Search

Background: Engineering the seed microenvironment with embedded bacteriophages and Plant Growth Promoting Rhizobacteria (PGPR) shows promise for enhancing germination, mitigating biotic and abiotic stressors, and improving resilience under challenging environmental conditions. This study aimed to enhance potato seed germination and control bacterial wilt caused by Ralstonia solanacearum and salinity by using novel technology to encapsulate, preserve, and deliver phage therapy and rhizobacteria., Results: Silk fibroin and trehalose biomaterial combined with the phage P-PSG11 and Pseudomonas lalkuanensis were applied to potato seeds. A pot experiment was conducted to investigate pathogen suppression, salt tolerance, and plant growth enhancement. The combination of silk and trehalose effectively preserved both phage and bacteria for ≥ 8 weeks, maintaining both phage titers and bacterial colony counts. Seeds coated with the P-PSG11 and P. lalkuanensis mixture exhibited the highest germination rate at 93.5%, followed by P. lalkuanensis at 86.3%. In vivo evaluations showed significant increases in root length (72.7%, 61.0%, and 22.5%), plant height (71.5%, 65.1%, and 8.2%), and dry matter (129.1%, 125.7%, and 13.1%) for the P-PSG11 and P. lalkuanensis mixture, P. lalkuanensis, and P-PSG11, respectively. The incidence of wilt was significantly reduced by 88.2% and 81.2%, and salinity was mitigated by 83.3% and 79.2% for the P-PSG11 and P. lalkuanensis mixture and P. lalkuanensis treatment, respectively, compared to the control (p < 0.001). The viability of preserved P-PSG11 and P. lalkuanensis was confirmed after one year using phage titers and bacterial colonies., Conclusion: This innovative approach enhanced plant growth, promoted seed germination, controlled wilt disease, and mitigated soil salinity., Competing Interests: Declarations. Ethics approval and consent to participate: Not applicable. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Background: Pseudomonas spp. have drawn considerable attention due to their rhizospheric abundance and exceptional plant growth-promoting attributes. However, more research is needed on the optimal application methods of Pseudomonas mosselii for rice growth, nutrient accumulation, and yield improvement. This research explored the application of the endophytic bacterium P. mosselii PR5 on rice cultivar BRRI dhan29 with four treatments: control, seedling priming, root drenching, and bacterial cell-free culture (CFC) foliar application., Results: PR5 led to better rice growth, improved nutrient acquisition, and higher yields compared to the control, regardless of the application method used. The highest results in fresh weight of root (146.93 g/pot), shoot (758.98 g/pot), and flag leaf (7.88 g/pot), dry weight of root (42.16 g/pot), shoot (97.32 g/pot), and flag leaf (2.69 g/pot), and grains/panicle (224.67), were obtained from seedling priming treatment, whereas root drenching resulted in maximum plant height (105.67 cm), root length (49.0 cm), tillers/pot (23.7), and panicles/pot (17.67). In all three application methods, rice grain yield per pot was higher in PR5 inoculated treatments, compared to the control. The amount of P, Mg and Zn in the shoot and N, P, Ca, Mg and Si content in the flag leaf was significantly increased along with effective suppression of naturally occurring blast disease in bacterial CFC foliar application, validated by multivariate analysis., Conclusion: Our results indicated that rice seedlings priming with PR5 improved rice growth, yield and nutrient uptake, whereas CFC foliar application significantly increased the concentration of most nutrients in the rice plant and suppressed the naturally occurring rice blast disease. This research highlights the significant potential of P. mosselii PR5 in enhancing rice growth, yield, and nutrient uptake, particularly through seedling priming and CFC foliar application methods., (© 2024. The Author(s).)

Drought stress imposes a serious challenge to cultivate wheat, restricting its growth. Drought reduces the capability of plant to uptake essential nutrients. This causes stunted growth, development and yield. Traditional ways to increase wheat growth under drought stress have shortcomings. Using plant-growth-promoting rhizobacteria (PGPR) has proved feasible and eco-friendly way to enhance wheat growth even under the drought stress. Combining PGPR in consortiums further boosts up their effects. In this study, we have checked the efficacy of drought-tolerant Bacillus halotolerans, Pseudomonas sihuiensis and Bacillus atrophaeus in combination. These strains were allowed to grow on PEG 6000 with concentrations (-0.15, -0.49, -0.73 and - 1.2) Mega Pascal (MPa) alone and in combination. Furthermore, Fourier transmission infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) were used. Their biochemical traits such as solubilization of K, P and Zn and the synthesis of siderophore, indole acetic acid (IAA), protease, amylase, hydrogen cyanide (HCN) and 1-aminocyclopropane-1-carboxylate (ACC) deaminase were done. In addition to this, we investigated the optimum folic acid concentration i.e 150 ppm for wheat against drought stress. We conducted a pot experiment to check the growth-enhancing and drought-mitigating effects of consortium and folic acid alone and in combination. As a result, we found a significantly increased wheat biomass, relative water content (RWC), chlorophyll content, antioxidants including glutathione reductase and total soluble sugars and protein content under all treatments. However, the combined treatment of bacterial consortium and folic acid showed maximum potential to boost wheat growth and survival even under drought. We also investigated the minerals uptake by wheat after the treatments and found maximum nutrient uptake under the co-effect of folic acid and bacterial consortium We believe this is the first study that has investigated the optimal dose of folic acid for wheat. Our research is also novel in that we seek to investigate the effects of folic acid along with a bacterial consortium comprising Bacillus halotolerans, Pseudomonas sihuiensis and Bacillus atrophaeus on wheat grown under the drought stress., (© 2024. The Author(s).)

Bacilli and pseudomonads are among the most well-studied microorganisms commonly found in soil and frequently co-isolated. Isolates from these two genera are frequently used as plant beneficial microorganisms; therefore, their interaction in the plant rhizosphere is relevant for agricultural applications. Despite this, no systematic approach has been employed to assess the coexistence of members from these genera. Here, we screened 720 fluorescent soil isolates for their effects on Bacillus subtilis pellicle formation in two types of media and found a predictor for interaction outcome in Pseudomonas taxonomy. Interactions were context-dependent, and both medium composition and culture conditions strongly influenced interactions. Negative interactions were associated with Pseudomonas capeferrum , Pseudomonas entomophila , and Pseudomonas protegens , and 2,4-diacetylphloroglucinol was confirmed as a strong (but not exclusive) inhibitor of B. subtilis . Non-inhibiting strains were closely related to Pseudomonas trivialis and Pseudomonas lini . Using such a non-inhibiting isolate, Pseudomonas P9&#95;31, which increased B. subtilis pellicle formation demonstrated that the two species were spatially segregated in cocultures. Our study is the first one to propose an overall negative outcome from pairwise interactions between B. subtilis and fluorescent pseudomonads; hence, cocultures comprising members from these groups are likely to require additional microorganisms for coexistence., Importance: There is a strong interest in the microbial ecology field to predict interaction among microorganisms, whether two microbial isolates will promote each other's growth or compete for resources. Numerous studies have been performed based on surveying the available literature or testing phylogenetically diverse sets of species in synthetic communities. Here, a high throughput screening has been performed using 720 Pseudomonas isolates, and their impact on the biofilm formation of Bacillus subtilis was tested. The aim was to determine whether a majority of Pseudomonas will promote or inhibit the biofilms of B. subtilis in the co-cultures. This study reports that Pseudomonas taxonomy is a good predictor of interaction outcome, and only a minority of Pseudomonas isolates promote Bacillus biofilm establishment., Competing Interests: The authors declare no conflict of interest.

Fusarium wilt is an important disease on chili plants caused by the fungus Fusarium oxysporum. This study aims to determine the effectiveness of secondary metabolites from 2 Pseudomonad isolates in controlling Fusarium wilt in chili plants and their effect on chili plant growth. This research was conducted in two stages, namely in vitro, which was carried out in the plant health laboratory and greenhouse, Faculty of Agriculture, Universitas Pembangunan Nasional Veteran Jawa Timur, and in vivo which was carried out in chili plantations known to be endemic to Fusarium wilt in Menganti, Gresik. The research design used in this study was Completely Randomized Design for in vitro testing and Randomized Group Design for in vivo testing by testing 9 treatments repeated 3 times. The treatment tested was control, secondary metabolite application of Pseudomonas isolates PY-122 and PY-142 with concentrations of 20% 30%, 40% and 50%. The variables observed were inhibition of secondary metabolites on the growth of Fusarium fungus, disease incubation period, disease severity index, and plant growth. Sequencing results with 16rRNA gene markers for both Pseudomonas isolates showed similarities to Pseudomonas sp. The results showed that the treatment of secondary metabolite PY-142 with a concentration of 50% showed the highest consistent inhibition. In the greenhouse and field tests, the two isolates slowed down the disease incubation period and development compared to controls. In agronomical observations, the average plant height, number of leaves, root length, and appearance of the first flower on the treated plants were higher and more numerous than the control plants.

Fusarium pseudograminearum , a soil-borne fungus, is the cause of the devastating wheat disease known as wheat crown rot (WCR). The persistence of this pathogen in the soil and crop residues contributes to the increased occurrence and severity of WCR. Therefore, developing effective strategies to prevent and manage WCR is of great importance. In this study, we isolated a bacterial strain, designated as SR9, from the stem of wheat, that exhibited potent antagonistic effects against F. pseudograminearum , as well as the biocontrol efficacy of SR9 on WCR was quantified at 83.99% ± 0.11%. We identified SR9 as Pseudomonas khavaziana and demonstrated its potential as a plant probiotic. SR9 displayed broad-spectrum antagonism against other fungal pathogens, including Neurospora dictyophora , Botrytis californica , and Botryosphaeria dothidea . Whole-genome sequencing analysis revealed that SR9 harbored genes encoding various cell wall-degrading enzymes, cellulases, and lipases, along with antifungal metabolites, which are responsible for its antagonistic activity. Gene knockout and quantitative PCR analyses reveal that phenazine is the essential factor for antagonism. SR9 possessed genes related to auxin synthesis, flagellar biosynthesis, biofilm adhesion, and the chemotaxis system, which play pivotal roles in plant colonization and growth promotion; we also evaluated the effects of SR9 on plant growth in wheat and Arabidopsis . Our findings strongly suggest that SR9 holds great promise as a biocontrol agent for WCR in sustainable agriculture.IMPORTANCEThe escalating prevalence of wheat crown rot, primarily attributed to Fusarium pseudograminearum , necessitates the development of cost-effective and eco-friendly biocontrol strategies. While plant endophytes are recognized for their biocontrol potential, reports on effective strains targeting wheat crown rot are sparse. This study introduces the Pseudomonas khavaziana SR9 strain as an efficacious antagonist to the wheat crown rot pathogen Fusarium pseudograminearum . Demonstrating a significant reduction in wheat crown rot incidence and notable plant growth promotion, SR9 emerges as a key contributor to plant health and agricultural sustainability. Our study outlines a biological approach to tackle wheat crown rot, establishing a groundwork for innovative sustainable agricultural practices., Competing Interests: The authors declare no conflict of interest.

The primary approach to managing biofouling in industrial water systems involves the large-scale use of biocides. It is well-established that biofilms are 'cell factories' that release planktonic cells even when challenged with antimicrobials. The effect of isothiazolinone on the metabolic activity and biomass of mixed Pseudomonas biofilms was monitored in real-time using the CEMS-BioSpec system. The exposure of biofilms to the minimum inhibitory concentration (1.25 mg L -1 ) of biocide did not impact planktonic cell production (log 7.5 CFU mL -1 ), while whole-biofilm metabolic activity and biomass accumulation increased. Only the maximum biocide concentration (80 mg L -1 ) resulted in a change in planktonic cell yields and temporal inhibition of biofilm activity and biomass, a factor that needs due consideration in view of dilution in industrial settings. Interfacing the real-time measurement of metabolic activity and biomass with dosing systems is especially relevant to optimizing the use of biocides in industrial water systems., (© 2024 The Author(s). Environmental Microbiology Reports published by John Wiley & Sons Ltd.)

Bio-transformations refer to the chemical modifications made by an organism on a chemical compound that often involves the interaction of plants with microbes to alter the chemical composition of soil or plant. Integrating bio-transformations and entomopathogenic fungi into litchi cultivation can enhance symbiotic relationships, microbial enzymatic activity in rhizosphere, disease suppression and promote overall plant health. The integration of biological formulations and entomopathogenic fungi can significantly influence growth, nutrient dynamics, physiology, and rhizosphere microbiome of air-layered litchi (Litchi chinensis Sonn.) saplings. Biological modifications included, K-mobilizers, AM fungi, Pseudomonas florescence and Azotobacter chroococcum along with Metarhizium, entomopathogenic fungi have been used. The treatments included, T 1 -Litchi orchard soil + sand (1:1); T 2 -Sand + AM fungi + Azotobacter chroococcum (1:2:1); T 3 -Sand + Pseudomonas florecence + K-mobilizer (1:1:1); T 4 - AM fungi + K-mobilizers (1:1); T 5 , P. Florecence + A. chroococcum + K-mobilizer (1:1:1); T 6 -Sand + P. florecence (1:2) and T 7 -Uninoculated control for field performance. Treatments T 4 -T 6 were further uniformly amended with drenching of Metarrhizium in rhizosphere. T 2 application significantly increased resident microbe survival, total chlorophyll content and root soil ratio in seedlings. A. chroococcum, Pseudomonas, K-mobilizers and AM fungi increased in microbial biomass of 2.59, 3.39, 2.42 and 2.77 times, respectively. Acidic phosphatases, dehydrogenases and alkaline phosphatases were increased in rhizosphere. Leaf nutrients reflected through DOP were considerably altered by T 2 treatment. Based on Eigen value, PCA-induced changes at biological modifications showed maximum total variance. The study inferred that the bio-transformations through microbial inoculants and entomopathogenic fungi could be an encouraging strategy to enhance the growth of plants, health and productivity. Such practices align well with the goals of sustainable agriculture through biological means by reducing dependency on chemical inputs. By delving into these aspects, the research gaps including microbial processes, competitive and symbiotic relationships, resistance in microbes and how complex interactions among bio-transformations, entomopathogenic fungi and microbes can significantly impact the health and productivity of litchi. Understanding and harnessing these interactions can lead to more effective and sustainable farming practices., (© 2024. The Author(s).)

The local ecosystems, fishery and human health are all threatened by water blooms, so effectively controlling water blooms has become an urgent and challenging issue. Biological control of water blooms is given priority due to its low cost, high efficiency and environmental friendliness. In this study, Pseudomonas ZY-1 and Bacillus FY-1, two highly-effective algicidal bacteria strains which are able to indirectly lyse algae by separating and screening from the vigorous water body in the paddy alga of Northeast China were obtained. The two bacterial strains have stronger ability to lyse alga in the bacterial liquid concentration of 10 6 CFU/ml, and the alga-lysing rate on 7 d reached 84.03% and 83.11% respectively. The active substance secreted by ZY-1 is not sensitive to the changes of temperature and pH value, while as FY-1 cell-free filtrate is not stable in high temperature above 50 ℃ and pH of 5, it requires the sun light to have the algaecidal effect. The cell-free filtrates of strains ZY-1 and FY-1 had the best lysis effect on Microcystis aeruginosa cells, and the chlorophyll a content of algae decreased to 0.13 ± 0.02 mg/L and 0.14 ± 0.03 mg/L respectively and the Fv/Fm of Microcystis aeruginosa decreased by 97.22% after 7 days. The algaecidal process of ZY-1 and FY-1 may be that the cell-free filtrate inhibits the photosynthesis of Microcystis aeruginosa, and meanwhile it avoids the regeneration and repair of photosynthesis of algal cells by affecting the gene expression and damaging the repair system of algal cells, so the membrane lipid peroxidation is exacerbated and then the membrane of algal cells is broken, the algal cells can't do normal life activities, and finally the algal cell would be killed. The rice seedlings in the algal liquid treatment group are short and show root dysplasia, few roots and brown roots. After treated with cell-free filtrate of ZY-1 and FY-1, the oxidative damage of the rice is obviously reduced, and the harm from Microcystis aeruginosa is reduced, which has the repair effect to the roots of rice seedlings and its aboveground growth. The cell-free filtrate of FY-1 works better than ZY-1. The bacteria strains of ZY-1 and FY-1 have the indirect algaecide trait, which makes them the potential environmentally-friendly algaecidal bacteria and they show broad application in the agricultural production and the control of water blooms., (© 2024. The Author(s).)

In the last few years, there has been an increasing interest in solutions for sustainable agriculture to reduce negative impacts on the environment resulting from modern agricultural practices. The use of environmentally beneficial bacteria, like Pseudomonas, which can increase plant productivity by reducing growth time, is a promising opportunity for sustainable agriculture. Pseudomonas is a gram-negative bacterium genus, commonly present in soils, plants, and irrigation water. Pseudomonas has a wide range of metabolic routes that could benefit agriculture, such as nutrient uptake, pathogen suppression, heavy metal solubilization, drought tolerance, and high salt concentration tolerance. Pseudomonas may even be proposed as a potential tool for future agriculture on other planets, where the use of microorganisms would be essential for crop development in hostile and inhospitable environments. Hence, the present review discusses the potential use of Pseudomonas in sustainable agriculture on planet Earth and potentially on Mars, highlighting its role in plant growth enhancement and plant protection from pathogenic microorganisms.

The ability of bacteria to colonize diverse environmental niches is often linked to their competence in biofilm formation. It depends on the individual characteristics of a strain, the nature of the colonized surface (abiotic or biotic), or the availability of certain nutrients. Pseudomonas donghuensis P482 efficiently colonizes the rhizosphere of various plant hosts, but a connection between plant tissue colonization and the biofilm formation ability of this strain has not yet been established. We demonstrate here that the potential of P482 to form biofilms on abiotic surfaces and the structural characteristics of the biofilm are influenced by the carbon source available to the bacterium, with glycerol promoting the process. Also, the type of substratum, polystyrene or glass, impacts the ability of P482 to attach to the surface. Moreover, P482 mutants in genes associated with motility or chemotaxis, the synthesis of polysaccharides, and encoding proteases or regulatory factors, which affect biofilm formation on glass, were fully capable of colonizing the root tissue of both tomato and maize hosts. Investigating the role of cellular factors in biofilm formation using these plant-associated bacteria shows that the ability of bacteria to form biofilm on abiotic surfaces does not necessarily mirror its ability to colonize plant tissues. Our research provides a broader perspective on the adaptation of these bacteria to various environments.

Microbiomes often benefit plants, conferring resistance to pathogens, improving stress tolerance, or promoting plant growth. As potential plant mutualists, however, microbiomes are not a single organism but a community of species with complex interactions among microbial taxa and between microbes and their shared host. The nature of ecological interactions among microbes in the microbiome can have important consequences for the net effects of microbiomes on hosts. Here, we compared the effects of individual microbial strains and 10-strain synthetic communities on microbial productivity and host growth using the common duckweed Lemna minor and a synthetic, simplified version of its native microbiome. Except for Pseudomonas protegens , which was a mutualist when tested alone, all of the single strains we tested were commensals on hosts, benefiting from plant presence but not increasing host growth relative to uninoculated controls. However, 10-strain synthetic microbial communities increased both microbial productivity and duckweed growth more than the average single-strain inoculation and uninoculated controls, meaning that host-microbiome mutualisms can emerge from community interactions among microbes on hosts. The effects of community inoculation were sub-additive, suggesting at least some competition among microbes in the duckweed microbiome. We also investigated the relationship between L. minor fitness and that of its microbes, providing some of the first empirical estimates of broad fitness alignment between plants and members of their microbiomes; hosts grew faster with more productive microbes or microbiomes., Importance: There is currently substantial interest in engineering synthetic microbiomes for health or agricultural applications. One key question is how multi-strain microbial communities differ from single microbial strains in their productivity and effects on hosts. We tested 20 single bacterial strains and 2 distinct 10-strain synthetic communities on plant hosts and found that 10-strain communities led to faster host growth and greater microbial productivity than the average, but not the best, single strain. Furthermore, the microbial strains or communities that achieved the greatest cell densities were also the most beneficial to their hosts, showing that both specific single strains and multi-strain synthetic communities can engage in high-quality mutualisms with their hosts. Our results suggest that ~5% of single strains, as well as multi-strain synthetic communities comprised largely of commensal microbes, can benefit hosts and result in effective host-microbe mutualisms., Competing Interests: The authors declare no conflict of interest.

Microbial-assisted rhizoengineering is a promising biotechnology for improving crop productivity. In this study, lettuce roots were bacterized with two lead (Pb) tolerant rhizobacteria including Pseudomonas azotoformans ESR4 and P. poae ESR6, and a consortium consisted of ESR4 and ESR6 to increase productivity, physiology and antioxidants, and reduce Pb accumulation grown in Pb-contaminated soil i.e., 80 (Pb in native soil), 400 and 800 mg kg -1 Pb. In vitro studies showed that these strains and the consortium produced biofilms, synthesized indole-3-acetic acid and NH 3 , and solubilized phosphate challenging to 0, 100, 200 and 400 mg L -1 of Pb. In static conditions and 400 mg L -1 Pb, ESR4, ESR6 and the consortium adsorbed 317.0, 339.5 and 357.4 mg L -1 Pb, respectively, while 384.7, 380.7 and 373.2 mg L -1 Pb, respectively, in shaking conditions. Fourier transform infrared spectroscopy results revealed that several functional groups [Pb-S, M - O, O-M-O (M = metal ions), S-S, PO, CO, -NH, -NH 2, C-C-O, and C-H] were involved in Pb adsorption. ESR4, ESR6 and the consortium-assisted rhizoengineering (i) increased leaf numbers and biomass production, (ii) reduced H 2 O 2 production, malondialdehyde, electrolyte leakages, and transpiration rate, (iii) augmented photosynthetic pigments, photosynthetic rate, water use efficiency, total antioxidant capacity, total flavonoid content, total phenolic content, and minerals like Ca 2+ and Mg 2+ in comparison to non-rhizoengineering plants grown in Pb-contaminated soil. Principal component analysis revealed that higher pigment production and photosynthetic rate, improved water use efficiency and increased uptake of Ca 2+ were interlinked to increased productivity by bacterial rhizoengineering of lettuce grown in different levels of Pb exposures. Surprisingly, Pb accumulation in lettuce roots and shoots was remarkably decreased by rhizoengineering than in non-rhizoengineering. Thus, these bacterial strains and this consortium could be utilized to improve productivity and reduce Pb accumulation in lettuce., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background and Objective: Quorum sensing (QS) is a chemical language of bacteria, and use of N-acyl-homoserine lactone (AHL) signals is one of the most known mechanisms by which bacteria can communicate with each other to regulate the gene expression based on bacterial density. Previous studies suggested that QS has an important role in pathogenicity of P.aeruginosa, so that the present study investigation the correlation of QS genes in local isolate of P.aeruginosa with production of virulence factors. Materials and Method: The study included collection of 126 samples from various clinical sources and, these samples included wounds, burns, ear, cystic fibrosis and urinary tract infections (UTI). Pyocyanin and biofilm production were determined spectrophotometrically, QS genes were detected by PCR based on specific sequences for lasI and rhlI genes. Results: Final diagnosis showed that 51 isolates belong to target bacteria and, the phenotypic detection of some virulence factors showed that 46 isolates (90%) showed variance in the strength of Pyocyanin production, and 44 (86%) of the isolates showed a differences in their ability to biofilm formation and, screed QS genes showed that 48 (94%) of isolates were positive for lasI gene, while 44 (86%) of isolates have rhlI gene, and this study pointed that 9 isolates have one of QS genes lasI or rhlI, and also have one or more of virulence factors, as well as the results showed that 1 isolate out of 51 negative for both QS genes and weakly in production of virulence factors. Conclusion: Most of local isolates were positive for QS genes and, QS plays an important role in the pathogenesis, not all of the virulence factors controlled by QS. The study indicated that the isolates which have both QS genes were more virulence than isolates that have single gene. [ABSTRACT FROM AUTHOR]

The soil-water interface is replete with photic biofilm and iron minerals; however, the potential of how iron minerals promote biotic nitrate removal is still unknown. This study investigates the physiological and ecological responses of photic biofilm to hematite (Fe 2 O 3 ), in order to explore a practically feasible approach for in-situ nitrate removal. The nitrate removal by photic biofilm was significantly higher in the presence of Fe 2 O 3 (92.5%) compared to the control (82.8%). Results show that the presence of Fe 2 O 3 changed the microbial community composition of the photic biofilm, facilitates the thriving of Magnetospirillum and Pseudomonas, and promotes the growth of photic biofilm represented by the extracellular polymeric substance (EPS) and the content of chlorophyll. The presence of Fe 2 O 3 also induces oxidative stress (•O 2 - ) in the photic biofilm, which was demonstrated by electron spin resonance spectrometry. However, the photic biofilm could improve the EPS productivity to prevent the entrance of Fe 2 O 3 to cells in the biofilm matrix and mitigate oxidative stress. The Fe 2 O 3 then promoted the relative abundance of Magnetospirillum and Pseudomonas and the activity of nitrate reductase, which accelerates nitrate reduction by the photic biofilm. This study provides an insight into the interaction between iron minerals and photic biofilm and demonstrates the possibility of combining biotic and abiotic methods to improve the in-situ nitrate removal rate., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Zinc finger Asp-His-His-Cys motif-containing (zDHHC) proteins, known for their palmitoyltransferase (PAT) activity, play crucial roles in diverse cellular processes, including immune regulation. However, their non-palmitoyltransferase immunomodulatory functions and involvement in teleost immune responses remain underexplored. In this study, we systematically characterized the zDHHC family in the large yellow croaker ( Larimichthys crocea ), identifying 22 members. Phylogenetic analysis unveiled that each of the 22 Lc zDHHCs formed distinct clusters with their orthologues from other teleost species. Furthermore, all Lc zDHHCs exhibited a highly conserved DHHC domain, as confirmed by tertiary structure prediction. Notably, Lc zDHHC23 exhibited the most pronounced upregulation following Pseudomonas plecoglossicida ( P. plecoglossicida ) infection of macrophage/monocyte cells (MO/MΦ). Silencing Lc zDHHC23 led to heightened pro-inflammatory cytokine expression and diminished anti-inflammatory cytokine levels in MO/MΦ during infection, indicating its anti-inflammatory role. Functionally, Lc zDHHC23 facilitated M2-type macrophage polarization, as evidenced by a significant skewing of MO/MΦ towards the pro-inflammatory M1 phenotype upon Lc zDHHC23 knockdown, along with the inhibition of MO/MΦ necroptosis induced by P. plecoglossicida infection. These findings highlight the non-PAT immunomodulatory function of Lc zDHHC23 in teleost immune regulation, broadening our understanding of zDHHC proteins in host-pathogen interactions, suggesting Lc zDHHC23 as a potential therapeutic target for immune modulation in aquatic species., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Dai, Zhao, Zhu, Fei, Nie and Chen.)

In the present study, ten (10) selected bacteria isolated from chasmophytic wild Chenopodium were evaluated for alleviation of drought stress in chickpea. All the bacterial cultures were potential P, K and Zn solubilizer. About 50% of the bacteria could produce Indole-3-acetic acid (IAA) and 1-aminocyclopropane-1-carboxylate (ACC) deaminase. The bacteria showed wide range of tolerance towards pH, salinity, temperature and osmotic stress. Bacillus paralicheniformis L38, Pseudomonas sp. LN75, Enterobacter hormachei subsp. xiangfengensis LJ89, B. paramycoides L17 and Micrococcus luteus LA9 significantly improved growth and nutrient (N, P, K, Fe and Zn) content in chickpea under water stress during a green house experiment conducted following a completely randomized design (CRD). Application of Microbacterium imperiale LJ10, B. stercoris LN74, Pseudomonas sp. LN75, B. paralicheniformis L38 and E. hormachei subsp. xiangfengensis LJ89 reduced the antioxidant enzymes under water stress. During field experiments conducted following randomized block design (RBD), all the bacterial inoculations improved chickpea yield under water stress. Highest yield (1363 kg ha -1 ) was obtained in plants inoculated with Pseudomonas sp. LN75. Pseudomonas sp. LN75, B. paralicheniformis L38 and E. hormachei subsp. xiangfengensis LJ89 have potential as microbial stimulants to alleviate the water stress in chickpea. To the best of our knowledge this is the first report of using chasmophyte associated bacteria for alleviation of water stress in a crop plant., (© 2024. The Author(s).)

Modulating the soil microbiome by applying microbial inoculants has gained increasing attention as eco-friendly option to improve soil disease suppressiveness. Currently, studies unraveling the interplay of inoculants, root-associated microbiome, and plant response are lacking for apple trees. Here, we provide insights into the ability of Bacillus velezensis FZB42 or Pseudomonas sp. RU47 to colonize apple root-associated microhabitats and to modulate their microbiome. We applied the two strains to apple plants grown in soils from the same site either affected by apple replant disease (ARD) or not (grass), screened their establishment by selective plating, and measured phytoalexins in roots 3, 16, and 28 days post inoculation (dpi). Sequencing of 16S rRNA gene and ITS fragments amplified from DNA extracted 28 dpi from different microhabitat samples revealed significant inoculation effects on fungal β-diversity in root-affected soil and rhizoplane. Interestingly, only in ARD soil, most abundant bacterial amplicon sequence variants (ASVs) changed significantly in relative abundance. Relative abundances of ASVs affiliated with Enterobacteriaceae were higher in rhizoplane of apple grown in ARD soil and reduced by both inoculants. Bacterial communities in the root endosphere were not affected by the inoculants but their presence was indicated. Interestingly and previously unobserved, apple plants responded to the inoculants with increased phytoalexin content in roots, more pronounced in grass than ARD soil. Altogether, our results indicate that FZB42 and RU47 were rhizosphere competent, modulated the root-associated microbiome, and were perceived by the apple plants, which could make them interesting candidates for an eco-friendly mitigation strategy of ARD. KEY POINTS: • Rhizosphere competent inoculants modulated the microbiome (mainly fungi) • Inoculants reduced relative abundance of Enterobacteriaceae in the ARD rhizoplane • Inoculants increased phytoalexin content in roots, stronger in grass than ARD soil., (© 2024. The Author(s).)

A common feature of N -acyl-l-homoserine lactone (AHL) quorum-sensing (QS) systems is that the AHL signal is autoinducing. Once induced, a cell will further amplify the signal via a positive feedback loop. Pseudomonas fuscovaginae UPB0736 has two fully functional AHL QS systems, called PfsI/R and PfvI/R, which are inactive in a standard laboratory setting. In this work, we induce the QS systems with exogenous AHL signals and characterize the AHL signal amplification effect and QS activation dynamics at community and single-cell level. While the cognate signal is in both cases significantly further amplified to physiologically relevant levels, we observe only a limited response in terms of AHL synthase gene promoter activity. Additionally, the PfsI/R QS system exhibits a unique dramatic phenotypic heterogeneity, where only up to 5% of all cells amplify the signal further and are, thus, considered to be QS active., Importance: Bacteria use N -acyl-l-homoserine lactone (AHL) quorum-sensing (QS) systems for population-wide phenotypic coordination. The QS configuration in Pseudomonas fuscovaginae is dramatically different from other model examples of AHL QS signaling and, thus, represents an important exception to the norm, which usually states that QS triggers population-wide phenotypic transitions in relation to cell density. We argue that the differences in QS dynamics of P. fuscovaginae highlight its different evolutionary purpose, which is ultimately dictated by the selective pressures of its natural habitat. We hope that this example will further expand our understanding of the complex and yet unknown QS-enabled sociomicrobiology. Furthermore, we argue that exemptions to the QS norm will be found in other plant-pathogenic bacterial strains that grow in similar environments and that molecularly similar QS systems do not necessarily share a similar evolutionary purpose; therefore, generalizations about bacterial cell-to-cell signaling systems function should be avoided., Competing Interests: The authors declare no conflict of interest.

Here, we demonstrate the beneficial effect of surfactant-producing pseudomonads on Pantoea eucalypti 299R. We conducted a series of experiments in environments of increasing complexity. P. eucalypti 299R (Pe299R), and Pseudomonas sp. FF1 (Pff1) or Pe299R and surfactant-production deficient Pseudomonas sp. FF1::ΔviscB (Pff1ΔviscB) were co-inoculated in broth, on swarming agar plates, and on plants. In broth, there were no differences in the growth dynamics of Pe299R when growing in the presence of Pff1 or Pff1ΔviscB. By contrast, on swarming agar plates, Pe299R was able to co-swarm with Pff1 which led to a significant increase in Pe299R biomass compared to Pe299R growing with Pff1ΔviscB or in monoculture. Finally in planta, and using the single-cell bioreporter for reproductive success (CUSPER), we found a temporally distinct beneficial effect of Pff1 on co-inoculated Pe299R subpopulations that did not occur in the presence of Pff1ΔviscB. We tested three additional surfactant-producing pseudomonads and their respective surfactant knockout mutants on PE299R on swarming agar showing similar results. This led us to propose a model for the positive effect of surfactant production during leaf colonization. Our results indicate that co-motility might be common during leaf colonization and adds yet another facet to the already manyfold roles of surfactants., (© 2024. The Author(s).)

Plant-associated microbes play vital roles in promoting plant growth and health, with plants secreting root exudates into the rhizosphere to attract beneficial microbes. Exudate composition defines the nature of microbial recruitment, with different plant species attracting distinct microbiota to enable optimal adaptation to the soil environment. To more closely examine the relationship between plant genotype and microbial recruitment, we analysed the rhizosphere microbiomes of landrace (Chevallier) and modern (NFC Tipple) barley (Hordeum vulgare) cultivars. Distinct differences were observed between the plant-associated microbiomes of the 2 cultivars, with the plant-growth promoting rhizobacterial genus Pseudomonas substantially more abundant in the Tipple rhizosphere. Striking differences were also observed between the phenotypes of recruited Pseudomonas populations, alongside distinct genotypic clustering by cultivar. Cultivar-driven Pseudomonas selection was driven by root exudate composition, with the greater abundance of hexose sugars secreted from Tipple roots attracting microbes better adapted to growth on these metabolites and vice versa. Cultivar-driven selection also operates at the molecular level, with both gene expression and the abundance of ecologically relevant loci differing between Tipple and Chevallier Pseudomonas isolates. Finally, cultivar-driven selection is important for plant health, with both cultivars showing a distinct preference for microbes selected by their genetic siblings in rhizosphere transplantation assays., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Pacheco-Moreno et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The genetic characterization of Pseudomonas protegens Pf-5 was recently completed. However, the inferred metabolic network structure has not yet been evaluated experimentally. Here, we employed 13C-tracers and quantitative flux analysis to investigate the intracellular network for carbohydrate metabolism. In lieu of the direct phosphorylation of glucose by glucose kinase, glucose catabolism was characterized primarily by the oxidation of glucose to gluconate and 2-ketogluconate before the phosphorylation of these metabolites to feed the Entner-Doudoroff (ED) pathway. In the absence of phosphofructokinase activity, a cyclic flux from the ED pathway to the upper Embden-Meyerhof-Parnas (EMP) pathway was responsible for routing glucose-derived carbons to the non-oxidative pentose phosphate (PP) pathway. Consistent with the lack of annotated genes in P. protegens Pf-5 for the transport or initial catabolism of pentoses and galactose, only glucose was assimilated into intracellular metabolites in the presence of xylose, arabinose, or galactose. However, when glucose was fed simultaneously with fructose or mannose, co-uptake of these hexoses was evident, but glucose was preferred over fructose (3 to 1) and over mannose (4 to 1). Despite gene annotation of mannose catabolism to fructose-6-phosphate, metabolite labeling patterns revealed that mannose was assimilated into fructose-1,6-bisphosphate, similarly to fructose catabolism. Remarkably, carbons from mannose and fructose were also found to cycle backward through the upper EMP pathway toward the ED pathway. Therefore, the operational metabolic network for processing carbohydrates in P. protegens Pf-5 prioritizes flux through the ED pathway to channel carbons to EMP, PP, and downstream pathways.IMPORTANCE Species of the Pseudomonas genus thrive in various nutritional environments and have strong biocatalytic potential due to their diverse metabolic capabilities. Carbohydrate substrates are ubiquitous both in environmental matrices and in feedstocks for engineered bioconversion. Here, we investigated the metabolic network for carbohydrate metabolism in Pseudomonas protegens Pf-5. Metabolic flux quantitation revealed the relative involvement of different catabolic routes in channeling carbohydrate carbons through a cyclic metabolic network. We also uncovered that mannose catabolism was similar to fructose catabolism, despite the annotation of a different pathway in the genome. Elucidation of the constitutive metabolic network in P. protegens is important for understanding its innate carbohydrate processing, thus laying the foundation for targeting metabolic engineering of this untapped Pseudomonas species. [ABSTRACT FROM AUTHOR]

Crises are the driving force of the inventions that have led man to overcome himself. The challenge that concerns us today is to maintain our lifestyle with the energy expenditure involved. In the present work, it’s proposed the production of emulsifiers such as monoglycerides (MG) and diglycerides (DG) and biodiesel (FAEE) from renewable sources using Pseudomonas fluorescens lipase immobilized over Ca modified mesoporous SBA-15 supports (L PF /Ca/SBA-15) as biocatalyst. Using a packed bed reactor thermostated at 37 °C, the catalyst was capable to convert sunflower oil and commercial ethanol (96 wt%) into FAEE (35.0 wt%), MG (12.2 wt%) and DG (43.2 wt%). From these results, a separation by vacuum distillation was simulated. A good separation of the biodiesel in the top of the column at approx. 325 °C and the emulsifiers in the bottom of the column at 484 °C, was obtained. Thus, from interdisciplinary technologies such as enzymology, nanostructured materials design and computer simulation, a project for the production and purification of two products, emulsifiers and biodiesel, with one catalyst is offered. [ABSTRACT FROM AUTHOR]

Pseudomonas sp. NCIM 5235 is a caffeine-degrading bacterial strain that metabolizes caffeine by sequential demethylation using methylxanthine demethylases. These enzymes belong to the class of two-component Rieske oxygenases and require an oxidoreductase, NdmD, for efficient catalysis. NdmD in Pseudomonas sp. has a unique domain fusion in its N-terminal that is not observed in any other Rieske oxygenase reductases reported so far. In this report, a ~ 1.7 kb ndmD gene from the gDNA of Pseudomonas sp. has been isolated and has been cloned in a pET28a expression vector. Soluble NdmD was over-expressed in Escherichia coli BL21 cells and purified by Ni2+ NTA chromatography. Monomeric molecular mass of the protein was found to be ~ 65 kDa and optimal activity was observed at 35 °C and pH 8.0. It showed broad substrate specificity with highest Kcat/km of 490.8 ± 17.7 towards cytochrome c. To determine the role of N-terminal Rieske domain in its reductase activity, two deletion constructs Δ114NdmD and Δ250NdmD were made. Cytochrome c reductase (ccr) activity of the NdmD constructs and demethylase activity of NdmA in the presence of NdmD constructs showed that there is no significant difference in the catalytic activity of NdmD upon deletion of its N-terminal Rieske domain. However, there might be some functional and evolutionary significance for the fusion of Rieske domain to NdmD and we hypothesize that this domain fusion is an intermediate phase of evolution towards the development of a more efficient enzyme system for xenobiotic degradation. [ABSTRACT FROM AUTHOR]

Root colonization of plant growth-promoting bacteria is a complex multistep process that is influenced by several factors. For example, during adherence to plant roots, bacteria have to endure reactive oxygen species (ROS) produced by plants. In this study, we report that the global transcriptional regulator Fis is involved in the regulation of ROS-tolerance of Pseudomonas putida and thereby affects barley root colonization. Fis overexpression reduced both ROS-tolerance and adherence to barley roots and activated the transcription of the nuoA-N operon encoding NADH dehydrogenase I, the first enzyme of a membrane-bound electron-transport chain. The nuoA-N knockout mutation in the fis-overexpression background increased the ROS-tolerance and adherence to barley roots. We show that nuoA has two transcriptional start sites located 104 and 377 nucleotides upstream of the coding sequence, indicating the presence of two promoters. The DNase I footprint analysis revealed four Fis binding sites: Fis-nuo1 to Fis-nuo4, situated between these two promoters. Site-directed mutagenesis in a promoter-lacZ reporter and β-galactosidase assay further confirmed direct binding of Fis to Fis-nuo2 and probably to Fis-nuo4 but not to Fis-nuo1 and Fis-nuo3. Additionally, the results implied that Fis binding to Fis-nuo4 could affect transcription of the nuoA-N operon by modification of upstream DNA topology. Moreover, our transposon mutagenesis results indicated that Fis might be involved in the regulation of several alternative ROS detoxification processes utilizing NADH. [ABSTRACT FROM AUTHOR]

Summary: Plants evolved disease resistance (R) proteins that recognize corresponding pathogen effectors and activate effector‐triggered immunity (ETI). However, it is largely unknown why, in some cases, a suppressor of ETI exists in plants. Arabidopsis SOBER1 (Suppressor of AvrBsT‐elicited Resistance 1) was identified previously as a suppressor of Xanthomonas acetyltransferase effector AvrBsT‐triggered immunity. Nevertheless, the extent to which SOBER1 suppresses ETI is unclear. Here, we identified SOBER1 as a suppressor of Pseudomonas acetyltransferase effector HopZ5‐triggered immunity in Arabidopsis using recombinant inbred lines. Further analysis showed that SOBER1 suppresses immunity triggered by multiple bacterial acetyltransferases. Interestingly, SOBER1 interferes with the immunity signalling activated by some but not all tested acetyltransferase effectors, indicating that SOBER1 might target components that are shared between several ETI pathways. [ABSTRACT FROM AUTHOR]

Membrane-bound fatty acid desaturases perform oxygenated desaturation reactions to insert double bonds within fatty acyl chains in regioselective and stereoselective manners. The Δ9-fatty acid desaturase strictly creates the first double bond between C9 and 10 positions of most saturated substrates. As the three-dimensional structures of the bacterial membrane fatty acid desaturases are not available, relevant information about the enzymes are derived from their amino acid sequences, site-directed mutagenesis and domain swapping in similar membrane-bound desaturases. The cold-tolerant Pseudomonas sp. AMS8 was found to produce high amount of monounsaturated fatty acids at low temperature. Subsequently, an active Δ9-fatty acid desaturase was isolated and functionally expressed in Escherichia coli. In this paper we report homology modeling and docking studies of a Δ9-fatty acid desaturase from a Coldtolerant Pseudomonas sp. AMS8 for the first time to the best of our knowledge. Three dimensional structure of the enzyme was built using MODELLER version 9.18 using a suitable template. The protein model contained the three conserved-histidine residues typical for all membrane-bound desaturase catalytic activity. The structure was subjected to energy minimization and checked for correctness using Ramachandran plots and ERRAT, which showed a good quality model of 91.6 and 65.0%, respectively. The protein model was used to preform MD simulation and docking of palmitic acid using CHARMM36 force field in GROMACS Version 5 and Autodock tool Version 4.2, respectively. The docking simulation with the lowest binding energy, 6.8 kcal/mol had a number of residues in close contact with the docked palmitic acid namely, Ile26, Tyr95, Val179, Gly180, Pro64, Glu203, His34, His206, His71, Arg182, Thr85, Lys98 and His177. Interestingly, among the binding residues are His34, His71 and His206 from the first, second, and third conserved histidine motif, respectively, which constitute the active site of the enzyme. The results obtained are in compliance with the in vivo activity of the Δ9-fatty acid desaturase on the membrane phospholipids. [ABSTRACT FROM AUTHOR]

Lipase plays an important role in industrial and biotechnological applications. Lipases have been subject to modification at the N and C terminals, allowing better understanding of lipase stability and the discovery of novel properties. A thermotolerant lipase has been isolated from Antarctic Pseudomonas sp. The purified Antarctic AMS3 lipase (native) was found to be stable across a broad range of temperatures and pHlevels. The lipase has a partial Glutathione-S-transferase type C (GST-C) domain at the N-terminal not found in other lipases. To understand the influence of N-terminal GST-C domain on the biochemical and structural features of the native lipase, the deletion of the GST-C domain was carried out. The truncated protein was successfully expressed in E. coli BL21(DE3). The molecular weight of truncated AMS3 lipase was approximately ~45 kDa. The number of truncated AMS3 lipase purification folds was higher than native lipase. Various mono and divalent metal ions increased the activity of the AMS3 lipase. The truncated AMS3 lipase demonstrated a similarly broad temperature range, with the pH profile exhibiting higher activity under alkaline conditions. The purified lipase showed a substrate preference for a long carbon chain substrate. In addition, the enzyme activity in organic solvents was enhanced, especially for toluene, Dimethylsulfoxide (DMSO), chloroform and xylene. Molecular simulation revealed that the truncated lipase had increased structural compactness and rigidity as compared to native lipase. Removal of the N terminal GST-C generally improved the lipase biochemical characteristics. This enzyme may be utilized for industrial purposes. [ABSTRACT FROM AUTHOR]

Background: Despite the several strategies available for the management of biofilm-associated ventilator-associated pneumonia, data regarding the efficacy of applying antibiotics to the subglottic space (SS) are scarce. We created an in vitro model to assess the efficacy of antibiotic lock therapy (ALT) applied in the SS for eradication of Pseudomonas aeruginosa biofilm in endotracheal tubes (ETTs).Methods: We applied 2 h of ALT to a P. aeruginosa biofilm in ETTs using a single dose (SD) and a 5-day therapy model (5D). We used sterile saline lock therapy (SLT) as the positive control. We compared colony count and the percentage of live cells between both models.Results: The median (IQR) cfu counts/ml and percentage of live cells in the SD-ALT and SD-SLT groups were, respectively, 3.12 × 105 (9.7 × 104-0) vs. 8.16 × 107 (7.0 × 107-0) (p = 0.05) and 53.2% (50.9%-57.2%) vs. 91.5% (87.3%-93.9%) (p < 0.001). The median (IQR) cfu counts/ml and percentage of live cells in the 5D-ALT and 5D-SLT groups were, respectively, 0 (0-0) vs. 3.2 × 107 (2.32 × 107-0) (p = 0.03) and 40.6% (36.6%-60.0%) vs. 90.3% (84.8%-93.9%) (p < 0.001).Conclusion: We demonstrated a statistically significant decrease in the viability of P. aeruginosa biofilm after application of 5D-ALT in the SS. Future clinical studies to assess ALT in patients under mechanical ventilation are needed. [ABSTRACT FROM AUTHOR]

Rhizoctonia solani anastomosis groups (AG)-8 and AG-2-1 and R. oryzae are ubiquitous in cereal-based cropping systems of the Columbia Plateau of the Inland Pacific Northwest and commonly infect wheat. AG-8 and R. oryzae, causal agents of Rhizoctonia root rot and bare patch, are most commonly found in fields in the low-precipitation zone, whereas R. solani AG-2-1 is much less virulent on wheat and is distributed in fields throughout the low-, intermediate-, and high-precipitation zones. Fluorescent Pseudomonas spp. that produce the antibiotic phenazine-1-carboxylic acid (PCA) also are abundant in the rhizosphere of crops grown in the lowprecipitation zone but their broader geographic distribution and elfect on populations of Rhizoctonia is unknown. To address these questions, we surveyed the distribution of PCA producers (Phz+) in 59 fields in cerealbased cropping systems throughout the Columbia Plateau. Phz+ Pseudomonas spp. were detected in 37 of 59 samples and comprised from 0 to 12.5% of the total culturable heterotrophic aerobic rhizosphere bacteria. The frequency with which individual plants were colonized by Phz+ pseudomonads ranged from 0 to 100%. High and moderate colonization frequencies of Phz+ pseudomonads were associated with roots from fields located in the driest areas whereas only moderate and low colonization frequencies were associated with crops where higher annual precipitation occurs. Thus, the geographic distribution of Phz+ pseudomonads overlaps closely with the distribution of R. solani AG-8 but not with that of R. oryzae or R. solani AG-2-1. Moreover, linear regression analysis demonstrated a highly significant inverse relationship between annual precipitation and the frequency of rhizospheres colonized by Phz+ pseudomonads. Phz+ pseudomonads representative of the four major indigenous species (P. aridus, P. cerealis, P. orientalis, and P. synxantha) suppressed Rhizoctonia root rot of wheat when applied as seed treatments. In vitro, mean 50% effective dose values for isolates of AG-8 and AG-2-1 from fields with high and low frequencies of phenazine producers did not differ significantly, nor was there a correlation between virulence of an isolate and sensitivity to PCA, resulting in rejection of the hypothesis that tolerance in Rhizoctonia spp. to PCA develops in nature upon exposure to Phz+ pseudomonads. [ABSTRACT FROM AUTHOR]

Bacterial canker disease caused by Pseudomonas syringae pv. actinidiae, an emerging pathogen of kiwifruit plants, has recently brought about major economic losses worldwide. Genetic studies on virulence functions of P. syringae pv. actinidiae have not yet been reported and there is little experimental data regarding bacterial genes involved in pathogenesis. In this study, we performed a genetic screen in order to identify transposon mutants altered in the lipolytic activity because it is known that mechanisms of regulation, production, and secretion of enzymes often play crucial roles in virulence of plant pathogens. We aimed to identify the set of secretion and global regulatory loci that control lipolytic activity and also play important roles in in planta fitness. Our screen for altered lipolytic activity phenotype identified a total of 58 TnJ transposon mutants. Mapping all these Tn5 mutants revealed that the transposons were inserted in genes that play roles in cell division, chemotaxis, metabolism, movement, recombination, regulation, signal transduction, and transport as well as a few unknown functions. Several of these identified P. syringae pv. actinidiae Tn5 mutants, notably the functions affected in phosphomannomutase AlgC, lipid A biosynthesis acyltransfera.se, glutamate-cysteine ligase, and the type IV pilus protein Pill, were also found affected in in planta survival and/or growth in kiwifruit plants. The results of the genetic screen and identification of novel loci involved in in planta fitness of P. syringae pv. actinidiae are presented and discussed. [ABSTRACT FROM AUTHOR]

It is a well accepted strategy to improve plant salt tolerance through inoculation with beneficial microorganisms. However, its underlying mechanisms still remain unclear. In the present study, hydroponic experiments were conducted to evaluate the effects ofBradyrhizobium japonicumUSDA 110 with salt-tolerantPseudomonas putidaTSAU1 on growth, protein content, nitrogen, and phosphorus uptake as well as root system architecture of soybean (Glycine maxL.) under salt stress. The results indicated that the combined inoculation with USDA 110 and TSAU1 significantly improved plant growth, nitrogen and phosphorus contents, and contents of soluble leaf proteins under salt stress compared to the inoculation with the symbiont alone or compared to un-inoculated ones. The root architectural traits, like root length, surface area, project area, and root volume; as well as nodulation traits were also significantly increased by co-inoculation with USDA 110 and TSAU1. The plant-growth promoting rhizobacteria (PGPR)P. putidastrain TSAU1 could improve the symbiotic interaction between the salt-stressed soybean andB. japonicumUSDA 110. In conclusion, inoculation withB. japonicumand salt-tolerantP. putidasynergistically improved soybean salt tolerance through altering root system architecture facilitating nitrogen and phosphorus acquisition, and nodule formation. [ABSTRACT FROM AUTHOR]

Procedure of manufacturing K. pinnata water extracts containing cecropin P1 (CecP1) from the formerly described transgenic plants is established. It included incubation of leaves at +4°C for 7 days, mechanical homogenization of leaves using water as extraction solvent, and heating at +70°C for inactivating plant enzymes. Yield of CecP1 (after heating and sterilizing filtration) was 0.3% of total protein in the extract. The water extract of K. pinnata + CecP1 exhibits favorable effect on healing of wounds infected with S. aureus (equal to Cefazolin) and with a combination of S. aureus with P. aeruginosa (better than Cefazolin). Wild-type K. pinnata extract exhibited evident microbicide activity against S. aureus with P. aeruginosa but it was substantially strengthened in K. pinnata + CecP1 extract. K. pinnata extracts (both wild-type and transgenic) did not exhibit general toxicity and accelerated wound recovery. Due to immunomodulating activity, wild-type K. pinnata extract accelerated granulation of the wound bed and marginal epithelialization even better than K. pinnata + CecP1 extract. Immunomodulating and microbicide activity of K. pinnata synergizes with microbicide activity of CecP1 accelerating elimination of bacteria. [ABSTRACT FROM AUTHOR]

Expression of heat shock proteins (Hsps) synthesized by bacterium (Pseudomonas spp.) during stress condition (elevated temperature) is identified and studied their association in adaptation to environmental extremes. Collection, screening, isolation and identification of zoom land soil bacterium were carried out by growing bacterial cells on selective Agar Cetrimide media. Optimization of culture conditions (incubation time and temperature) for the maximum production of cell (Dry Cell Weight) also considered for the identified organism (Pseudomonas spp.) in LB media during micro-aerobic batch culture; where maximum cell growth was found 1.80 gm/L at 10 hours incubation period and 37°C incubation temperature, while rpm was 120. Subsequently, bacterial cell was grown in LB media for 6 hours at 37°C and 120 rpm into shaking incubator, then incubated for 2 hours, where maximum cell growth was observed 2.38 ± 0.06 gm/L during elevated temperature (50°C), stimulated by heat shock proteins. Then different intracellular and extracellular soluble protein extraction, purification was attempted from soil bacterium by centrifugation, sonication, repeated freezing and thawing. SDS-PAGE was performed for the separation of heat shock protein, where protein bands intensity 90, 84, 70 and 45 kDa were found according to their molecular weight. Pseudomonas spp. can produce heat shock protein during harsh condition which can help them to survive by effective adaptation mediated by universal regulatory mechanisms affecting several pathways. Hsps are expected to play a significant role to conferring tolerance in response to changes in high temperature. They are also responsible for survival of bacterial cells and increasing soil fertility which associated with virulence. So, this study will help to understand the impact of soil microorganisms to revive the soil fertility though people burn their paddy fields after harvesting in zoom areas of Bangladesh. [ABSTRACT FROM AUTHOR]

The aim of this study was to isolate and select lipase producing bacteria from effluent waste of paint industry. Five lipase producing Gram negative bacterial isolates showed promising extracellular lipase activity. Molecular identification on the basis of 16S rDNA identified bacterial isolates as Aeromonas sp. Sh 2 (KT186103.1), Aeromonas sp. Sh 8 (KT229540.1), Aeromonas sp. Sh 12 (KT427918.1), Pseudomonas sp. Sh 13 (KT251204.1), Pseudomonas sp. Sh 17 (KT251205.1). Phylogenetic analysis revealed that all the three Aeromonas spp. and two Pseudomonas spp. are highly divergent within each group. Effect of various physio-chemical parameters showed that pH 7.0 and temperature 37∘C were favourable for lipase production in all five isolates. Glucose in combination with peptone enhanced the lipase activity from 1359 to 2347 Umg-1 in Aeromonas sp. Sh 2. Combination of glucose with yeast extract improved the lipase activity of Aeromonas sp. Sh 8 from 1641 to 2674 Umg-1, whereas glucose with casein hydrolysate increased the lipase activity of Aeromonas sp. Sh 12 from 1568 to 1983 Umg-1. Pseudomonas sp. Sh 13 and Pseudomonas sp. Sh 17 did not show enhanced lipase activity in presence of different carbon or nitrogen sources. These bacterial isolates could be exploited to treat oil and paint based industrial effluents and contribute to reduce environmental hazards. [ABSTRACT FROM AUTHOR]

Background: Pandoraea spp. are recently discovered bacteria, mainly recovered from cystic fibrosis (CF) patients, but their epidemiology and clinical significance are not well known. We describe an epidemic spread of Pandoraea pulmonicola from 2009 in our CF center, involving 6 out of 243 CF patients.Methods: Bacterial identification used amplified ribosomal DNA restriction analysis (ARDRA), MALDI-TOF mass spectrometry (MALDI-TOF MS) and 16S rDNA gene sequencing. The clonal link between strains was assessed with pulsed field gel electrophoresis (PFGE) using XbaI. Clinical data were gathered for all patients.Results: The index case was chronically colonized since 2000. The main hypothesis for this bacterial spread was a droplet cross-transmission, due to preventive measures not being strictly followed. Antibiotic susceptibility testing revealed resistance to beta-lactams, ciprofloxacin and colistin. However, there was susceptibility to trimethoprim-sulfamethoxazole. All patients were chronically colonized with Pseudomonas aeruginosa, and the acquisition of P. pulmonicola resulted in chronic colonization in all patients. Three patients died, and two patients remained clinically stable, whereas one patient had a decline in lung function.Conclusions: This study, which is the first to describe an epidemic spread of P. pulmonicola, notes the potential transmissibility of this bacterial species and the need for infection control measures. [ABSTRACT FROM AUTHOR]

Variability in whole-cell biocatalyst performance represents a critical aspect for stable and productive bioprocessing. In order to investigate whether and how oxygenase-catalyzed reactions are affected by such variability issues in solvent-tolerant Pseudomonas, different inducers, expression systems, and host strains were tested for the reproducibility of xylene and styrene monooxygenase catalyzed hydroxylation and epoxidation reactions, respectively. Significantly higher activity variations were found for biocatalysts based on solvent-tolerant Pseudomonas putida DOT-TIE and S12 compared with solvent-sensitive P. putida KT2440, Escherichia coli JM101, and solvent-tolerant Pseudomonas taiwanensis VLB120. Specific styrene epoxidation rates corresponded to cellular styrene monooxygenase contents. Detected variations in activity strictly depended on the type of regulatory system employed, being high with the alk- and low with the lac-system. These results show that the occurrence of clonal variability in recombinant gene expression in Pseudomonas depends on the combination of regulatory system and host strain, does not correlate with a general phenotype such as solvent tolerance, and must be evaluated case by case. [ABSTRACT FROM AUTHOR]

High temperature (HT), high humidity (HH), and pathogen infection often co-occur and negatively affect plant growth. However, these stress factors and plant responses are generally studied in isolation. The mechanisms of synergistic responses to combined stresses are poorly understood. We isolated the subgroup IIb WRKY family member CaWRKY6 from Capsicum annuum and performed quantitative real-time PCR analysis. CaWRKY6 expression was upregulated by individual or simultaneous treatment with HT, HH, combined HT and HH (HTHH), and Ralstonia solanacearum inoculation, and responded to exogenous application of jasmonic acid (JA), ethephon, and abscisic acid (ABA). Virus-induced gene silencing of CaWRKY6 enhanced pepper plant susceptibility to R. solanacearum and HTHH, and downregulated the hypersensitive response (HR), JA-, ethylene (ET)-, and ABA-induced marker gene expression, and thermotolerance-associated expression of CaHSP24, ER-small CaSHP, and Chl-small CaHSP. CaWRKY6 overexpression in pepper attenuated the HTHH-induced suppression of resistance to R. solanacearum infection. CaWRKY6 bound to and activated the CaWRKY40 promoter in planta, which is a pepper WRKY that regulates heatstress tolerance and R. solanacearum resistance. CaWRKY40 silencing significantly blocked HR-induced cell death and reduced transcriptional expression of CaWRKY40. These data suggest that CaWRKY6 is a positive regulator of R. solanacearum resistance and heat-stress tolerance, which occurs in part by activating CaWRKY40. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa is an opportunistic pathogen of plants and animals, which produces virulence factors in order to infect or colonize its eukaryotic hosts. Cyclodipeptides (CDPs) produced by P. aeruginosa exhibit cytotoxic properties toward human tumor cells. In this study, we evaluated the effect of a CDP mix, comprised of cyclo(L-Pro-L-Tyr), cyclo(L-Pro-L-Val), and cyclo(L-Pro-L-Phe) that were isolated from P. aeruginosa, on two human cancer cell lines. Our results demonstrated that the CDP mix promoted cell death in cultures of the HeLa cervical adenocarcinoma and Caco-2 colorectal adenocarcinoma cell lines in a dose-dependent manner, with a 50% inhibitory concentration (IC50) of 0.53 and 0.66 mg/mL, for HeLa and Caco-2 cells, respectively. Flow cytometric analysis, using annexin V and propidium iodide as apoptosis and necrosis indicators, respectively, clearly showed that HeLa and Caco-2 cells exhibited apoptotic characteristics when treated with the CDP mix at a concentration <0.001 mg/mL. IC50 values for apoptotic cells in HeLa and Caco-2 cells were 6.5 × 10−5 and 1.8 × 10−4 mg/mL, respectively. Our results indicate that an apoptotic pathway is involved in the inhibition of cell proliferation caused by the P. aeruginosa CDP mix. [ABSTRACT FROM AUTHOR]

The performances of betaine on the metabolic processes of vitamin B-producing Pseudomonas denitrificans were investigated in this paper. The results showed that betaine was an indispensable methyl-group donor for vitamin B biosynthesis, but large amounts of the extracellular glycine accompanied by betaine metabolism would impose a severe restriction on the cell growth of P. denitrificans. By further using a comparative metabolomics approach coupled with intracellular free amino acids analysis for the fermentation processes with betaine addition (10 g/l) or not, it was found that betaine could highly strengthen the formation of some key precursors and intermediates facilitating vitamin B biosynthesis, such as δ-aminolevulinic acid (ALA, the first precursor of vitamin B), glutamate (an intermediate of ALA via C pathway), glycine (an intermediate of ALA via C pathway), and methionine (directly participating in the methylation reaction involved in vitamin B biosynthetic pathway). Therefore, the performances of betaine on P. denitrificans metabolic processes were not only serving as a decisive methyl-group donor for vitamin B biosynthesis, but also playing a powerfully promoting role in the generation of vitamin B precursors and intermediates. [ABSTRACT FROM AUTHOR]

Two polyhydroxyalkanoate depolymerases, PHAase I and PHAase II, were purified to homogeneity from the culture supernatant of an effective PHA-degrading bacterium, Pseudomonas mendocina DS04-T. The molecular masses of PHAase I and PHAase II were determined by SDS-PAGE as 59.4 and 33.8 kDa, respectively. Their optimum pH values were 8.5 and 8, respectively. Enzymatic activity was optimal at 50 °C. Both purified enzymes could degrade PHB, PHBV, and P(3HB-co-4HB). Addition of Na and K slightly increased the rate of PHAase II. EDTA significantly inhibited PHAase II but not PHAase I. Mercaptoethanol and HO also inhibited the activities of both enzymes. [ABSTRACT FROM AUTHOR]

Accurate detection of subpopulation size determinations in bimodal populations remains problematic yet it represents a powerful way by which cellular heterogeneity under different environmental conditions can be compared. So far, most studies have relied on qualitative descriptions of population distribution patterns, on population-independent descriptors, or on arbitrary placement of thresholds distinguishing biological ON from OFF states. We found that all these methods fall short of accurately describing small population sizes in bimodal populations. Here we propose a simple, statistics-based method for the analysis of small subpopulation sizes for use in the free software environment R and test this method on real as well as simulated data. Four so-called population splitting methods were designed with different algorithms that can estimate subpopulation sizes from bimodal populations. All four methods proved more precise than previously used methods when analyzing subpopulation sizes of transfer competent cells arising in populations of the bacterium Pseudomonas knackmussii B13. The methods’ resolving powers were further explored by bootstrapping and simulations. Two of the methods were not severely limited by the proportions of subpopulations they could estimate correctly, but the two others only allowed accurate subpopulation quantification when this amounted to less than 25% of the total population. In contrast, only one method was still sufficiently accurate with subpopulations smaller than 1% of the total population. This study proposes a number of rational approximations to quantifying small subpopulations and offers an easy-to-use protocol for their implementation in the open source statistical software environment R. [ABSTRACT FROM AUTHOR]

Haloarchaeal diversity of Inche-Boroun wetland in north of Iran in Golestan province was investigated by using culture-dependent methods. Sampling was carried out in May and September 2010. In each sampling, 4 distinct region of wetland were analyzed by using complex media like MGM, JCM168, MH1 and an alkaliphilic medium containing 23% salts. After incubation at 40ºC, a total of 406 isolates were prepared and 2.1 x 106 CFU/ml were obtained in culture media. Among all isolates, 361 isolates were obtained from MGM and 39 isolates from JCM 168, 3 isolates from MH1 and 3 isolate from alkaliphilic media. Initial morphological, biochemical and physiological tests were performed. According to the results, 45 isolates were selected and phylogenetic analysis of 16S rRNA was performed for them. Among selected strains, 40 isolates belonged to Halobacteriacaea and were related to Haloarcula, Halorubrum, Haloferax, Halobellus, Halogeometricum, Halobacterium, Halolamina, Halorhabdus and Halostagnicola (respectively 30, 27.5, 17.5, 10, 5.2, 2.6, 2.6, 2.6 and 2.6 percent of Haloarchaeal strains). A total of 5 strains belonged to the kingdom of Bacteria and were related to Rhodovibrio, Pseudomonas and Salicola (respectively 40, 40 and 20 percent of bacterial strains). According to our results and the limited numbers of haloarchaeal genera that having been discovered until now, it seemed that the culturable prokaryotic populations in this hypersaline environment was diverse. [ABSTRACT FROM AUTHOR]

Microbial physiological responses resulting from inappropriate bioprocessing conditions may have a marked impact on process performance within any fermentation system. The influence of different pH-control strategies on physiological status, microbial growth and lactobionic acid production from whey by Pseudomonas taetrolens during bioreactor cultivations has been investigated for the first time in this work. Both cellular behaviour and bioconversion efficiency from P. taetrolens were found to be negatively influenced by pH-control modes carried out at values lower than 6.0 and higher than 7.0. Production schemes were also influenced by the operational pH employed, with asynchronous production from damaged and metabolically active subpopulations at pH values lower than 6.0. Moreover, P. taetrolens showed reduced cellular proliferation and a subsequent delay in the onset of the production phase under acidic conditions (pH < 6.0). Unlike cultivations performed at 6.5, both pH-shift and pH-stat cultivation strategies performed at pH values lower than 6.0 resulted in decreased lactobionic acid production. Whereas the cellular response showed a stress-induced physiological response under acidic conditions, healthy functional cells were predominant at medium operational pH values (6.5-7.0). P. taetrolens thus displayed a robust physiological status at initial pH value of 6.5, resulting in an enhanced bioconversion yield and lactobionic acid productivity (7- and 4-fold higher compared to those attained at initial pH values of 4.5 and 5.0, respectively). These results have shown that pH-control modes strongly affected both the physiological response of cells and the biological performance of P. taetrolens, providing key information for bio-production of lactobionic acid on an industrial scale. [ABSTRACT FROM AUTHOR]

As a highly infectious epidemic in aquaculture, Pseudomonas plecoglossicida infection results in high mortality of teleosts and serious economic losses. Host-pathogen interactions shape the outcome of an infection, yet we still understand little about the molecular mechanism of these pathogen-mediated processes. Here, a P. plecoglossicida strain (NZBD9) and Epinephelus coioides were investigated as a model system to characterize pathogen-induced host metabolic remodeling over the course of infection. We present a non-targeted metabolomics profiling of E. coioides spleens from uninfected E. coioides and those infected with wild-type and clpV-RNA interference (RNAi) strains. The most significant changes of E. coioides upon infection were associated with amino acids, lysophospatidylcholines, and unsaturated fatty acids, involving disturbances in host nutritional utilization and immune responses. Dihydrosphingosine and fatty acid 16:2 were screened as potential biomarkers for assessing P. plecoglossicida infection. The silencing of the P. plecoglossicida clpV gene significantly recovered the lipid metabolism of infected E. coioides. This comprehensive metabolomics study provides novel insights into how P. plecoglossicida shape host metabolism to support their survival and replication and highlights the potential of the virulence gene clpV in the treatment of P. plecoglossicida infection in aquaculture., (© 2022. The Author(s).)

Pseudomonas aeruginosa and other Gram-negative bacteria release membrane vesicles (MVs) from their surfaces, and MVs have an ability to interact with bacterial cells. Although it has been known that many bacteria have mechanisms that control their phenotypes with the transition from exponential phase to stationary phase, changes of properties in released MVs have been poorly understood. Here, we demonstrate that MVs released by P. aeruginosa during the exponential and stationary phases possess different physiochemical properties. MVs purified from the stationary phase had higher buoyant densities than did those purified from the exponential phase. Surface charge, characterized by zeta potential, of MVs tended to be more negative as the growth shifted to the stationary phase, although the charges of PAO1 cells were not altered. Pseudomonas quinolone signal (PQS), one of the regulators related to MV production in P. aeruginosa, was lower in MVs purified from the exponential phase than in those from the stationary phase. MVs from the stationary phase more strongly associated with P. aeruginosa cells than did those from the exponential phase. Our findings suggest that properties of MVs are altered to readily interact with bacterial cells along with the growth transition in P. aeruginosa. [ABSTRACT FROM AUTHOR]

In the camphor monooxygenase system from Pseudomonas putida, the [2Fe-2S]-containing putidaredoxin (Pdx) shuttles electrons between the NADH-dependent putidaredoxin reductase (Pdr) and cytochrome P450cam The mechanism of the PdrPdx redox couple has been investigated by a variety of techniques. One of the exceptions is x-ray crystallography as the native partners associate weakly and resist co-crystallization. Here, we present the 2.6-Å x-ray structure of a catalytically active complex between Pdr and Pdx C73S1C85S chemically cross-linked via the Lys409Pdr.Glu72Pdx pair. The 365 Å2 Pdr-Pdx interface is predominantly hydrophobic with one central Arg310Pdr-Asp38Pdx salt bridge, likely assisting docking and orienting the partners optimally for electron transfer, and a few peripheral hydrogen bonds. A predicted 12-Å-long electron transfer route between FAD and [2Fe-2S] includes flavin flanking Trp330Pdr and the iron ligand Cys39Pdx. The x-ray model agrees well with the experimental and theoretical results and suggests that the linked Pdx must undergo complex movements during turnover to accommodate P450cam; which could limit the Pdx-to-P450cam electron transfer reaction. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa is a gram-negative pathogenic bacterium with a high adaptive potential that allows proliferation in a broad range of hosts or niches. It is also the causative agent of both acute and chronic biofilm-related infections in humans. Three cup gene clusters (cupA-C), involved in the assembly of cell surface fimbriae, have been shown to be involved in biofilm formation by the P. aeruginosa strains PAO1 or PAK. In PA14 isolates, a fourth cluster, named cupD, was identified within a pathogenicity island, PAPI-I, and may contribute to the higher virulence of this strain. Expression of the cupA genes is controlled by the HNS-like protein MvaT, whereas the cupB and cupC genes are under the control of the RocS1A1R twocomponent system. In this study, we show that cupD gene expression is positively controlled by the response regulator RcsB. As a consequence, CupD fimbriae are assembled on the cell surface, which results in a number of phenotypes such as a small colony morphotype, increased biofilm formation and decreased motility. These behaviors are compatible with the sessile bacterial lifestyle. The balance between planktonic and sessile lifestyles is known to be linked to the intracellular levels of c-di-GMP with high levels favoring biofilm formation. We showed that the EAL domain-containing PvrR response regulator counteracts the activity of RcsB on cupD gene expression. The action of PvrR is likely to involve c-di-GMP degradation through phosphodiesterase activity, confirming the key role of this second messenger in the balance between bacterial lifestyles. The regulatory network between RcsB and PvrR remains to be elucidated, but it stands as a potential model system to study how the equilibrium between the two lifestyles could be influenced by therapeutic agents that favor the planktonic lifestyle. This would render the pathogen accessible for the immune system or conventional antibiotic treatment. [ABSTRACT FROM AUTHOR]

Abstract: In order to clarify the physiological role of the merE gene of transposon Tn21, a pE4 plasmid that contained the merR gene of plasmid pMR26 from Pseudomonas strain K-62, and the merE gene of Tn21 from the Shigella flexneri plasmid NR1 (R100) was constructed. Bacteria with plasmid pE4 (merR-o/p-merE) were more hypersensitive to CH3Hg(I) and Hg(II), and took up significantly more CH3Hg(I) and Hg(II), than the isogenic strain. The MerE protein encoded by pE4 was localized in the membrane cell fraction, but not in the soluble fraction. Based on these experimental results, we suggest for the first time that the merE gene is a broad mercury transporter mediating the transport of both CH3Hg(I) and Hg(II) across the bacterial membrane. [Copyright &y& Elsevier]

The article focuses on the ice nucleation activity (INA) of Pseudomonas borealis. INA has been viewed to be an important factor of the crystallization of water. Ice nucleators can be found in bacteria which have been speculated to be a contributor to Pseudomonas borealis' pathogenicity. The discovery of bacteria in melted water has been used to analyze the ice association and gene sequence. The bacterial strains, the characterization of P. borealis, and ice nucleation assays and ice nucleation protein (INP) sequence determination procedures are also examined.