 , Δ MoCA2 , and Δ MoCA6 . ( G ) Statistical analysis of MoCA6 gene expression in WT, Δ MoCA1 , Δ MoCA2 , and Δ MoCA4 . Error bars represent mean ± SD from three replicates. ns, P > 0.05; * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Wild-type and MoCA1 , MoCA2 , MoCA4 , MoCA6 knockout mutant strains were cultured in the PDA medium containing CA inhibitor-acetazolamide (Ace) (MCE, China) ( ) at a concentration of 50 nM in the experimental group and 0 nM in the control group at a temperature of 28°C, and the lesion area was counted on the 7th day.

Techniques: Gene Expression

Journal: Applied and Environmental Microbiology

Article Title: Carbonic anhydrases contribute to mitochondrial function, conidial development, and pathogenicity of Magnaporthe oryzae

doi: 10.1128/aem.02488-25

Figure Lengend Snippet: Pathogenicity and invasive hyphal growth mechanisms of MoCA2 , MoCA4 , and MoCA6 . ( A ) Spray inoculation assay on rice leaves. Conidia suspensions of the WT, Δ MoCAs , and Δ MoCAs / CAs were used to infect rice leaves. ( B ) Lesion area analysis. ( C ) Statistical analysis of relative fungal growth. ( D ) Rice leaf sheath infection assay. The conidial suspension of indicated strains was injected into a rice sheath. Representative photographs of infectious hyphae were taken after 24, 48, and 72 h of incubation at 25°C. Scale bar = 10 μm. IH, infectious hyphae. ( E ) The infection rate was calculated according to the number of type 1 to type 4 events. The infection status of more than 100 germinated conidia per leaf sheath was scored at 24, 48, and 72 hpi. Type 1 has no penetration; type 2 only has a penetration peg or a single infectious hypha (IH); type 3 has more than two IH in one rice cell; type 4 has extensive IH into adjacent rice cells. Error bars represent at least three independently repeated standard deviations. Error bars represent at least three independently repeated standard deviations. ns, P > 0.05, * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Wild-type and MoCA1 , MoCA2 , MoCA4 , MoCA6 knockout mutant strains were cultured in the PDA medium containing CA inhibitor-acetazolamide (Ace) (MCE, China) ( ) at a concentration of 50 nM in the experimental group and 0 nM in the control group at a temperature of 28°C, and the lesion area was counted on the 7th day.

Techniques: Infection, Suspension, Injection, Incubation

Journal: Applied and Environmental Microbiology

Article Title: Carbonic anhydrases contribute to mitochondrial function, conidial development, and pathogenicity of Magnaporthe oryzae

doi: 10.1128/aem.02488-25

Figure Lengend Snippet: The role of MoCA2 , MoCA4, and MoCA6 in ATP synthesis. ( A ) Mitochondrial membrane potential test using the JC-1 dye. Confocal microscopy was used for observation, with CCCP as a positive control. Scale bar = 10 µm. (B）Ratio of red to green fluorescence intensity indicating mitochondrial membrane potential. ( C ) Statistical analysis of the ATP content of mutant strains. ( D ) Statistical analysis of ATP synthase-related gene expression in WT and knockout strains of M. oryzae . Error bars represent mean ± SD from three replicates. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Wild-type and MoCA1 , MoCA2 , MoCA4 , MoCA6 knockout mutant strains were cultured in the PDA medium containing CA inhibitor-acetazolamide (Ace) (MCE, China) ( ) at a concentration of 50 nM in the experimental group and 0 nM in the control group at a temperature of 28°C, and the lesion area was counted on the 7th day.

Techniques: Membrane, Confocal Microscopy, Positive Control, Fluorescence, Mutagenesis, Gene Expression, Knock-Out

Journal: Applied and Environmental Microbiology

Article Title: Carbonic anhydrases contribute to mitochondrial function, conidial development, and pathogenicity of Magnaporthe oryzae

doi: 10.1128/aem.02488-25

Figure Lengend Snippet: The role of MoCA2, MoCA4, and MoCA6 in nitrogen metabolism. ( A ) Strains inoculated in MM (abundant-nitrogen medium), MM-N (nitrogen-free medium), MM-N (+Gln) (nitrogen-free medium with glutamic acid), and MM-N (+Glu) (nitrogen-free medium with glutamine). The hyphae were stained with Congo red dye and photographed for images. ( B ) Statistical analysis of colony size after 7 days of cultivation in abundant-nitrogen medium, nitrogen-free medium, nitrogen-free medium with glutamic acid, and nitrogen-free medium with glutamine. ( C ) Statistical analysis of gene expression in nitrogen metabolism of WT, Δ MoCA2 , Δ MoCA4 , and Δ MoCA6 . Error bars represent at least three independently repeated standard deviations. ns, P > 0.05; * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Wild-type and MoCA1 , MoCA2 , MoCA4 , MoCA6 knockout mutant strains were cultured in the PDA medium containing CA inhibitor-acetazolamide (Ace) (MCE, China) ( ) at a concentration of 50 nM in the experimental group and 0 nM in the control group at a temperature of 28°C, and the lesion area was counted on the 7th day.

Techniques: Staining, Gene Expression

Journal: mSphere

Article Title: Genomic and phenotypic analysis of ST25 A. baumannii identifies virulence-associated clades and capsular/outer core locus types

doi: 10.1128/msphere.00717-25

Figure Lengend Snippet: Serum resistance of ST25 A. baumannii , ST2 ACICU, ST1 AYE, and ST52 ATCC 19606. The viable cells (CFU/mL) were determined for each isolate following a 5- to 60-min incubation in 20% activated serum and normalized using values obtained from incubation with heat-inactivated. The data were obtained from three independent experiments in which each isolate was tested in triplicate. *** P -values <0.001.

Article Snippet: To investigate the contribution of EPs to desiccation survival in A. baumannii , the ability of ATCC 19606 A. baumannii and ATCC 19606 EPs mutant strains to survive under desiccating conditions was investigated.

Techniques: Incubation