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1.
Curr Microbiol ; 81(7): 184, 2024 May 21.
Article in English | MEDLINE | ID: mdl-38771325

ABSTRACT

Agriculture and livestock management practices known as organic farming rely more on internal processes than external inputs. Natural environments depend heavily on diversity, and organic farming incorporates both the stated purpose of fostering diversity as well as the use of diversity as a management tool. A more complete understanding of agriculture in terms of agro-ecology has begun to be questioned by the traditional reductionist approach to the study of agriculture. Therefore it is necessary to be aware more about the significance of microbes in processes including soil growth, plant nourishment, and the eradication of plant disease, pest, and weeds. In this study, fluorescent Pseudomonas strain (EFP56) and Trichoderma harzianum were studied for antifungal and antibacterial activity against four common root rot fungi and four common laboratory bacteria in vitro experiments. Furthermore, soil-borne disease surveillance and nutritional quality of Lagenaria siceraria, fluorescent Pseudomonas strain (EFP56) and Trichoderma harzianum were combined with neem cake and cotton cake to check their efficacy. Through the application of organic soil amendments in combination with biocontrol agents improved the quality of vegetables and their nutritional value by raising their polyphenol, carbohydrate, and protein content as well as enhancing antioxidant scavenging status. The experiments were conducted in pots and in fields to confirm their efficacy rate. The final outcomes also revealed greater induction of defense system, disease lessening and enriched fruit quality. Consortium of neem cake and cotton cake with bio-stimulants can regulate biotic as well as abiotic stress.


Subject(s)
Endophytes , Pseudomonas , Soil Microbiology , Endophytes/physiology , Pseudomonas/physiology , Cucurbitaceae/microbiology , Plant Diseases/microbiology , Plant Diseases/prevention & control , Hypocreales/physiology , Fungi/physiology , Fungi/drug effects , Bacteria/classification , Bacteria/drug effects , Biological Control Agents , Plant Roots/microbiology , Antifungal Agents/pharmacology , Antifungal Agents/metabolism
2.
Chemosphere ; 358: 142136, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38692363

ABSTRACT

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 (Fe2O3), 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 Fe2O3 (92.5%) compared to the control (82.8%). Results show that the presence of Fe2O3 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 Fe2O3 also induces oxidative stress (•O2-) 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 Fe2O3 to cells in the biofilm matrix and mitigate oxidative stress. The Fe2O3 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.


Subject(s)
Biofilms , Ferric Compounds , Nitrates , Ferric Compounds/metabolism , Ferric Compounds/chemistry , Nitrates/metabolism , Oxidative Stress , Pseudomonas/physiology , Pseudomonas/metabolism
3.
J Appl Microbiol ; 135(5)2024 May 01.
Article in English | MEDLINE | ID: mdl-38632051

ABSTRACT

AIMS: We aimed to develop an effective bacterial combination that can combat Fusarium oxysporum infection in watermelon using in vitro and pot experiments. METHODS AND RESULTS: In total, 53 strains of Bacillus and 4 strains of Pseudomonas were screened. Pseudomonas strains P3 and P4 and Bacillus strains XY-2-3, XY-13, and GJ-1-15 exhibited good antagonistic effects against F. oxysporum. P3 and P4 were identified as Pseudomonas chlororaphis and Pseudomonas fluorescens, respectively. XY-2-3 and GJ-1-15 were identified as B. velezensis, and XY-13 was identified as Bacillus amyloliquefaciens. The three Bacillus strains were antifungal, promoted the growth of watermelon seedlings and had genes to synthesize antagonistic metabolites such as bacilysin, surfactin, yndj, fengycin, iturin, and bacillomycin D. Combinations of Bacillus and Pseudomonas strains, namely, XY-2-3 + P4, GJ-1-15 + P4, XY-13 + P3, and XY-13 + P4, exhibited a good compatibility. These four combinations exhibited antagonistic effects against 11 pathogenic fungi, including various strains of F. oxysporum, Fusarium solani, and Rhizoctonia. Inoculation of these bacterial combinations significantly reduced the incidence of Fusarium wilt in watermelon, promoted plant growth, and improved soil nutrient availability. XY-13 + P4 was the most effective combination against Fusarium wilt in watermelon with the inhibition rate of 78.17%. The number of leaves; aboveground fresh and dry weights; chlorophyll, soil total nitrogen, and soil available phosphorus content increased by 26.8%, 72.12%, 60.47%, 16.97%, 20.16%, and 16.50%, respectively, after XY-13 + P4 inoculation compared with the uninoculated control. Moreover, total root length, root surface area, and root volume of watermelon seedlings were the highest after XY-13 + P3 inoculation, exhibiting increases by 265.83%, 316.79%, and 390.99%, respectively, compared with the uninoculated control. CONCLUSIONS: XY-13 + P4 was the best bacterial combination for controlling Fusarium wilt in watermelon, promoting the growth of watermelon seedlings, and improving soil nutrient availability.


Subject(s)
Bacillus , Citrullus , Disease Resistance , Fusarium , Plant Diseases , Pseudomonas , Fusarium/growth & development , Citrullus/microbiology , Citrullus/growth & development , Plant Diseases/microbiology , Plant Diseases/prevention & control , Bacillus/physiology , Bacillus/genetics , Bacillus/growth & development , Pseudomonas/growth & development , Pseudomonas/physiology , Antibiosis , Pseudomonas fluorescens/growth & development , Seedlings/growth & development , Seedlings/microbiology , Antifungal Agents/pharmacology
4.
Fish Shellfish Immunol ; 149: 109584, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38670411

ABSTRACT

Pseudomonas plecoglossicida, the causative agent of Visceral White Spot Disease, poses substantial risks to large yellow croaker (Larimichthys crocea) aquaculture. Previous genome-wide association studies (GWAS), directed towards elucidating the resistance mechanisms of large yellow croaker against this affliction, suggested that the transmembrane protein 208 (named Lctmem208) may confer a potential advantage. TMEM proteins, particularly TMEM208 located in the endoplasmic reticulum, plays significant roles in autophagy, ER stress, and dynamics of cancer cell. However, research on TMEM's function in teleost fish immunity remains sparse, highlighting a need for further study. This study embarks on a comprehensive examination of LcTmem208, encompassing cloning, molecular characterization, and its dynamics in immune function in response to Pseudomonas plecoglossicida infection. Our findings reveal that LcTmem208 is highly conserved across teleost species, exhibiting pronounced expression in immune-relevant tissues, which escalates significantly upon pathogenic challenge. Transcriptome analysis subsequent to LcTmem208 overexpression in kidney cells unveiled its pivotal role in modulating immune-responsive processes, notably the p53 signaling pathway and cytokine-mediated interactions. Enhanced phagocytic activity in macrophages overexpressing LcTmem208 underscores its importance in innate immunity. Taken together, this is the first time reported the critical involvement of LcTmem208 in regulating innate immune responses of defensing P. plecoglossicida, thereby offering valuable insights into teleost fish immunity and potential strategies for the selective breeding of disease-resistant strains of large yellow croaker in aquaculture practices.


Subject(s)
Fish Diseases , Fish Proteins , Gene Expression Profiling , Immunity, Innate , Perciformes , Pseudomonas Infections , Pseudomonas , Animals , Fish Diseases/immunology , Perciformes/immunology , Perciformes/genetics , Fish Proteins/genetics , Fish Proteins/immunology , Pseudomonas/physiology , Immunity, Innate/genetics , Gene Expression Profiling/veterinary , Pseudomonas Infections/immunology , Pseudomonas Infections/veterinary , Gene Expression Regulation/immunology , Membrane Proteins/genetics , Membrane Proteins/immunology , Transcriptome , Phylogeny , Sequence Alignment/veterinary , Cloning, Molecular
5.
PLoS Biol ; 22(4): e3002232, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38662644

ABSTRACT

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.


Subject(s)
Genotype , Hordeum , Microbiota , Plant Roots , Pseudomonas , Rhizosphere , Hordeum/microbiology , Hordeum/genetics , Hordeum/metabolism , Plant Roots/microbiology , Plant Roots/metabolism , Microbiota/physiology , Microbiota/genetics , Pseudomonas/genetics , Pseudomonas/metabolism , Pseudomonas/physiology , Soil Microbiology , Plant Exudates/metabolism
6.
Plant Dis ; 108(4): 996-1004, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38613135

ABSTRACT

Bacterial wilt caused by Ralstonia solanacearum (RS) is one of the most devastating diseases in patchouli (Pogostemon cablin [Blanco] Benth.), which results in low yield and quality of patchouli. However, no stable and effective control methods have been developed yet. To evaluate the potential of dominant bacterial endophytes in biocontrol, the endophytic bacterial diversity of patchouli was investigated based on Illumina sequencing analysis, and the ability of isolates belonging to the dominant bacterial genera to control RS wilt of patchouli was explored in pot experiments. A total of 245 bacterial genera were detected in patchouli plants, with the highest relative abundance of operational taxonomic units belonging to the genus Pseudomonas detected in roots, leaves, and stems. The Pseudomonas isolates S02, S09, and S26 showed antagonistic activity against RS in vitro and displayed many plant growth-promoting characteristics, including production of indole-3-acetic acid, siderophores, and 1-aminocyclopropane-1-carboxylic acid deaminase and phosphate- and potassium-solubilizing capability. Inoculation of patchouli plants with the isolates S02, S09, and S26 significantly improved shoot growth and decreased the incidence of bacterial wilt caused by RS. The results suggest that screening of dominant bacterial endophytes for effective biocontrol agents based on Illumina sequencing analysis is more efficient than random isolation and screening procedures.


Subject(s)
Endophytes , Plant Diseases , Ralstonia solanacearum , Ralstonia solanacearum/physiology , Ralstonia solanacearum/genetics , Plant Diseases/microbiology , Plant Diseases/prevention & control , Endophytes/genetics , Endophytes/physiology , Endophytes/isolation & purification , Pseudomonas/genetics , Pseudomonas/physiology , High-Throughput Nucleotide Sequencing , Phylogeny , Biological Control Agents
7.
Microb Ecol ; 87(1): 62, 2024 Apr 29.
Article in English | MEDLINE | ID: mdl-38683223

ABSTRACT

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.


Subject(s)
Pantoea , Pseudomonas , Surface-Active Agents , Pantoea/genetics , Pantoea/metabolism , Pantoea/physiology , Pantoea/growth & development , Pseudomonas/metabolism , Pseudomonas/genetics , Pseudomonas/growth & development , Pseudomonas/physiology , Surface-Active Agents/metabolism
8.
Microbiol Spectr ; 12(5): e0417923, 2024 May 02.
Article in English | MEDLINE | ID: mdl-38511955

ABSTRACT

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.


Subject(s)
Acyl-Butyrolactones , Ligases , Pseudomonas , Quorum Sensing , Pseudomonas/genetics , Pseudomonas/physiology , Acyl-Butyrolactones/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Gene Expression Regulation, Bacterial , Promoter Regions, Genetic
9.
Plant Physiol Biochem ; 203: 108080, 2023 Oct.
Article in English | MEDLINE | ID: mdl-37812990

ABSTRACT

Although amelioration of drought stress by Plant Growth Promoting Rhizobacteria (PGPR) is a well-documented phenomenon, the combined molecular and metabolic mechanisms governing this process remain unclear. In these lines, the present study aimed to provide new insights in the underlying drought attenuating mechanisms of tomato plants inoculated with a PGP Pseudomonas putida strain, by using a combination of metabolomic and transcriptomic approaches. Following Differentially Expressed Gene analysis, it became evident that inoculation resulted in a less disturbed plant transcriptome upon drought stress. Untargeted metabolomics highlighted the differential metabolite accumulation upon inoculation, as well as the less metabolic reprograming and the lower accumulation of stress-related metabolites for inoculated stressed plants. These findings were in line with morpho-physiological evidence of drought stress mitigation in the inoculated plants. The redox state modulation, the more efficient nitrogen assimilation, as well as the differential changes in amino acid metabolism, and the induction of the phenylpropanoid biosynthesis pathway, were the main drought-attenuating mechanisms in the SAESo11-inoculated plants. Shifts in pathways related to hormonal signaling were also evident upon inoculation at a transcript level and in conjunction with carbon metabolism regulation, possibly contributed to a drought-attenuation preconditioning. The identified signatory molecules of SAESo11-mediated priming against drought included aspartate, myo-inositol, glutamate, along with key genes related to trehalose, tryptophan and cysteine synthesis. Taken together, SAESo11-inoculation provides systemic effects encompassing both metabolic and regulatory functions, supporting both seedling growth and drought stress amelioration.


Subject(s)
Pseudomonas , Solanum lycopersicum , Pseudomonas/physiology , Transcriptome , Drought Resistance , Solanum lycopersicum/genetics , Metabolome , Droughts , Stress, Physiological/genetics
10.
Sci Rep ; 12(1): 13309, 2022 08 03.
Article in English | MEDLINE | ID: mdl-35922642

ABSTRACT

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.


Subject(s)
Bass , Fish Diseases , Pseudomonas Infections , Animals , Bacterial Proteins/metabolism , Bass/genetics , Fish Diseases/genetics , Pseudomonas/physiology , Pseudomonas Infections/genetics
11.
Invest Ophthalmol Vis Sci ; 63(2): 21, 2022 02 01.
Article in English | MEDLINE | ID: mdl-35142786

ABSTRACT

Purpose: To determine the possible microbiome related to Vogt-Koyanagi-Harada (VKH) disease in comparison to patients with noninfectious anterior scleritis and healthy people. Methods: Fecal samples were extracted from 42 individuals, including 11 patients with active VKH, 11 healthy people, and 20 patients with noninfectious anterior scleritis. We amplified the V3 to V4 16S ribosomal DNA (rDNA) region to obtain the target sequence. Then, the target sequence was amplified by polymerase chain reaction. The obtained target sequences were sequenced by high-throughput 16S rDNA analysis. Results: At the genus level, there were three enriched (Stomatobaculum, Pseudomonas, Lachnoanaerobaculum) and two depleted (Gordonibacter, Slackia) microbes that were detected only in patients with VKH. There were 10 enriched and 12 depleted microbes that were observed in both patients with VKH disease and noninfectious anterior scleritis (P < 0.05). The interactions of these microbes were graphed. Tyzzerella and Eggerthella were the nodes of interaction between these microorganisms, which were regulated by both positive and negative aspects, but the expression level in patients with active VKH was upregulated. Conclusions: Special or nonspecial enrichment and decreased intestinal microbes were observed in patients with active VKH. The action mechanism of these microbes needs further study.


Subject(s)
Actinobacteria/physiology , Clostridiales/physiology , Gastrointestinal Microbiome/physiology , Pseudomonas/physiology , Uveomeningoencephalitic Syndrome/microbiology , Adult , Case-Control Studies , DNA, Bacterial/genetics , Dysbiosis/microbiology , Feces/microbiology , Female , Genotyping Techniques , Healthy Volunteers , Humans , Male , Middle Aged , RNA, Ribosomal, 16S/genetics , Scleritis/microbiology
12.
Cells ; 10(12)2021 11 26.
Article in English | MEDLINE | ID: mdl-34943828

ABSTRACT

Drosophila suzukii is a neobiotic invasive pest that causes extensive damage to fruit crops worldwide. The biological control of this species has been unsuccessful thus far, in part because of its robust cellular innate immune system, including the activity of professional phagocytes known as hemocytes and plasmatocytes. The in vitro cultivation of primary hemocytes isolated from D. suzukii third-instar larvae is a valuable tool for the investigation of hemocyte-derived effector mechanisms against pathogens such as wasp parasitoid larvae, bacteria, fungi and viruses. Here, we describe the morphological characteristics of D. suzukii hemocytes and evaluate early innate immune responses, including extracellular traps released against the entomopathogen Pseudomonas entomophila and lipopolysaccharides. We show for the first time that D. suzukii plasmatocytes cast extracellular traps to combat P. entomophila, along with other cell-mediated reactions, such as phagocytosis and the formation of filopodia.


Subject(s)
Drosophila/immunology , Drosophila/microbiology , Extracellular Traps/metabolism , Immunity, Innate , Introduced Species , Pseudomonas/physiology , Animals , Cell Survival/drug effects , Drosophila/ultrastructure , Extracellular Traps/drug effects , Hemocytes/drug effects , Hemocytes/ultrastructure , Immunity, Innate/drug effects , Larva/cytology , Lipopolysaccharides/pharmacology , Phagocytes/drug effects , Phagocytes/microbiology , Pseudomonas/drug effects , Pseudopodia/drug effects , Pseudopodia/metabolism
13.
Int J Mol Sci ; 22(15)2021 Jul 23.
Article in English | MEDLINE | ID: mdl-34360661

ABSTRACT

Fabricated ecosystems (EcoFABs) offer an innovative approach to in situ examination of microbial establishment patterns around plant roots using nondestructive, high-resolution microscopy. Previously high-resolution imaging was challenging because the roots were not constrained to a fixed distance from the objective. Here, we describe a new 'Imaging EcoFAB' and the use of this device to image the entire root system of growing Brachypodium distachyon at high resolutions (20×, 40×) over a 3-week period. The device is capable of investigating root-microbe interactions of multimember communities. We examined nine strains of Pseudomonas simiae with different fluorescent constructs to B. distachyon and individual cells on root hairs were visible. Succession in the rhizosphere using two different strains of P. simiae was examined, where the second addition was shown to be able to establish in the root tissue. The device was suitable for imaging with different solid media at high magnification, allowing for the imaging of fungal establishment in the rhizosphere. Overall, the Imaging EcoFAB could improve our ability to investigate the spatiotemporal dynamics of the rhizosphere, including studies of fluorescently-tagged, multimember, synthetic communities.


Subject(s)
Brachypodium/microbiology , Microtechnology/instrumentation , Molecular Imaging/methods , Plant Roots/microbiology , Pseudomonas/physiology , Rhizosphere , Brachypodium/metabolism , Plant Roots/metabolism , Soil Microbiology
14.
Proc Natl Acad Sci U S A ; 118(35)2021 08 31.
Article in English | MEDLINE | ID: mdl-34446553

ABSTRACT

Polychlorinated biphenyls (PCBs) are persistent organic pollutants with severe effects on human health and the biosphere. Plant-based remediation offers many benefits over conventional PCB remediation, but its development has been hampered by our poor understanding of biphenyl metabolism in eukaryotes, among other factors. We report here a major PCB-responsive protein in poplar, a plant model system capable of PCB uptake and translocation. We provide structural and functional evidence that this uncharacterized protein, termed SDR57C, belongs to the heterogeneous short-chain dehydrogenase reductase (SDR) superfamily. Despite sequence divergence, structural modeling hinted at structural and functional similarities between SDR57C and BphB, a central component of the Bph pathway for biphenyl/PCB degradation in aerobic bacteria. By combining gas chromatography/mass spectrometry (GC/MS) profiling with a functional complementation scheme, we found that poplar SDR57C can replace BphB activity in the upper Bph pathway of Pseudomonas furukawaii KF707 and therefore catalyze the oxidation of 2,3-dihydro-2,3-dihydroxybiphenyl (2,3-DHDB) to 2,3-dihydroxybiphenyl (2,3-DHB). Consistent with this biochemical activity, we propose a mechanism of action based on prior quantum studies, general properties of SDR enzymes, and the modeled docking of 2,3-DHDB to the SDR57C-NAD+ complex. The putative detoxifying capacity of SDR57C was substantiated through reverse genetics in Arabidopsis thaliana Phenotypic characterization of the SDR lines underscored an inducible plant pathway with the potential to catabolize toxic biphenyl derivatives. Partial similarities with aerobic bacterial degradation notwithstanding, real-time messenger RNA quantification indicates the occurrence of plant-specific enzymes and features. Our results may help explain differences in degradative abilities among plant genotypes and also provide elements to improve them.


Subject(s)
Arabidopsis/drug effects , Biodegradation, Environmental , Plant Proteins/metabolism , Polychlorinated Biphenyls/metabolism , Populus/enzymology , Pseudomonas/physiology , Short Chain Dehydrogenase-Reductases/metabolism , Arabidopsis/growth & development , Arabidopsis/microbiology , Plant Proteins/genetics , Short Chain Dehydrogenase-Reductases/genetics
15.
Cell Rep ; 36(4): 109449, 2021 07 27.
Article in English | MEDLINE | ID: mdl-34320359

ABSTRACT

Bacterial communities are in a continuous adaptive and evolutionary race for survival. In this work we expand our knowledge on the chemical interplay and specific mutations that modulate the transition from antagonism to co-existence between two plant-beneficial bacteria, Pseudomonas chlororaphis PCL1606 and Bacillus amyloliquefaciens FZB42. We reveal that the bacteriostatic activity of bacillaene produced by Bacillus relies on an interaction with the protein elongation factor FusA of P. chlororaphis and how mutations in this protein lead to tolerance to bacillaene and other protein translation inhibitors. Additionally, we describe how the unspecific tolerance of B. amyloliquefaciens to antimicrobials associated with mutations in the glycerol kinase GlpK is provoked by a decrease of Bacillus cell membrane permeability, among other pleiotropic responses. We conclude that nutrient specialization and mutations in basic biological functions are bacterial adaptive dynamics that lead to the coexistence of two primary competitive bacterial species rather than their mutual eradication.


Subject(s)
Adaptation, Physiological , Bacillus/physiology , Pseudomonas/physiology , Adaptation, Physiological/drug effects , Alleles , Anti-Infective Agents/pharmacology , Bacillus/drug effects , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Drug Resistance, Bacterial/drug effects , Microbial Sensitivity Tests , Mutation/genetics , Permeability , Pseudomonas/drug effects , Pseudomonas/growth & development
16.
Nat Commun ; 12(1): 3829, 2021 06 22.
Article in English | MEDLINE | ID: mdl-34158504

ABSTRACT

While beneficial plant-microbe interactions are common in nature, direct evidence for the evolution of bacterial mutualism is scarce. Here we use experimental evolution to causally show that initially plant-antagonistic Pseudomonas protegens bacteria evolve into mutualists in the rhizosphere of Arabidopsis thaliana within six plant growth cycles (6 months). This evolutionary transition is accompanied with increased mutualist fitness via two mechanisms: (i) improved competitiveness for root exudates and (ii) enhanced tolerance to the plant-secreted antimicrobial scopoletin whose production is regulated by transcription factor MYB72. Crucially, these mutualistic adaptations are coupled with reduced phytotoxicity, enhanced transcription of MYB72 in roots, and a positive effect on plant growth. Genetically, mutualism is associated with diverse mutations in the GacS/GacA two-component regulator system, which confers high fitness benefits only in the presence of plants. Together, our results show that rhizosphere bacteria can rapidly evolve along the parasitism-mutualism continuum at an agriculturally relevant evolutionary timescale.


Subject(s)
Arabidopsis/genetics , Plant Roots/genetics , Pseudomonas/genetics , Rhizosphere , Symbiosis/genetics , Arabidopsis/growth & development , Arabidopsis/microbiology , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Evolution, Molecular , Gene Expression Regulation, Plant , Genetic Fitness , Host-Pathogen Interactions/genetics , Mutation , Phenotype , Plant Roots/growth & development , Plant Roots/microbiology , Pseudomonas/physiology , Transcription Factors/genetics , Transcription Factors/metabolism
17.
Mar Biotechnol (NY) ; 23(3): 463-471, 2021 Jun.
Article in English | MEDLINE | ID: mdl-34076776

ABSTRACT

Studies of invertebrates have shown that the internal environment of crustaceans is not always sterile in normal conditions, and in many species, it can be populated by microorganisms even in the absence of any visible pathological processes in the body. This observation raises the question of whether genetically modified indigenous hemolymph microorganisms can be used for biotechnological purposes inside the crustacean either as local producers of some compounds or as sensors to physiological parameters. In this study, we tested the ability of the bacteria isolated from the hemolymph of the amphipod Eulimnogammarus verrucosus to hide from the cellular immune response of the host as the most important feature for their potential long-term application in vivo. 16S rDNA amplicon sequencing revealed five common bacterial genera in all analyzed samples of the amphipod hemolymph, among which Pseudomonas is most easily subjected to genome modification and, thus, the most prospective for biotechnological application. Cultivation of Pseudomonas gave us a number of strains undoubtedly derived from the amphipod hemolymph, and one of them (belonging to the Pseudomonas fluorescens group) was chosen for further tests. The primary culture of amphipod hemocytes was used to analyze the immunogenicity of the strain and showed a pronounced reaction of the immune cells to a high amount of the bacteria within six hours. This result indicates that modulation of cellular immune response to metabolically active bacterial cells is not mandatory for the survival and wide distribution of these microorganisms in the hemolymph of numerous amphipod individuals.


Subject(s)
Amphipoda/immunology , Amphipoda/microbiology , Immunity, Cellular , Pseudomonas/physiology , Animals , Hemocytes , Hemolymph/cytology , Hemolymph/microbiology , Lakes , Siberia
18.
World J Microbiol Biotechnol ; 37(6): 94, 2021 May 08.
Article in English | MEDLINE | ID: mdl-33963474

ABSTRACT

The application of plant growth-promoting bacteria in agricultural systems is an efficient and environment-friendly strategy to improve crop yields and maintain soil quality. However, as different soils have diverse and specific ecological characteristics and may represent adverse abiotic conditions, in vivo application requires the careful selection of the desired beneficial microorganisms. In this study we report Ensifer adhaerens SZMC 25856 and Pseudomonas resinovorans SZMC 25875 isolates recovered from glyphosate-treated soil to possess yet undiscovered plant growth-enhancing potential. The strains were found to promote the growth of tomato seedlings significantly, to have the ability of synthesizing indole-3-acetic acid and siderophores, to tolerate pH in the range of 6.59-7.96, salinity up to 12.5 g L-1 NaCl and drought up to 125 g L-1 polyethylene glycol 6000, as well as to survive in the presence of various pesticides including glyphosate, diuron, chlorotoluron, carbendazim and thiabendazole, and heavy metals such as Al, Fe, Mn, Zn, Pb and Cu. The plant growth-promoting traits of the examined E. adhaerens and P. resinovorans isolates and their tolerance to numerous abiotic stress factors make them promising candidates for application in different agricultural environments, including soils polluted with glyphosate.


Subject(s)
Glycine/analogs & derivatives , Pseudomonas/isolation & purification , Rhizobiaceae/isolation & purification , Solanum lycopersicum/growth & development , Glycine/pharmacology , Hydrogen-Ion Concentration , Indoleacetic Acids/metabolism , Solanum lycopersicum/microbiology , Metals, Heavy/pharmacology , Pseudomonas/metabolism , Pseudomonas/physiology , RNA, Ribosomal, 16S/genetics , Rhizobiaceae/metabolism , Rhizobiaceae/physiology , Seeds/growth & development , Seeds/microbiology , Siderophores/metabolism , Soil Microbiology , Stress, Physiological , Glyphosate
19.
Nat Plants ; 7(6): 814-825, 2021 06.
Article in English | MEDLINE | ID: mdl-34031541

ABSTRACT

Plants grown in natural soil are colonized by phylogenetically structured communities of microbes known as the microbiota. Individual microbes can activate microbe-associated molecular pattern (MAMP)-triggered immunity (MTI), which limits pathogen proliferation but curtails plant growth, a phenomenon known as the growth-defence trade-off. Here, we report that, in monoassociations, 41% (62 out of 151) of taxonomically diverse root bacterial commensals suppress Arabidopsis thaliana root growth inhibition (RGI) triggered by immune-stimulating MAMPs or damage-associated molecular patterns. Amplicon sequencing of bacterial 16S rRNA genes reveals that immune activation alters the profile of synthetic communities (SynComs) comprising RGI-non-suppressive strains, whereas the presence of RGI-suppressive strains attenuates this effect. Root colonization by SynComs with different complexities and RGI-suppressive activities alters the expression of 174 core host genes, with functions related to root development and nutrient transport. Furthermore, RGI-suppressive SynComs specifically downregulate a subset of immune-related genes. Precolonization of plants with RGI-suppressive SynComs, or mutation of one commensal-downregulated transcription factor, MYB15, renders the plants more susceptible to opportunistic Pseudomonas pathogens. Our results suggest that RGI-non-suppressive and RGI-suppressive root commensals modulate host susceptibility to pathogens by either eliciting or dampening MTI responses, respectively. This interplay buffers the plant immune system against pathogen perturbation and defence-associated growth inhibition, ultimately leading to commensal-host homeostasis.


Subject(s)
Arabidopsis/immunology , Host-Pathogen Interactions/physiology , Microbiota , Plant Immunity/physiology , Plant Roots/microbiology , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Arabidopsis Proteins/immunology , Gene Expression Regulation, Plant , Host-Pathogen Interactions/immunology , Pathogen-Associated Molecular Pattern Molecules , Phylogeny , Plant Diseases/immunology , Plant Diseases/microbiology , Plant Roots/genetics , Plant Roots/growth & development , Plants, Genetically Modified , Pseudomonas/physiology
20.
PLoS One ; 16(4): e0250574, 2021.
Article in English | MEDLINE | ID: mdl-33901241

ABSTRACT

In the present study we examined the effect of nanogypsum and Pseudomonas taiwanensis strain BCRC 17751on plant and soil health using conventional and metagenomics approaches. Soil physicochemical properties and agronomical parameters of maize plants were reported to be better when applied with nanogypsum and bacterial inoculum together. When compared to control a significant increase in total bacterial counts, nitrogen, phosphorus, potassium (NPK) solubilizing bacterial population and soil enzyme activities (fluorescein diacetate, alkaline phosphatase, dehydrogenase, ß-glucosidase, arylesterase and amylase) was reported in treatments. The metagenomics studies revealed dominance of beneficial bacteria such as Proteobacteria, Bacteriodetes, Planctomycetes, Acidobacteria and Nitrospirae in treated soil. On the other hand some novel bacterial diversity was also reported in treated soil which was evident from presence of taxonomically unclassified sequences. Hence, it can be concluded that combined application of nanogypsum and Pseudomonas taiwanensis in maize help in improving the structure and function of soil which affects the plant health without causing any toxic effect. However, in situ validation of the prescribed treatment is required under field conditions on different crops in order to give maximum benefits to the farmers and the environment.


Subject(s)
Calcium Sulfate/pharmacology , Metagenomics , Microbiota , Nanoparticles/chemistry , Pseudomonas/physiology , Rhizosphere , Zea mays/growth & development , Zea mays/microbiology , Agriculture , Crops, Agricultural/growth & development , Microbiota/drug effects , Nitrogen/analysis , Phosphorus/analysis , Phylogeny , Potassium/analysis , Pseudomonas/drug effects , Soil/chemistry , Zea mays/drug effects
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