Expression of <i>rsmY</i> and <i>rsmZ</i> was dramatically increased under stressful circumstances.

<p>Expression levels of genes were detected in WT PAO1, which was cultivated in (A) LB broth medium (B) MH-S cells as well as (C) LB broth medium containing 2<b> </b>µg/ml gentamicin at different time-points. Data are means ± SEM, and representative of three experiments. *, <i>P</i><0.05; **, <i>P</i><0.01, ***, <i>P</i><0.001, one-way ANOVA (Tukey-Kramer post hoc).</p

RsmYZ regulated cooperation was induced by environmental nutrient factors.

<p>(A) Growth of WT PAO1 in M9<b> </b>minimal growth medium containing 1% adenosine (circle), 1% adenosine and 1% BSA as the dual carbon source (black square) and BSA was added after 40 hours cultivation (triangle). Expression of leadership and dependent genes of WT PAO1 were detected in M9<b> </b>minimal growth medium containing (B) 1% adenosine; (C) 0.5% adenosine+0.5% BSA; (D) 0.5% +0.5% BSA and the supernatants were removed every 4 hours; (E) 1% adenosine and then 1% BSA was added after 20 hours cultivation; (F) 1% adenosine and then 1% BSA was added after 40 hours cultivation. Data are means ± SEM, and representative of three experiments. *, <i>P</i><0.05; **, <i>P</i><0.01; ***, <i>P</i><0.001, one-way ANOVA (Tukey-Kramer post hoc).</p

GacA-RsmYZ leadership cascade governs the initial expression of the QS-dependent genes.

<p>Gene expression of leadership and QS regulators in WT PAO1 (A) and QS-deficient strain PAO1-<i>ΔlasR</i> (B) in low density (OD₆₀₀≈0.12), quorum density (OD₆₀₀≈0.28), and high density (OD₆₀₀≈0.9) phases. Data are means ± standard error means (SEM), and representative of three experiments. *, <i>P</i><0.01; **, <i>P</i><0.001, one-way ANOVA (Tukey-Kramer post hoc).</p

High levels of <i>rsmY</i> and <i>rsmZ</i> expression could cause an enhanced biofilm production after 24 h.

<p>Production of biofilm was detected by crystal violet staining (A) and quantified at OD₅₉₅ (B). The data are represented as means ± SEM, six replicates per culture. *, <i>P</i><0.05, one-way ANOVA (Tukey-Kramer post hoc).</p

Cooperation activation was delayed due to the presence of <i>lasR</i> mutant.

<p>(A) The same amounts of wild type (WT) <i>P. aeruginosa</i> PAO1 were cultured in different volumes of LB broth medium. (B) 10<b> </b>µl of overnight cultivated WT PAO1, 5<b> </b>µl WT PAO1+5<b> </b>µl PAO1-<i>ΔlasR</i>, and 10<b> </b>µl PAO1-<i>ΔlasR</i> strains were independently cultured in the same volume of LB broth medium. Activation of cooperation (☆) was determined by significantly increased expression of <i>lasB</i> gene. Data are means ± SEM, and representative of three experiments. *, <i>P</i><0.05, one-way ANOVA (Tukey-Kramer post hoc).</p

Data_Sheet_3_Unraveling the antimicrobial potential of Lactiplantibacillus plantarum strains TE0907 and TE1809 sourced from Bufo gargarizans: advancing the frontier of probiotic-based therapeutics.xlsx

IntroductionIn an era increasingly defined by the challenge of antibiotic resistance, this study offers groundbreaking insights into the antibacterial properties of two distinct Lactiplantibacillus plantarum strains, TE0907 and TE1809, hailing from the unique ecosystem of Bufo gargarizans. It uniquely focuses on elucidating the intricate components and mechanisms that empower these strains with their notable antibacterial capabilities.MethodsThe research employs a multi-omics approach, including agar diffusion tests to assess antibacterial efficacy and adhesion assays with HT-29 cells to understand the preliminary mechanisms. Additionally, gas chromatography-mass spectrometry (GC-MS) is employed to analyze the production of organic acids, notably acetic acid, and whole-genome sequencing is utilized to identify genes linked to the biosynthesis of antibiotics and bacteriocin-coding domains.ResultsThe comparative analysis highlighted the exceptional antibacterial efficacy of strains TE0907 and TE1809, with mean inhibitory zones measured at 14.97 and 15.98 mm, respectively. A pivotal discovery was the significant synthesis of acetic acid in both strains, demonstrated by a robust correlation coefficient (cor ≥ 0.943), linking its abundance to their antimicrobial efficiency. Genomic exploration uncovered a diverse range of elements involved in the biosynthesis of antibiotics similar to tetracycline and vancomycin and potential regions encoding bacteriocins, including Enterolysin and Plantaricin.ConclusionThis research illuminates the remarkable antibacterial efficacy and mechanisms intrinsic to L. plantarum strains TE0907 and TE1809, sourced from B. gargarizans. The findings underscore the strains' extensive biochemical and enzymatic armamentarium, offering valuable insights into their role in antagonizing enteric pathogens. These results lay down a comprehensive analytical foundation for the potential clinical deployment of these strains in safeguarding animal gut health, thereby enriching our understanding of the role of probiotic bacteria in the realm of antimicrobial interventions.</p

Image_1_Unraveling the antimicrobial potential of Lactiplantibacillus plantarum strains TE0907 and TE1809 sourced from Bufo gargarizans: advancing the frontier of probiotic-based therapeutics.jpeg

IntroductionIn an era increasingly defined by the challenge of antibiotic resistance, this study offers groundbreaking insights into the antibacterial properties of two distinct Lactiplantibacillus plantarum strains, TE0907 and TE1809, hailing from the unique ecosystem of Bufo gargarizans. It uniquely focuses on elucidating the intricate components and mechanisms that empower these strains with their notable antibacterial capabilities.MethodsThe research employs a multi-omics approach, including agar diffusion tests to assess antibacterial efficacy and adhesion assays with HT-29 cells to understand the preliminary mechanisms. Additionally, gas chromatography-mass spectrometry (GC-MS) is employed to analyze the production of organic acids, notably acetic acid, and whole-genome sequencing is utilized to identify genes linked to the biosynthesis of antibiotics and bacteriocin-coding domains.ResultsThe comparative analysis highlighted the exceptional antibacterial efficacy of strains TE0907 and TE1809, with mean inhibitory zones measured at 14.97 and 15.98 mm, respectively. A pivotal discovery was the significant synthesis of acetic acid in both strains, demonstrated by a robust correlation coefficient (cor ≥ 0.943), linking its abundance to their antimicrobial efficiency. Genomic exploration uncovered a diverse range of elements involved in the biosynthesis of antibiotics similar to tetracycline and vancomycin and potential regions encoding bacteriocins, including Enterolysin and Plantaricin.ConclusionThis research illuminates the remarkable antibacterial efficacy and mechanisms intrinsic to L. plantarum strains TE0907 and TE1809, sourced from B. gargarizans. The findings underscore the strains' extensive biochemical and enzymatic armamentarium, offering valuable insights into their role in antagonizing enteric pathogens. These results lay down a comprehensive analytical foundation for the potential clinical deployment of these strains in safeguarding animal gut health, thereby enriching our understanding of the role of probiotic bacteria in the realm of antimicrobial interventions.</p

Table <b>5.</b> Results of hierarchical analyses of molecular variance (AMOVA) conducted using different grouping options.

*<p>Populations contain only one sample are exclude in all the analyses Main evolutionary clades and subclades in clade 1 are observed in our phylogenetic trees.</p><p># Jiajin Moutain, Erlang Moutain, Gongga Mountain, Hongya county.</p

Result of the Bayesian coalescent-based estimation of divergence time among different evolutionary clades/subclades and the time to the most recent common ancestor (<i>t</i>_MRCA) of different evolutionary clades/subclades.

<p>Results based on non-partition and partitioned by each codon were shown (two partitions: pos1+pos2, pos3; three partitions: pos1, pos2, pos3). Mean values are shown in bold and 95% HPD are shown in brackets.</p

Data_Sheet_2_Unraveling the antimicrobial potential of Lactiplantibacillus plantarum strains TE0907 and TE1809 sourced from Bufo gargarizans: advancing the frontier of probiotic-based therapeutics.xlsx

IntroductionIn an era increasingly defined by the challenge of antibiotic resistance, this study offers groundbreaking insights into the antibacterial properties of two distinct Lactiplantibacillus plantarum strains, TE0907 and TE1809, hailing from the unique ecosystem of Bufo gargarizans. It uniquely focuses on elucidating the intricate components and mechanisms that empower these strains with their notable antibacterial capabilities.MethodsThe research employs a multi-omics approach, including agar diffusion tests to assess antibacterial efficacy and adhesion assays with HT-29 cells to understand the preliminary mechanisms. Additionally, gas chromatography-mass spectrometry (GC-MS) is employed to analyze the production of organic acids, notably acetic acid, and whole-genome sequencing is utilized to identify genes linked to the biosynthesis of antibiotics and bacteriocin-coding domains.ResultsThe comparative analysis highlighted the exceptional antibacterial efficacy of strains TE0907 and TE1809, with mean inhibitory zones measured at 14.97 and 15.98 mm, respectively. A pivotal discovery was the significant synthesis of acetic acid in both strains, demonstrated by a robust correlation coefficient (cor ≥ 0.943), linking its abundance to their antimicrobial efficiency. Genomic exploration uncovered a diverse range of elements involved in the biosynthesis of antibiotics similar to tetracycline and vancomycin and potential regions encoding bacteriocins, including Enterolysin and Plantaricin.ConclusionThis research illuminates the remarkable antibacterial efficacy and mechanisms intrinsic to L. plantarum strains TE0907 and TE1809, sourced from B. gargarizans. The findings underscore the strains' extensive biochemical and enzymatic armamentarium, offering valuable insights into their role in antagonizing enteric pathogens. These results lay down a comprehensive analytical foundation for the potential clinical deployment of these strains in safeguarding animal gut health, thereby enriching our understanding of the role of probiotic bacteria in the realm of antimicrobial interventions.</p