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1.
J Water Health ; 22(6): 1033-1043, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38935454

ABSTRACT

The misuse of antibiotics and the emergence of antimicrobial resistance (AMR) is a concern in the aquaculture industry because it contributes to global health risks and impacts the environment. This study analyzed the AMR of sentinel bacteria associated with striped catfish (Pangasisanodon hypophthalmus) and giant snakehead (Channa micropeltes), the two main fish species reared in the pond culture in Cambodia. Phenotypic and genotypic characterization of the recovered isolates from fish, water, and sediment samples revealed the presence of bacteria, such as 22 species belonging to families Aeromonadaceae, Enterobacteriaceae, and Pseudomonadaceae. Among 48 isolates, Aeromonas caviae (n = 2), Aeromonas hydrophila (n = 2), Aeromonas ichthiosmia (n = 1), Aeromonas salmonicida (n = 4) were detected. A. salmonicida and A. hydrophilla are known as fish pathogens that occur worldwide in both fresh and marine water aquaculture. Antibiotic susceptibility testing revealed antibiotic resistance patterns of 24 (50 %) isolates among 48 isolates with higher multiple antibiotic resistance index (> 0.2). All the isolates of Enterobacteriaceae were susceptible to ciprofloxacin. Ciprofloxacin is a frontline antibiotic that is not recommended to use in aquaculture. Therefore, its use has to be strictly controlled. This study expands our knowledge of the AMR status in aquaculture farms which is very limited in Cambodia.


Subject(s)
Aquaculture , Drug Resistance, Bacterial , Water Microbiology , Cambodia , Catfishes/microbiology , Sentinel Species , Phenotype , Genotype , Aeromonadaceae/classification , Aeromonadaceae/isolation & purification , Aeromonadaceae/physiology , Enterobacteriaceae/classification , Enterobacteriaceae/isolation & purification , Enterobacteriaceae/physiology , Pseudomonadaceae/classification , Pseudomonadaceae/isolation & purification , Pseudomonadaceae/physiology , Aeromonas caviae/isolation & purification , Aeromonas caviae/physiology , Aeromonas hydrophila/isolation & purification , Aeromonas hydrophila/physiology , Ciprofloxacin/pharmacology , Ciprofloxacin/therapeutic use , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Fish Diseases/drug therapy , Fish Diseases/microbiology , Gram-Negative Bacterial Infections/drug therapy , Gram-Negative Bacterial Infections/veterinary , Environmental Monitoring
2.
Microbiol Spectr ; 11(4): e0115023, 2023 08 17.
Article in English | MEDLINE | ID: mdl-37404179

ABSTRACT

Ginseng is a popular medicinal herb with established therapeutic effects such as cardiovascular disease prevention, anticancer effects, and anti-inflammatory effects. However, the slow growth of ginseng due to soilborne pathogens has been a challenge for establishing new plantations. In this study, we investigated root rot disease associated with the microbiota in a ginseng monoculture model system. Our results showed that a collapse of the early microbiota community inhibiting root rot disease was observed before the disease became severe, and nitrogen fixation was necessary to support the initial microbiota community structure. Furthermore, changes in the nitrogen composition were essential for the suppression of pathogen activity in early monoculture soils. We hypothesize that Pseudomonadaceae, a population built up by aspartic acid, can inhibit the occurrence of root rot disease in ginseng and that specific management practices that maintain a healthy microbiome can be implemented to prevent and mitigate the disease. Our findings provide insights into the potential use of specific members of the microbiota for controlling root rot disease in ginseng cultivation. IMPORTANCE Understanding the initial soil microbiota and community shifts in a monoculture system is critical for developing disease-suppressive soils for crop production. The lack of resistance genes against soilborne pathogens in plants highlights the need for effective management strategies. Our investigation of root rot disease and initial microbiota community shifts in a ginseng monoculture model system provides valuable insight into the development of conducive soil into specific suppressive soil. With a thorough understanding of the microbiota in disease-conducive soil, we can work toward the development of disease-suppressive soil to prevent outbreaks and ensure sustainable crop production.


Subject(s)
Panax , Soil , Soil/chemistry , Pseudomonadaceae , Soil Microbiology , Plant Diseases/prevention & control
3.
Article in English | MEDLINE | ID: mdl-36862578

ABSTRACT

Two bacterial strains, designated 5GH9-11T and 5GH9-34T, were isolated from greenhouse soil sampled in Wanju-gun, Jeollabuk-do, Republic of Korea. Both strains formed yellow colonies and were aerobic, rod-shaped and flagellated. The 16S rRNA gene sequence similarity between 5GH9-11T and 5GH9-34T was 98.6 %. Strain 5GH9-11T showed the highest sequence similarities to Dyella thiooxydans ATSB10T (98.1 %) and Frateuria aurantia DSM 6220T (97.7 %) while strain 5GH9-34T revealed the highest sequence similarity to F. aurantia DSM 6220T (98.3 %) and D. thiooxydans ATSB10T (98.3 %). Phylogenetic analysis on the basis of the 16S rRNA gene sequence showed that strains 5GH9-11T and 5GH9-34T formed a robust cluster with Frateuria flava MAH-13T and Frateuria terrea NBRC 104236T. The phylogenomic tree also showed that strains 5GH9-11T and 5GH9-34T formed a robust cluster with F. terrea DSM 26515T and F. flava MAH-13T. Strain 5GH9-11T showed the highest orthologous average nucleotide identity (OrthoANI; 88.5 %) and digital DNA-DNA hybridization (dDDH) values (35.5 %) with F. flava MAH-13T, and strain 5GH9-34T revealed highest OrthoANI (88.1 %) and dDDH (34.2 %) values with F. flava MAH-13T. The orthoANI and dDDH values between strain 5GH9-11T and 5GH9-34T were 87.7 and 33.9 %, respectively. Their major respiratory quinone was ubiquinone 8, and the major cellular fatty acids were iso-C16 : 0, summed feature 9 (iso-C17 : 1 ω9c and/or C16 : 0 10-methyl) and iso-C15 : 0. The major polar lipids of both strains were composed of large or moderate amounts of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified aminolipid and an unidentified aminophospholipid. Based on these data, strains 5GH9-11T and 5GH9-34T should represent two independent novel species of Frateuria, for which the names Frateuria soli sp. nov. (type strain 5GH9-11T=KACC 16943T=JCM 35197T) and Frateuria edaphi sp. nov. (type strain 5GH9-34T=KACC 16945T=JCM 35198T) are proposed.


Subject(s)
Fatty Acids , Pseudomonadaceae , Phylogeny , RNA, Ribosomal, 16S/genetics , Fatty Acids/chemistry , Sequence Analysis, DNA , DNA, Bacterial/genetics , Bacterial Typing Techniques , Base Composition
5.
Viruses ; 14(2)2022 01 26.
Article in English | MEDLINE | ID: mdl-35215838

ABSTRACT

The phyllosphere microbiome plays an important role in plant fitness. Recently, bacteriophages have been shown to play a role in shaping the bacterial community composition of the phyllosphere. However, no studies on the diversity and abundance of phyllosphere bacteriophage communities have been carried out until now. In this study, we extracted, sequenced, and characterized the dsDNA and ssDNA viral community from a phyllosphere for the first time. We sampled leaves from winter wheat (Triticum aestivum), where we identified a total of 876 virus operational taxonomic units (vOTUs), mostly predicted to be bacteriophages with a lytic lifestyle. Remarkably, 848 of these vOTUs corresponded to new viral species, and we estimated a minimum of 2.0 × 106 viral particles per leaf. These results suggest that the wheat phyllosphere harbors a large and active community of novel bacterial viruses. Phylloviruses have potential applications as biocontrol agents against phytopathogenic bacteria or as microbiome modulators to increase plant growth-promoting bacteria.


Subject(s)
Bacteriophages/isolation & purification , Triticum/microbiology , Bacteriophages/classification , Bacteriophages/genetics , Genome, Viral/genetics , Metagenome/genetics , Microbiota , Plant Leaves/microbiology , Pseudomonadaceae/classification , Pseudomonadaceae/genetics , Pseudomonadaceae/isolation & purification , Pseudomonadaceae/virology , Toxins, Biological/genetics
6.
Chem Biodivers ; 18(12): e2100516, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34609783

ABSTRACT

Bacterial endophytes are known to protect Vitis vinifera L. against various harmful effects of the environment and support its growth. However, for the most part, biochemical responses of such co-existence have not yet been fully elucidated. In this work, we aimed to characterize the activities of endophytic consortia in a plant-endophyte extract by measuring five indicators of colonization (overall endophyte metabolic activity, microbial ACC deaminase activity, ability to solubilize phosphorus, ability to convert atmospheric nitrogen to ammonia ions, and ability to produce growth promoting indole acetic acid), and find relationships between these activities and metabolome. The V. vinifera canes for the metabolomics fingerprinting were extracted successively with water and methanol, and analysed by ultra-high performance liquid chromatography coupled with high resolution mass spectrometry. For data processing, the MS-DIAL - MS-CleanR - MS-FINDER software platform was used, and the data matrix was processed by PCA and PLS-DA multivariate statistical methods. The metabolites that were upregulated with the heavy endophyte colonization were mainly chlorins, phenolics, flavonoid and terpenoid glycosides, tannins, dihydropyranones, sesquiterpene lactones, and long-chain unsaturated fatty acids.


Subject(s)
Endophytes/metabolism , Metabolomics , Vitis/chemistry , Bacillaceae/metabolism , Enterobacteriaceae/metabolism , Micrococcaceae/metabolism , Pseudomonadaceae/metabolism , Vitis/metabolism
7.
Article in English | MEDLINE | ID: mdl-34546867

ABSTRACT

The evolutionary relationships among species of the family Pseudomonadaceae were examined based on 255 available genomes representing >85 % of the species from this family. In a phylogenetic tree based on concatenated sequences of 118 core proteins, most species of the genus Pseudomonas grouped within one large cluster which also included members of the genera Azotobacter and Azomonas. Within this large cluster 18-30 clades/subclades of species of the genus Pseudomonas consisting of between 1 and 36 species, were observed. However, a number of species of the genus Pseudomonas branched outside of this main cluster and were interspersed among other genera of the family Pseudomonadaceae. This included a strongly supported clade (Pertucinogena clade) consisting of 19 mainly halotolerant species. The distinctness of this clade from all other members of the family Pseudomonadaceae is strongly supported by 24 conserved signature indels (CSIs) in diverse proteins that are exclusively found in all members of this clade. Nine uncharacterized members of the genus Pseudomonas also shared these CSIs and they branched within the Pertucinogena clade, indicating their affiliation to this clade. On the basis of the strong evidence supporting the distinctness of the Pertucinogena clade, we are proposing transfer of species from this clade into a novel genus Halopseudomonas gen. nov. Pseudomonas caeni also branches outside of the main cluster and groups reliably with Oblitimonas alkaliphila and Thiopseudomonas denitrificans. Six identified CSIs are uniquely shared by these three species and we are proposing their integration into the emended genus Thiopseudomonas, which has priority over the name Oblitimonas. We are also proposing transfer of the deep-branching Pseudomonas hussainii, for which 22 exclusive CSIs have been identified, into the genus Atopomonas gen. nov. Lastly, we present strong evidence that the species Pseudomonas cissicola and Pseudomonas geniculata are misclassified into the genus Pseudomonas and that they are specifically related to the genera Xanthomonas and Stenotrophomonas, respectively. In addition, we are also reclassifying 'Pseudomonas acidophila' as Paraburkholderia acidicola sp. nov. (Type strain: G-6302=ATCC 31363=BCRC 13035).


Subject(s)
Fatty Acids , Genomics , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/chemistry , Phylogeny , Pseudomonadaceae , Pseudomonas/genetics , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Xanthomonas
8.
Article in English | MEDLINE | ID: mdl-34516367

ABSTRACT

Strain F2AT, isolated from the cricket Acheta domesticus, was subjected to a polyphasic taxonomic characterization. Cells of the strain were rod-shaped, Gram-stain-negative and catalase- and oxidase-positive. It did not assimilate any carbohydrates. The strain's 16S rRNA gene sequence showed highest similarity to Entomomonas moraniae QZS01T (96.4 %). The next highest similarity values were found to representatives of related genera (<93 %). The genome size of strain F2AT was 3.2 Mbp and the G+C content was 36.4 mol%. Average nucleotide identity values based on blast and MUMmer and average amino acid identity values between strain F2AT and E. moraniae QZS01T were 74.29/74.43, 83.88 and 74.70 %, respectively. The quinone system predominantly contained ubiquinone Q-8. In the polar lipid profile, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and an unidentified phospholipid were detected. The polyamine pattern consisted of the major compounds putrescine and spermidine. Major fatty acids were C18 : 1 ω7c and C16 : 0 and the hydroxyl acids were C12 : 0 3-OH, C14 : 0 2-OH and C14 : 0 3-OH. The diagnostic diamino acid of the peptidoglycan was meso-diaminopimelic acid. Due to its association with the only species of the genus Entomomonas but its distinctness from E. moraniae we here propose the novel species Entomomonas asaccharolytica sp. nov. F2AT (=CCM 9136T=LMG 32211T).


Subject(s)
Gryllidae , Phylogeny , Pseudomonadaceae/classification , Animals , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Diaminopimelic Acid/chemistry , Fatty Acids/chemistry , Gryllidae/microbiology , Nucleic Acid Hybridization , Peptidoglycan/chemistry , Phospholipids/chemistry , Pseudomonadaceae/isolation & purification , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Ubiquinone/chemistry
9.
Microbiol Spectr ; 9(1): e0045221, 2021 09 03.
Article in English | MEDLINE | ID: mdl-34378950

ABSTRACT

A group of 11 bacterial strains was isolated from streams and lakes located in a deglaciated northern part of James Ross Island, Antarctica. They were rod-shaped, Gram-stain-negative, motile, and catalase-positive and produced blue-violet-pigmented colonies on R2A agar. A polyphasic taxonomic approach based on 16S rRNA gene sequencing, whole-genome sequencing, automated ribotyping, repetitive element sequence-based PCR (rep-PCR), MALDI-TOF MS, fatty acid profile, chemotaxonomy analyses, and extensive biotyping was applied in order to clarify the taxonomic position of these isolates. Phylogenetic analysis based on the 16S rRNA gene indicated that all the isolates constituted a coherent group belonging to the genus Rugamonas. The closest relatives to the representative isolate P5900T were Rugamonas rubra CCM 3730T, Rugamonas rivuli FT103WT, and Rugamonas aquatica FT29WT, exhibiting 99.2%, 99.1%, and 98.6% 16S rRNA pairwise similarity, respectively. The average nucleotide identity and digital DNA-DNA hybridization values calculated from the whole-genome sequencing data clearly proved that P5900T represents a distinct Rugamonas species. The G+C content of genomic DNAs was 66.1 mol%. The major components in fatty acid profiles were summed feature 3 (C16:1ω7c/C16:1ω6c), C 16:0, and C12:0. The cellular quinone content contained exclusively ubiquinone Q-8. The predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine. The polyamine pattern was composed of putrescine, 2-hydroxputrescine, and spermidine. IMPORTANCE Our polyphasic approach provides a new understanding of the taxonomy of novel pigmented Rugamonas species isolated from freshwater samples in Antarctica. The isolates showed considerable extracellular bactericidal secretions. The antagonistic activity of studied isolates against selected pathogens was proved by this study and implied the importance of such compounds' production among aquatic bacteria. The psychrophilic and violacein-producing species Roseomonas violacea may play a role in the diverse consortium among pigmented bacteria in the Antarctic water environment. Based on all the obtained results, we propose a novel species for which the name Rugamonas violacea sp. nov. is suggested, with the type strain P5900T (CCM 8940T; LMG 32105T). Isolates of R. violacea were obtained from different aquatic localities, and they represent the autochthonous part of the water microbiome in Antarctica.


Subject(s)
Indoles/metabolism , Phylogeny , Pseudomonadaceae/classification , Pseudomonadaceae/isolation & purification , Pseudomonadaceae/metabolism , Antarctic Regions , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Lakes , Pseudomonadaceae/genetics , RNA, Ribosomal, 16S/genetics , Soil Microbiology
10.
Nat Plants ; 7(5): 644-654, 2021 05.
Article in English | MEDLINE | ID: mdl-33972713

ABSTRACT

Maintaining microbiome structure is critical for the health of both plants and animals. By re-screening a collection of Arabidopsis mutants affecting root immunity and hormone crosstalk, we identified a FERONIA (FER) receptor kinase mutant (fer-8) with a rhizosphere microbiome enriched in Pseudomonas fluorescens without phylum-level dysbiosis. Using microbiome transplant experiments, we found that the fer-8 microbiome was beneficial. The effect of FER on rhizosphere pseudomonads was largely independent of its immune scaffold function, role in development and jasmonic acid autoimmunity. We found that the fer-8 mutant has reduced basal levels of reactive oxygen species (ROS) in roots and that mutants deficient in NADPH oxidase showed elevated rhizosphere pseudomonads. The addition of RALF23 peptides, a FER ligand, was sufficient to enrich P. fluorescens. This work shows that FER-mediated ROS production regulates levels of beneficial pseudomonads in the rhizosphere microbiome.


Subject(s)
Arabidopsis Proteins/physiology , Phosphotransferases/physiology , Pseudomonas fluorescens/metabolism , Reactive Oxygen Species/metabolism , Rhizosphere , Soil Microbiology , Arabidopsis/metabolism , Arabidopsis Proteins/metabolism , Phosphotransferases/metabolism , Pseudomonadaceae/metabolism , Pseudomonadaceae/physiology , Pseudomonas fluorescens/physiology
11.
Article in English | MEDLINE | ID: mdl-33881985

ABSTRACT

A Gram-stain-negative, aerobic, non-motile and rod-shaped bacterium, designated as IMCC34836T, was isolated from a freshwater stream. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain IMCC34836T was most closely related to Permianibacter aggregans HW001T (of the family Pseudomonadaceae) with 95.6 % sequence similarity and formed a robust clade with P. aggregans HW001T. The draft genome sequence of strain IMCC34836T was 4.4 Mbp in size with 59.1 mol% DNA G+C content. Average nucleotide identity and digital DNA-DNA hybridization values between strain IMCC34836T and P. aggregans HW001T were 71.2 and 22.0 %, respectively, indicating that the new strain represents a novel species. The strain contained iso-C15 : 0, summed feature 3 (C16 : 1 ω6c and/or C16 : 1 ω7c) and summed feature 9 (iso-C17 : 1 ω9c and/or C16 : 1 10-methyl) as the major fatty acids and harboured phosphatidylethanolamine, two unidentified aminophospholipids and three unidentified lipids as major polar lipids. The isoprenoid quinone detected in the strain was ubiquinone-8. Based on the phylogenetic and phenotypic characteristics, strain IMCC34836T is considered to represent a novel species of the genus Permianibacter, for which the name Permianibacter fluminis sp. nov. is proposed. The type strain is IMCC34836T (=KACC 21755T=NBRC 114416T).


Subject(s)
Phylogeny , Pseudomonadaceae/classification , Rivers/microbiology , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/chemistry , Nucleic Acid Hybridization , Phosphatidylethanolamines , Phospholipids/chemistry , Pseudomonadaceae/isolation & purification , RNA, Ribosomal, 16S/genetics , Republic of Korea , Sequence Analysis, DNA , Ubiquinone/chemistry
12.
Anim Sci J ; 91(1): e13441, 2020.
Article in English | MEDLINE | ID: mdl-32885570

ABSTRACT

Microbiota of individual cow milk, bulk tank milk, and feces of Jersey cows were examined. Samples were collected from two farms (F1 and F2) in cool (November, Nov) and hot (July, Jul) seasons. Milk yield and milk composition were similar between the two farms and between the two seasons. Prevalent taxa of the fecal microbiota, i.e. Ruminococcaceae, Bacteroidaceae, Lachnospiraceae, Rikenellaceae, and Clostridiaceae, were unaffected by the farm and season. Relative abundance of milk microbiota for Pseudomonadaceae, Enterobacteriaceae, and Streptococcaceae (F1 > F2) and Lactobacillaceae, Bifidobacteriaceae, and Cellulomonadaceae (F1 < F2) were different between the two farms, and those for Staphylococcaceae, Bacillaceae, Ruminococcaceae, and Veillonellaceae (Nov < Jul) and Methylobacteriaceae and Moraxellaceae (Nov > Jul) were different between the two seasons. The microbiota of bulk tank milk was numerically different from that of individual cow milk. Principal coordinate analysis indicated that the milk microbiota was unrelated to the fecal microbiota. The finding that relative abundance of Pseudomonadaceae and Moraxellaceae appeared greater than those reported for Holstein milk suggested that higher protein and fat content may result in a greater abundance of proteolytic and lipolytic taxa in Jersey cow milk.


Subject(s)
Cattle/metabolism , Cattle/microbiology , Cold Temperature , Dairying , Farms , Fats/metabolism , Feces/microbiology , Hot Temperature , Milk Proteins/metabolism , Milk/metabolism , Milk/microbiology , Seasons , Animals , Female , Moraxellaceae , Proteolysis , Pseudomonadaceae
13.
J Microbiol Methods ; 172: 105911, 2020 05.
Article in English | MEDLINE | ID: mdl-32240707

ABSTRACT

Pseudomonas protegens is a rhizosphere pseudomonad with a high agronomical potential (entomopathogenic and beneficial to plants) and bio-catalytic activities, but no selective medium has been described for its isolation. We developed a semi-selective minimum agar medium for the specific isolation and growth of P. protegens. We searched for both (i) a carbon source allowing the growth of P. protegens but potentially inhibiting the growth of other pseudomonads and (ii) an antimicrobial agent suppressing other members of the bacterial rhizosphere community. The M9-PP-agar medium consists of M9 base agar with adipic acid as the only carbon source and Irgasan® as an anti-bacterial agent. We tested the selectivity and sensitivity of M9-PP-agar by measuring the growth of 68 bacterial strains from 36 different species on this medium. Ten of the species tested were able to grow on M9-PP-agar medium: four species from the Pseudomonadaceae (Pseudomonas aeruginosa, Pseudomonas protegens, Pseudomonas putida, Stenotrophomonas maltophilia) as well as Achromobacter xylosoxidans, Agrobacterium tumefaciens, Brevundimonas sp., Serratia liquefaciens, Serratia marcescens and Variovorax paradoxus. All colonies were white, except for those of P. protegens (12 strains), which were typically brown. We demonstrated the efficiency of the M9-PP agar medium for P. protegens isolation, by inoculating two soils with the reference strain P. protegens CHAOT and then reisolating them. We also developed a fitF-PCR test targeting a regulator gene of the insecticidal P. protegens fit locus, for the rapid molecular detection of P. protegens colonies. We, therefore, developed a highly specific process for the routine isolation of new P. protegens strains from the soil environment, based on the use of a semi-selective medium and the specific color of colonies.


Subject(s)
Bacteriological Techniques/methods , Culture Media/chemistry , Pseudomonas/isolation & purification , Soil Microbiology , Anti-Infective Agents/pharmacology , Bacteria/classification , Bacteria/isolation & purification , Carbon/metabolism , DNA, Bacterial/analysis , Gram-Negative Bacteria , Microbial Sensitivity Tests , Molecular Typing/methods , Polymerase Chain Reaction/methods , Pseudomonadaceae/classification , Pseudomonadaceae/isolation & purification , Pseudomonas/classification , Pseudomonas/drug effects , Rhizosphere , Soil
14.
Int J Syst Evol Microbiol ; 70(4): 2426-2434, 2020 Apr.
Article in English | MEDLINE | ID: mdl-32068524

ABSTRACT

As the current episode of Acute Oak Decline (AOD) continues to affect native British oak in the United Kingdom, ongoing isolations from symptomatic and healthy oak have yielded a large Pseudomonas species population. These strains could be divided into taxa representing three potential novel species. Recently, two of these taxa were described as novel Pseudomonas species in the Pseudomonas fluorescens lineage. Here, we demonstrate using a polyphasic approach that the third taxon represents another novel Pseudomonas species. The 16S rRNA gene sequencing assigned the strains to the Pseudomonas aeruginosa lineage, while multilocus sequence analysis (based on partial gyrB, rpoB and rpoD sequences) placed the 13 strains in a single cluster on the border of the Pseudomonas stutzeri group. Whole genome intra-species comparisons (based on average nucleotide identity and in silico DNA-DNA hybridization) confirmed that the strains belong to a single taxon, while the inter-species comparisons with closest phylogenetic relatives yielded similarity values below the accepted species threshold. Therefore, we propose these strains as a novel species, namely Pseudomonas kirkiae sp. nov., with the type strain FRB 229T (P4CT=LMG 31089T=NCPPB 4674T). The phylogenetic analyses performed in this study highlighted the difficulties in assigning novel species to the genus Pseudomonas due to its polyphyletic nature and close relationship to the genus Azotobacter. We further propose that a thorough taxonomic re-evaluation of the genus Pseudomonas is essential and should be performed in the near future.


Subject(s)
Azotobacter/classification , Phylogeny , Plant Diseases/microbiology , Pseudomonadaceae/classification , Pseudomonas/classification , Quercus/microbiology , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Genes, Bacterial , Multilocus Sequence Typing , Nucleic Acid Hybridization , Pseudomonas/isolation & purification , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , United Kingdom
15.
Environ Microbiol ; 22(4): 1447-1466, 2020 04.
Article in English | MEDLINE | ID: mdl-32011068

ABSTRACT

Siderophores are iron-chelating molecules produced by bacteria to access iron, a key nutrient. These compounds have highly diverse chemical structures, with various chelating groups. They are released by bacteria into their environment to scavenge iron and bring it back into the cells. The biosynthesis of siderophores requires complex enzymatic processes and expression of the enzymes involved is very finely regulated by iron availability and diverse transcriptional regulators. Recent data have also highlighted the organization of the enzymes involved in siderophore biosynthesis into siderosomes, multi-enzymatic complexes involved in siderophore synthesis. An understanding of siderophore biosynthesis is of great importance, as these compounds have many potential biotechnological applications because of their metal-chelating properties and their key role in bacterial growth and virulence. This review focuses on the biosynthesis of siderophores produced by fluorescent Pseudomonads, bacteria capable of colonizing a large variety of ecological niches. They are characterized by the production of chromopeptide siderophores, called pyoverdines, which give the typical green colour characteristic of fluorescent pseudomonad cultures. Secondary siderophores are also produced by these strains and can have highly diverse structures (such as pyochelins, pseudomonine, yersiniabactin, corrugatin, achromobactin and quinolobactin).


Subject(s)
Pseudomonadaceae/metabolism , Siderophores/biosynthesis , Iron/metabolism , Secondary Metabolism
16.
Int J Syst Evol Microbiol ; 70(1): 165-171, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31560340

ABSTRACT

The honey bee gut microbiota contains many bacterial lineages that are specific to this ecosystem. Apis cerana, raised across the Asian continent, is of great significance to the maintenance and development of ecology and agriculture in Asia. Here, we report the isolation and characterization of strain QZS01T from the gut of Apis cerana from Pingwu County, Sichuan Province, PR China. The results of phylogenetic analysis based on 16S rRNA sequences showed that strain QZS01T forms a monophyletic group together with clone sequences derived from variable insect hosts, and it shows 92% sequence similarity to its closest relative, Pseudomonas knackmussii. Strain QZS01T possesses a reduced genome (3.3 Mbp; G+C content, 38.05 mol%) compared to all other Pseudomonas species, and the whole-genome based phylogenetic reconstruction showed that strain QZS01T represents a novel genus within the family Pseudomonadaceae. Strain QZS01T is a Gram-stain-negative facultative anaerobe. It grows on brain heart infusion agar and the energy sources utilized for growth are very limited. Based on the results of genotypic and phenotypic analyses, we propose a novel genus and species, Entomomonas moraniae gen. nov., sp. nov., with the type strain QZS01T (=CGMCC 1.13498T=KCTC 62495T).


Subject(s)
Bees/microbiology , Gastrointestinal Tract/microbiology , Genome, Bacterial , Phylogeny , Pseudomonadaceae/classification , Animals , Bacterial Typing Techniques , Base Composition , China , DNA, Bacterial/genetics , Fatty Acids/chemistry , Pseudomonadaceae/isolation & purification , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
17.
Nat Prod Rep ; 37(1): 29-54, 2020 01 01.
Article in English | MEDLINE | ID: mdl-31436775

ABSTRACT

Bacteria of the genus Pseudomonas are ubiquitous in nature. Pseudomonads display a fascinating metabolic diversity, which correlates with their ability to colonize an extremely wide range of ecological niches. As a result, these bacteria are a prolific source of natural products. Biosynthesis of the latter is often orchestrated by arrays of chemical signals arising from intraspecies communication or interspecies relationships with bacteria, fungi, amoebae, plants, and insects. Especially nonribosomal lipopeptides, which have diverse biological activities, play important roles in the lifestyle of pseudomonads. In this review, we will focus on the molecular structures, properties, biosynthetic pathways, and biological functions of pseudomonal lipopeptides. This review is not only addressed to bio/chemists rather it serves as a comprehensive guide for all researchers (micro/biologists, ecologists, and environmental scientists) working in this multidisciplinary field.


Subject(s)
Lipopeptides/biosynthesis , Lipopeptides/chemistry , Lipopeptides/pharmacology , Pseudomonadaceae/metabolism , Host-Pathogen Interactions , Molecular Structure , Plants/microbiology , Pseudomonadaceae/physiology , Pseudomonas/metabolism , Pseudomonas/pathogenicity , Rhizosphere , Ribosomes/metabolism , Soil Microbiology
18.
Cell Mol Life Sci ; 77(3): 395-413, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31768608

ABSTRACT

Microorganisms acquire energy and nutrients from dynamic environments, where substrates vary in both type and abundance. The regulatory system responsible for prioritizing preferred substrates is known as carbon catabolite repression (CCR). Two broad classes of CCR have been documented in the literature. The best described CCR strategy, referred to here as classic CCR (cCCR), has been experimentally and theoretically studied using model organisms such as Escherichia coli. cCCR phenotypes are often used to generalize universal strategies for fitness, sometimes incorrectly. For instance, extremely competitive microorganisms, such as Pseudomonads, which arguably have broader global distributions than E. coli, have achieved their success using metabolic strategies that are nearly opposite of cCCR. These organisms utilize a CCR strategy termed 'reverse CCR' (rCCR), because the order of preferred substrates is nearly reverse that of cCCR. rCCR phenotypes prefer organic acids over glucose, may or may not select preferred substrates to optimize growth rates, and do not allocate intracellular resources in a manner that produces an overflow metabolism. cCCR and rCCR have traditionally been interpreted from the perspective of monocultures, even though most microorganisms live in consortia. Here, we review the basic tenets of the two CCR strategies and consider these phenotypes from the perspective of resource acquisition in consortia, a scenario that surely influenced the evolution of cCCR and rCCR. For instance, cCCR and rCCR metabolism are near mirror images of each other; when considered from a consortium basis, the complementary properties of the two strategies can mitigate direct competition for energy and nutrients and instead establish cooperative division of labor.


Subject(s)
Carbon/metabolism , Catabolite Repression/physiology , Pseudomonadaceae/metabolism , Escherichia coli/metabolism , Glucose/metabolism , Humans , Phenotype
19.
Biochim Biophys Acta Biomembr ; 1861(12): 183060, 2019 12 01.
Article in English | MEDLINE | ID: mdl-31499020

ABSTRACT

In recent years, hopanoids, a group of pentacyclic compounds found in bacterial membranes, are in the spotlight since it was proposed that they induce order in lipid membranes in a similar way cholesterol do in eukaryotes, despite their structural differences. We studied here whether diplopterol (an abundant hopanoid) promoted similar effects on model membranes as sterols do. We analyzed the compaction, dynamics, phase segregation, permeability and compressibility of model membranes containing diplopterol, and compared with those containing sterols from animals, plants and fungi. We also tested the effect that the incubation with diplopterol had on hopanoid-lacking bacteria. Our results show that diplopterol induces phase segregation, increases lipid compaction, and decreases permeability on phospholipid membranes, while retaining membrane fluidity and compressibility. Furthermore, the exposition to this hopanoid decreases the permeability of the opportunistic pathogen Pseudomonas aeruginosa and increases the resistance to antibiotics. All effects promoted by diplopterol were similar to those generated by the sterols. Our observations add information on the functional significance of hopanoids as molecules that play an important role in membrane organization and dynamics in model membranes and in a bacterial system.


Subject(s)
Cell Membrane Permeability/physiology , Cell Membrane/chemistry , Triterpenes/metabolism , Cell Membrane/physiology , Cell Membrane Permeability/drug effects , Lipid Bilayers/chemistry , Membrane Lipids/chemistry , Membrane Lipids/physiology , Membranes/chemistry , Membranes/physiology , Models, Biological , Permeability , Phospholipids/chemistry , Phospholipids/physiology , Pseudomonadaceae/metabolism , Sterols/chemistry , Triterpenes/pharmacology
20.
Mol Biol Rep ; 46(3): 3357-3370, 2019 Jun.
Article in English | MEDLINE | ID: mdl-30997598

ABSTRACT

About 24 h incubation of Azomonas (A.) macrocytogenes isolate KC685000 in 14L fermenter produced 22% poly (3-hydroxybutyrate) (PHB) per cell dry weight (CDW) biopolymer using 1 vvm aeration, 10% inoculum size, and initial pH of 7.2. To control the fermentation process, Logistic and Leudeking-Piret models were used to describe the cell growth and PHB production, respectively. These two models were in good agreement with the experimental data confirming the growth associated nature of PHB production. The best method for recovery of PHB was chemical digestion using sodium hypochlorite alone. The characterization of the produced polymer was carried out using FT-IR, 1HNMR spectroscopy, gel permeation chromatography and transmission electron microscope. The analysis of the nucleotide sequences of PHA synthase enzyme revealed class III identity. The putative tertiary structure of PHA synthase enzyme was analyzed using Modular Approach to Structural class prediction software, Tied Mixture Hidden Markov Model server, and Swiss model software. It was deduced that PHA synthases' structural class was multidomain protein (α/ß) containing a conserved cysteine residue and lipase box as characteristic features of α/ß hydrolase super family. Taken together, all the results of molecular characterization and transmission electron microscope images supported that the PHB formation was attained by the micelle model. To the best of our knowledge, this is the first report on production of growth associated PHB polymer using A. macrocytogenes isolate KC685000, and its class III PHA synthase.


Subject(s)
3-Hydroxybutyric Acid/biosynthesis , 3-Hydroxybutyric Acid/isolation & purification , Pseudomonadaceae/metabolism , 3-Hydroxybutyric Acid/metabolism , Base Sequence , Kinetics , Polymers , Pseudomonadaceae/genetics , Spectroscopy, Fourier Transform Infrared/methods
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