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Frontiers in Plant Science 2024Rhizobacteria are capable of inducing defense responses via the expression of pathogenesis-related proteins (PR-proteins) such as chitinases, and many studies have...
Rhizobacteria are capable of inducing defense responses via the expression of pathogenesis-related proteins (PR-proteins) such as chitinases, and many studies have validated the functions of plant chitinases in defense responses. Soybean () is an economically important crop worldwide, but the functional validation of soybean chitinase in defense responses remains limited. In this study, genome-wide characterization of soybean chitinases was conducted, and the defense contribution of three chitinases (GmChi01, GmChi02, or GmChi16) was validated in transgenic lines against the soil-borne pathogen . Compared to the Col-0 and empty vector controls, the transgenic lines with GmChi02 or GmChi16 exhibited fewer chlorosis symptoms and wilting. While GmChi02 and GmChi16 enhanced defense to , GmChi02 was the only one significantly induced by . The observation indicated that plant chitinases may be induced by different rhizobacteria for defense responses. The survey of 37 soybean chitinase gene expressions in response to six rhizobacteria observed diverse inducibility, where only 10 genes were significantly upregulated by at least one rhizobacterium and 9 genes did not respond to any of the rhizobacteria. Motif analysis on soybean promoters further identified not only consensus but also rhizobacterium-specific transcription factor-binding sites for the inducible chitinase genes. Collectively, these results confirmed the involvement of GmChi02 and GmChi16 in defense enhancement and highlighted the diverse inducibility of 37 soybean chitinases encountering and six rhizobacteria.
PubMed: 38405589
DOI: 10.3389/fpls.2024.1341181 -
ACS Omega Feb 2024Microbial degradation of dyes is vital to understanding the fate of dyes in the environment. In this study, a fungal strain A-3 and a bacterial strain L-6, which were...
Microbial degradation of dyes is vital to understanding the fate of dyes in the environment. In this study, a fungal strain A-3 and a bacterial strain L-6, which were identified as and , respectively, had been proven to efficiently degrade crystal violet (CV) dye. The decolorization of CV dye by fungal and bacterial cocultivation was investigated. The results showed that the decolorization rate of cocultures was better than monoculture ( in L-6 (PF), and that of A-3 (AF)). Furthermore, enzymatic analysis further revealed that Lac, MnP, Lip, and NADH-DCIP reductases were involved in the biodegradation of CV dyes. UV-visible spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, and gas chromatography-mass spectrometry (GC-MS) were used to examine the degradation products. GC-MS analysis showed the presence of 4-(dimethylamino) benzophenone, 3-dimethylaminophenol, benzyl alcohol, and benzaldehyde, indicating that CV was degraded into simpler compounds. The phytotoxicity tests revealed that CV degradation products were less toxic than the parent compounds, indicating that the cocultures detoxified CV dyes. As a result, the cocultures are likely to have a wide range of applications in the bioremediation of CV dyes.
PubMed: 38405495
DOI: 10.1021/acsomega.3c06978 -
Antonie Van Leeuwenhoek Feb 2024In the search of new enzymatic activities with a possible industrial application, we focused on those microorganisms and their molecular mechanisms that allow them to...
In the search of new enzymatic activities with a possible industrial application, we focused on those microorganisms and their molecular mechanisms that allow them to succeed in the environment, particularly in the proteolytic activity and its central role in the microorganisms' successful permanence. The use of highly active serine proteases for industrial applications is a modern need, especially for the formulation of detergents, protein processing, and hair removal from animal skins. This report provides the isolation and identification of a highly proteolytic fragment derived from DegQ produced by a Pseudomonas fluorescens environmental strain isolated from a frog carcass. Zymograms demonstrate that a 10 kDa protein mainly generates the total proteolytic activity of this strain, which is enhanced by the detergent SDS. Mass spectroscopy analysis revealed that the protein derived a couple of peptides, the ones showing the highest coverage belonging to DegQ. Interestingly, this small protein fragment contains a PDZ domain but no obvious residues indicating that it is a protease. Protein model analysis shows that this fragment corresponds to the main PDZ domain from DegQ, and its unique sequence and structure render a proteolytic peptide. The results presented here indicate that a novel DegQ fragment is sufficient for obtaining high protease activity highlighting that the analysis of environmental microorganisms can render new strains or enzymes with helpful biotechnological characteristics.
Topics: Animals; PDZ Domains; Pseudomonas; Serine Endopeptidases; Peptides; Serine Proteases
PubMed: 38400879
DOI: 10.1007/s10482-024-01939-z -
Microorganisms Feb 2024This study aimed to explore the phenotype and relationship of drug resistance genes in livestock and poultry farm wastewater and drinking water reservoirs to provide...
This study aimed to explore the phenotype and relationship of drug resistance genes in livestock and poultry farm wastewater and drinking water reservoirs to provide evidence for the transmission mechanisms of drug resistance genes, in order to reveal the spread of drug resistance genes in wastewater from intensive farms in Central China to urban reservoirs that serve as drinking water sources and provide preliminary data for the treatment of wastewater from animal farms to reduce the threat to human beings. DNA extraction and metagenomic sequencing were performed on eight groups of samples collected from four water reservoirs and four related wastewaters from animal farms in Central China. Metagenomic sequencing showed that the top 20 AROs with the highest abundance were _gene, _gene, , , , _gene_, _gene, , _gene, _gene, , , , , _gene, , _gene, , , and . The resistance genes mentioned above belong to the following categories of drug resistance mechanisms: antibiotic target replacement, antibiotic target protection, antibiotic inactivation, and antibiotic efflux. The resistomes that match the top 20 genes are and ; ; ; and . ; ; and ; and ; , , , , , , , , , and ; and ; , , , , , and ; and ; and ; , , and ; , , , , , , and . Unreported drug resistance genes and drug-resistant bacteria in Central China were identified in 2023. In the transmission path of drug resistance genes, the transmission path from aquaculture wastewater to human drinking water sources cannot be ignored. For the sake of human health and ecological balance, the treatment of aquaculture wastewater needs to be further strengthened, and the effective blocking of drug resistance gene transmission needs to be considered.
PubMed: 38399800
DOI: 10.3390/microorganisms12020396 -
International Journal of Molecular... Feb 2024(MI) colonizing metalworking fluids (MWFs) has been associated with chronic hypersensitivity pneumonitis (HP) in machinists. However, it is etiologically unclear why...
Differential Immunogenicity and Lung Disease-Inducing Potential of Genotypes and Impact of Co-Exposure with Pseudomonas: Optimizing a Mouse Model of Chronic Hypersensitivity Pneumonitis.
(MI) colonizing metalworking fluids (MWFs) has been associated with chronic hypersensitivity pneumonitis (HP) in machinists. However, it is etiologically unclear why only certain mycobacteria-contaminated fluids induce this interstitial lung disease. We hypothesized that this may be due to differential immunogenicity and the HP-inducing potential of MI strains/genotypes as well as the confounding effect of co-inhaled endotoxin-producers. To test this hypothesis, we optimized a chronic HP mouse model in terms of MI antigen dose, timepoint of sacrifice, and form of antigen (cell lysates vs. live cells) and compared six different field-isolated MI strains. Overall, MJY10 was identified as the most immunogenic and MJY4 (or MJY13) as the least immunogenic genotype based on lung pathoimmunological changes as well as Th1 cellular response (IFN-γ release). Infection with MI live cells induced a more severe phenotype than MI cell lysate. Co-exposure with caused a greater degree of lung innate immune response and granuloma formation but a diminished adaptive (Th1) immune response (IFN-γ) in the lung and spleen. In summary, this study led to the first demonstration of differential immunogenicity and the disease-inducing potential of field strains of MI and an interfering effect of the co-contaminating Pseudomonas. The improved chronic MI-HP mouse model and the identified polar pair of MI strains will facilitate future diagnostic and therapeutic research on this poorly understood environmental lung disease.
Topics: Mice; Animals; Pseudomonas; Alveolitis, Extrinsic Allergic; Lung; Genotype; Mycobacteriaceae
PubMed: 38396736
DOI: 10.3390/ijms25042058 -
Antibiotics (Basel, Switzerland) Jan 2024Industrial biocides aim to keep water systems microbiologically controlled and to minimize biofouling. However, the resulting dead cells are usually not removed from the...
Industrial biocides aim to keep water systems microbiologically controlled and to minimize biofouling. However, the resulting dead cells are usually not removed from the water streams and can influence the growth of the remaining live cells in planktonic and sessile states. This study aims to understand the effect of dead cells killed by industrial biocides-benzalkonium chloride (BAC) and 2,2-dibromo-3-nitrilopropionamide (DBNPA)-on biofilm formation. Additionally, the effect of different dead/live cell ratios (50.00% and 99.99%) was studied. The inoculum was recirculated in a Parallel Plate Flow Cell (PPFC). The overall results indicate that dead cells greatly affect biofilm properties. Inoculum with DBNPA-dead cells led to more active (higher ATP content and metabolic activity) and thicker biofilm layers in comparison to BAC-dead cells, which seems to be linked to the mechanism of action by which the cells were killed. Furthermore, higher dead cell ratios (99.99%) in the inoculum led to more active (higher culturability, metabolic activity and ATP content) and cohesive/compact and uniformly distributed biofilms in comparison with the 50.00% dead cell ratio. The design of future disinfection strategies must consider the contribution of dead cells to the biofilm build-up, as they might negatively affect water system operations.
PubMed: 38391526
DOI: 10.3390/antibiotics13020140 -
Journal of Inherited Metabolic Disease Feb 2024Phenylketonuria (PKU) is a congenital metabolic disorder that causes the systemic elevation of phenylalanine (Phe), which is neurotoxic and teratogenic. PKU is currently...
Phenylketonuria (PKU) is a congenital metabolic disorder that causes the systemic elevation of phenylalanine (Phe), which is neurotoxic and teratogenic. PKU is currently incurable, and management involves lifelong adherence to an unpalatable protein-restricted diet based on Phe-free amino acid mixtures. Seeking a palatable dietary alternative, we identified a Bacillus subtilis protein (GSP16O) with a well-balanced but low-Phe amino acid profile. We optimized the sequence and expressed a modified Phe-free version (GSP105) in Pseudomonas fluorescens, achieving yields of 20 g/L. The purified GSP105 protein has a neutral taste and smell, is highly soluble, and remains stable up to 80°C. Homozygous enu2 mice, a model of human PKU, were fed with diets containing either GSP105 or normal protein. The GSP105 diet led to normalization of blood Phe levels and brain monoamine neurotransmitter metabolites, and prevented maternal PKU. The GSP105 diet thus provides an alternative and efficacious dietary management strategy for PKU.
PubMed: 38390655
DOI: 10.1002/jimd.12719 -
Analytical Chemistry Mar 2024Gas chromatography combined with ion mobility spectrometry (GC-IMS) is a powerful separation and detection technique for volatile organic compounds (VOC). This...
Gas chromatography combined with ion mobility spectrometry (GC-IMS) is a powerful separation and detection technique for volatile organic compounds (VOC). This combination is characterized by exceptionally low detection limits in the low ppbv range, high 2-dimensional selectivity, and robust operation. These qualities make it an ideal tool for nontarget screening approaches. Fermentation broths contain a substantial number of VOC, either from the medium or produced by microbial metabolism, that are currently not regularly measured for process monitoring. In this study, , , , and were exemplarily used as model organisms and cultivated, and the headspace was analyzed by GC-IMS. Additionally, mixed cultures for every combination of two of the microorganisms were also characterized. Multivariate data analysis of the GC-IMS data revealed that it is possible to differentiate between the microorganisms using PLS-DA with a prediction accuracy of 0.92. The mixed cultures could be separated from the pure cultures with accuracies between 0.87 and 1.00 depending on the organism. GC-IMS data correlate with the optical density and can be used to follow and model growth curves. The root mean squared errors ranged between 10 and 20% of the maximum value, depending on the organism. Peak identification with reference compounds did not reveal specific marker compounds, rather the pattern was found to be responsible for the model performance.
Topics: Ion Mobility Spectrometry; Gas Chromatography-Mass Spectrometry; Volatile Organic Compounds; Fermentation; Multivariate Analysis; Escherichia coli
PubMed: 38386844
DOI: 10.1021/acs.analchem.3c04857 -
Scientific Reports Feb 2024Microbial inoculants are attracting growing interest in agriculture, but their efficacy remains unreliable in relation to their poor survival, partly due to the...
Microbial inoculants are attracting growing interest in agriculture, but their efficacy remains unreliable in relation to their poor survival, partly due to the competition with the soil resident community. We hypothesised that recurrent inoculation could gradually alleviate this competition and improve the survival of the inoculant while increasing its impact on the resident bacterial community. We tested the effectiveness of such strategy with four inoculation sequences of Pseudomonas fluorescens strain B177 in soil microcosms with increasing number and frequency of inoculation, compared to a non-inoculated control. Each sequence was carried out at two inoculation densities (10 and 10 cfu.g soil). The four-inoculation sequence induced a higher abundance of P. fluorescens, 2 weeks after the last inoculation. No impact of inoculation sequences was observed on the resident community diversity and composition. Differential abundance analysis identified only 28 out of 576 dominants OTUs affected by the high-density inoculum, whatever the inoculation sequence. Recurrent inoculations induced a strong accumulation of nitrate, not explained by the abundance of nitrifying or nitrate-reducing microorganisms. In summary, inoculant density rather than inoculation pattern matters for inoculation effect on the resident bacterial communities, while recurrent inoculation allowed to slightly enhance the survival of the inoculant and strongly increased soil nitrate content.
Topics: Agricultural Inoculants; Soil; Nitrates; Agriculture; Pseudomonas fluorescens; Soil Microbiology
PubMed: 38378706
DOI: 10.1038/s41598-024-54069-x -
MicroLife 2024Pyoverdin is a water-soluble metal-chelator synthesized by members of the genus and used for the acquisition of insoluble ferric iron. Although freely diffusible in...
Pyoverdin is a water-soluble metal-chelator synthesized by members of the genus and used for the acquisition of insoluble ferric iron. Although freely diffusible in aqueous environments, preferential dissemination of pyoverdin among adjacent cells, fine-tuning of intracellular siderophore concentrations, and fitness advantages to pyoverdin-producing versus nonproducing cells, indicate control of location and release. Here, using time-lapse fluorescence microscopy to track single cells in growing microcolonies of SBW25, we show accumulation of pyoverdin at cell poles. Accumulation occurs on cessation of cell growth, is achieved by cross-feeding in pyoverdin-nonproducing mutants and is reversible. Moreover, accumulation coincides with localization of a fluorescent periplasmic reporter, suggesting that pyoverdin accumulation at cell poles is part of the general cellular response to starvation. Compatible with this conclusion is absence of non-accumulating phenotypes in a range of pyoverdin mutants. Analysis of the performance of pyoverdin-producing and nonproducing cells under conditions promoting polar accumulation shows an advantage to accumulation on resumption of growth after stress. Examination of pyoverdin polar accumulation in a multispecies community and in a range of laboratory and natural species of , including PAO1 and KT2440, confirms that the phenotype is characteristic of .
PubMed: 38370141
DOI: 10.1093/femsml/uqae001