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Genes Oct 2023The model rhizobacterium F113, a relevant plant growth-promoting bacterium, encodes three different Type VI secretion systems (T6SS) in its genome. In silico analysis...
The model rhizobacterium F113, a relevant plant growth-promoting bacterium, encodes three different Type VI secretion systems (T6SS) in its genome. In silico analysis of its genome revealed the presence of a genetic auxiliary module containing a gene encoding an orphan VgrG protein (VgrG5a) that is not genetically linked to any T6SS structural cluster, but is associated with genes encoding putative T6SS-related proteins: a possible adaptor Tap protein, followed by a putative effector, Tfe8, and its putative cognate immunity protein, Tfi8. The bioinformatic analysis of the VgrG5a auxiliary module has revealed that this cluster is only present in several subgroups of the . complex of species. An analysis of the mutants affecting the and genes has shown that the module is involved in bacterial killing. To test whether Tfe8/Tfi8 constitute an effector-immunity pair, the genes encoding Tfe8 and Tfi8 were cloned and expressed in . , showing that the ectopic expression of affected growth. The growth defect was suppressed by ectopic expression. These results indicate that Tfe8 is a bacterial killing effector, while Tfi8 is its cognate immunity protein. The Tfe8 protein sequence presents homology to the proteins of the MATE family involved in drug extrusion. The Tfe8 effector is a membrane protein with 10 to 12 transmembrane domains that could destabilize the membranes of target cells by the formation of pores, revealing the importance of these effectors for bacterial interaction. Tfe8 represents a novel type of a T6SS effector present in pseudomonads.
Topics: Type VI Secretion Systems; Escherichia coli; Bacterial Proteins; Amino Acid Sequence; Pseudomonas
PubMed: 38002922
DOI: 10.3390/genes14111979 -
Nucleic Acids Research Jan 2024The ubiquitous bacterial second messenger cyclic diguanylate (c-di-GMP) coordinates diverse cellular processes through its downstream receptors. However, whether...
The ubiquitous bacterial second messenger cyclic diguanylate (c-di-GMP) coordinates diverse cellular processes through its downstream receptors. However, whether c-di-GMP participates in regulating nitrate assimilation is unclear. Here, we found that NasT, an antiterminator involved in nitrate assimilation in Pseudomonas putida, specifically bound c-di-GMP. NasT was essential for expressing the nirBD operon encoding nitrite reductase during nitrate assimilation. High-level c-di-GMP inhibited the binding of NasT to the leading RNA of nirBD operon (NalA), thus attenuating the antitermination function of NasT, resulting in decreased nirBD expression and nitrite reductase activity, which in turn led to increased nitrite accumulation in cells and its export. Molecular docking and point mutation assays revealed five residues in NasT (R70, Q72, D123, K127 and R140) involved in c-di-GMP-binding, of which R140 was essential for both c-di-GMP-binding and NalA-binding. Three diguanylate cyclases (c-di-GMP synthetases) were found to interact with NasT and inhibited nirBD expression, including WspR, PP_2557, and PP_4405. Besides, the c-di-GMP-binding ability of NasT was conserved in the other three representative Pseudomonas species, including P. aeruginosa, P. fluorescens and P. syringae. Our findings provide new insights into nitrate assimilation regulation by revealing the mechanism by which c-di-GMP inhibits nitrate assimilation via NasT.
Topics: Bacterial Proteins; Cyclic GMP; Gene Expression Regulation, Bacterial; Molecular Docking Simulation; Nitrates; Nitrite Reductases; Pseudomonas aeruginosa; Pseudomonas putida
PubMed: 38000372
DOI: 10.1093/nar/gkad1117 -
ACS ES&T Water Nov 2023This study assesses the efficacy of ultraviolet light-emitting diodes (UV LEDs) for deactivating (pure culture) and (pure culture and biofilms) on relevant drinking...
This study assesses the efficacy of ultraviolet light-emitting diodes (UV LEDs) for deactivating (pure culture) and (pure culture and biofilms) on relevant drinking water distribution system surfaces (cast iron and stainless steel). UV LED treatment at 280 nm demonstrated superior performance compared to that at 365 nm, achieving a 4.8 log reduction value (LRV) for pure cultures and, for biofilms, 4.02 LRV for stainless steel and 2.96 LRV for cast iron at 280 nm. Conversely, the results were less effective at 365 nm, with suspected photolytic reactions on cast iron. Quantification of yielded varying results: 4 LRV using standard plate counts, 1.8 LRV with Legiolert, and 1 LRV with quantitative polymerase chain reaction at 280 nm, while the results were less than 1.5 LRV at 365 nm. This study provides insights into managing opportunistic pathogens and biofilms, emphasizing the need for improved quantification tools to better assess treatment efficacy.
PubMed: 37970541
DOI: 10.1021/acsestwater.3c00428 -
ACS Omega Nov 2023Agronomic management of a crop, including the application of fertilizers and biological inoculants, affects the phenol and flavonoid contents of plants producing these...
Agronomic management of a crop, including the application of fertilizers and biological inoculants, affects the phenol and flavonoid contents of plants producing these metabolites. Kunth, a woody bamboo widely distributed in the Americas, produces several biologically active phenolic compounds. The aim of this study was to evaluate the effect of chemical and organic fertilizers together with the application of biological inoculants on the composition of phenolic compounds in plants at the nursery stage. In 8-month-old plants, differences were observed in plant biomass (20.27 ± 7.68 g) and in the content of total phenols and flavonoids (21.89 ± 9.64 mg gallic acid equivalents/plant and 2.13 ± 0.98 mg quercetin equivalents/plant, respectively) when using the chemical fertilizer diammonium phosphate (DAP). No significant differences were found owing to the effect of the inoculants, although the plants with the application of sp. on plants fertilized with DAP presented higher values of the metabolites (24.12 ± 6.72 mg gallic acid equivalents/plant and 2.39 ± 0.77 mg quercetin equivalents/plant). The chromatographic profile of phenolic metabolites is dominated by one glycosylated flavonoid, the concentration of which was favored by the application of the inoculants , , and sp. In the case study, the combined use of DAP and bacterial inoculants is recommended for the production of plant material with a high content of promising biologically active flavonoids or phenolics.
PubMed: 37970062
DOI: 10.1021/acsomega.3c04579 -
Bactericidal Efficacy and Mechanisms of Non-Electrolytic Slightly Acidic Hypochlorous Water on and .Foods (Basel, Switzerland) Oct 2023Chilled pork is frequently contaminated with and . In this study, the bactericidal efficacy and mechanisms of non-electrolytic slightly acidic hypochlorous water...
Chilled pork is frequently contaminated with and . In this study, the bactericidal efficacy and mechanisms of non-electrolytic slightly acidic hypochlorous water (NE-SAHW) against two strains of these two species were evaluated. The results showed that the antibacterial efficacy of NE-SAHW was positively correlated with the concentration level of NE-SAHW and negatively correlated with the initial populations of the strains. The strains of small populations were completely inhibited when provided with each level of NE-SAHW. The killed cells of were 0.94, 1.39, 4.02, and 5.60 log10 CFU/mL, respectively, and of they were 1.21, 1.52, 4.14, and 5.74 log10 CFU/mL, respectively, when the initial populations of the strains were at high levels (about 7 log10 CFU/mL). Both strains were completely killed within 12 s with the available chlorine concentration (ACC) of 50 mg/L of NE-SAHW. Morphological changes in both cells were observed by using a Scanning Electron Microscope (SEM) and it was discovered that the cell membranes were damaged, which led to the leakage of the intracellular substances, including K+, nucleic acid, and protein. In terms of the Fourier Transform Infrared Spectroscopy (FTIR) results, NE-SAHW destroyed the structures of membrane proteins and cell structure proteins, and influenced the composition of polysaccharides. The bacteria were definitely dead after treatment by NE-SAHW compared to the control according to the results of flow cytometry. These results demonstrated the potential bactericidal property of NE-SAHW when applied to the meat and other food sterilization industries.
PubMed: 37959099
DOI: 10.3390/foods12213980 -
Journal of Dairy Science Apr 2024Biofilm formation is usually affected by many environmental factors, including divalent cations. The purpose of the current work was to analyze how calcium (Ca) affects...
Biofilm formation is usually affected by many environmental factors, including divalent cations. The purpose of the current work was to analyze how calcium (Ca) affects the biofilm formation of dairy Pseudomonas fluorescens isolates by investigating their growth, swarming motility, biofilm-forming capacity, extracellular polymeric substance production, and biofilm structures. Moreover, the regulation mechanism of Ca involved in its biofilm formation was explored through RNA-sequencing analysis. This work revealed that supplementation of 5, 10, 15, and 20 mM Ca significantly reduced the swarming motility of P. fluorescens strains (P.F, P.F, and P.F), but the biofilm-forming ability and polysaccharide production were increased after the supplementation of 5 and 10 mM Ca. By the supplementation of Ca, complex structures with more cell clusters glued together in P. fluorescens P.F biofilms were confirmed by scanning electron microscopy, and increased biomass and coverage of P. fluorescens P.F biofilms were observed by confocal laser scanning microscopy. In addition, RNA-sequencing results showed that P. fluorescens P.F showed a transcriptional response to the supplementation of 10 mM Ca, and a total of 137 genes were significantly expressed. The differential genes were represented in 4 upregulated Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways (nonribosomal peptide structures, quorum sensing, biosynthesis of siderophore group nonribosomal peptides, and phenylalanine metabolism), and 4 downregulated KEGG pathways (flagellar assembly, amino sugar and nucleotide sugar metabolism, nitrotoluene degradation, and cationic antimicrobial peptide resistance). The results indicate that Ca might serve as an enhancer to substantially trigger the biofilm formation of dairy P. fluorescens isolates in the dairy industry.
Topics: Animals; Calcium; Pseudomonas fluorescens; Extracellular Polymeric Substance Matrix; Biofilms; RNA
PubMed: 37949404
DOI: 10.3168/jds.2023-23860 -
Communications Biology Nov 2023In-situ utilization of lunar soil resources will effectively improve the self-sufficiency of bioregenerative life support systems for future lunar bases. Therefore, we...
In-situ utilization of lunar soil resources will effectively improve the self-sufficiency of bioregenerative life support systems for future lunar bases. Therefore, we have explored the microbiological method to transform lunar soil into a substrate for plant cultivation. In this study, five species of phosphorus-solubilizing bacteria are used as test strains, and a 21-day bio-improving experiment with another 24-day Nicotiana benthamiana cultivation experiment are carried out on lunar regolith simulant. We have observed that the phosphorus-solublizing bacteria Bacillus mucilaginosus, Bacillus megaterium, and Pseudomonas fluorescens can tolerate the lunar regolith simulant conditions and dissociate the insoluble phosphorus from the regolith simulant. The phosphorus-solubilizing bacteria treatment improves the available phosphorus content of the regolith simulant, promoting the growth of Nicotiana benthamiana. Here we demonstrate that the phosphorus-solubilizing bacteria can effectively improve the fertility of lunar regolith simulant, making it a good cultivation substrate for higher plants. The results can lay a technical foundation for plant cultivation based on lunar regolith resources in future lunar bases.
Topics: Phosphorus; Nicotiana; Soil; Moon; Bacteria
PubMed: 37945659
DOI: 10.1038/s42003-023-05391-z -
Nucleic Acids Research Dec 2023The widespread Pseudomonas genus comprises a collection of related species with remarkable abilities to degrade plastics and polluted wastes and to produce a broad...
The widespread Pseudomonas genus comprises a collection of related species with remarkable abilities to degrade plastics and polluted wastes and to produce a broad set of valuable compounds, ranging from bulk chemicals to pharmaceuticals. Pseudomonas possess characteristics of tolerance and stress resistance making them valuable hosts for industrial and environmental biotechnology. However, efficient and high-throughput genetic engineering tools have limited metabolic engineering efforts and applications. To improve their genome editing capabilities, we first employed a computational biology workflow to generate a genus-specific library of potential single-stranded DNA-annealing proteins (SSAPs). Assessment of the library was performed in different Pseudomonas using a high-throughput pooled recombinase screen followed by Oxford Nanopore NGS analysis. Among different active variants with variable levels of allelic replacement frequency (ARF), efficient SSAPs were found and characterized for mediating recombineering in the four tested species. New variants yielded higher ARFs than existing ones in Pseudomonas putida and Pseudomonas aeruginosa, and expanded the field of recombineering in Pseudomonas taiwanensisand Pseudomonas fluorescens. These findings will enhance the mutagenesis capabilities of these members of the Pseudomonas genus, increasing the possibilities for biotransformation and enhancing their potential for synthetic biology applications. .
Topics: DNA, Single-Stranded; Gene Editing; Metabolic Engineering; Pseudomonas; Pseudomonas putida
PubMed: 37941137
DOI: 10.1093/nar/gkad1024 -
Biofilm Dec 2023Biofilms are complex microbial communities embedded in extracellular matrix. Pathogens within the biofilm become more resistant to the antibiotics than planktonic...
Biofilms are complex microbial communities embedded in extracellular matrix. Pathogens within the biofilm become more resistant to the antibiotics than planktonic counterparts. Novel strategies are required to encounter biofilms. Exopolysaccharides are one of the major components of biofilm matrix and play a vital role in biofilm architecture. In previous studies, a glycosyl hydrolase, PslG, from was found to be able to inhibit biofilm formation by disintegrating exopolysaccharide in biofilms. Here, we investigate the potential spectrum of PslG homologous protein with anti-biofilm activity. One glycosyl hydrolase from , PslG, exhibits anti-biofilm activities and the key catalytic residues of PslG are conserved with those of PslG. PslG at concentrations as low as 50 nM efficiently inhibits the biofilm formation of and disassemble its preformed biofilm. Furthermore, PslG exhibits anti-biofilm activity on a series of , including and pv. . PslG stays active under various temperatures. Our findings suggest that glycosyl hydrolase PslG has potential to be a broad spectrum inhibitor on biofilm formation of a wide range of .
PubMed: 37928620
DOI: 10.1016/j.bioflm.2023.100155 -
Bioinformation 2023Increased amounts of toxicants may cause sever health issues in humans as well as in aquatic life. Scientists are developing new technologies to combat these problems....
Increased amounts of toxicants may cause sever health issues in humans as well as in aquatic life. Scientists are developing new technologies to combat these problems. Biological methods of detoxification are always beneficial for the environment. Pseudomonas fluorescens is known for its detoxification capacity. In this study stains were isolated from different locations of the Ha'il region, Saudia Arabia. The microbial strain AM-1 displayed resistance to heavy metals (Cr6+, Ni2+, Cd2+, Pb2+) and pesticides (BHC, 2,4-D, Mancozeb) at pollutant levels typical of highly contaminated areas. Additionally, AM-1 exhibited substantial detoxification potential, reducing toxicity by 40.67% for heavy metals and 47.4% for pesticides at 3x concentrations. These findings suggest that the AM-1 strain supports environmental remediation and pollution mitigation. Atomic absorption spectrometry (AAS) results exhibited bioremediation efficiency for metals Cr, Ni, and Pb using immobilized cells of AM-1 isolate, estimated to be 60.57%, 68.4%, and 53.93% respectively. These findings show that AM-1 strain has a potential role in bioremediation of water pollutants and may have future implications in wastewater treatment.
PubMed: 37928494
DOI: 10.6026/97320630019901