Halotolerant Rhizobacterial Strains Mitigate the Adverse Effects of NaCl Stress in Soybean Seedlings.

BACKGROUND: Salinity is one of the major abiotic constraints that hinder health and quality of crops. Conversely, halotolerant plant growth-promoting rhizospheric (PGPR) bacteria are considered biologically safe for alleviating salinity stress. RESULTS: We isolated halotolerant PGPR strains from the rhizospheric soil of Artemisia princeps, Chenopodium ficifolium, Echinochloa crus-galli, and Oenothera biennis plants; overall, 126 strains were isolated. The plant growth-promoting traits of these isolates were studied by inoculating them with the soil used to grow soybean plants under normal and salt stress (NaCl; 200 mM) conditions. The isolates identified as positive for growth-promoting activities were subjected to molecular identification. Out of 126 isolates, five strains-Arthrobacter woluwensis (AK1), Microbacterium oxydans (AK2), Arthrobacter aurescens (AK3), Bacillus megaterium (AK4), and Bacillus aryabhattai (AK5)-were identified to be highly tolerant to salt stress and demonstrated several plant growth-promoting traits like increased production of indole-3-acetic acid (IAA), gibberellin (GA), and siderophores and increased phosphate solubilization. These strains were inoculated in the soil of soybean plants grown under salt stress (NaCl; 200 mM) and various physiological and morphological parameters of plants were studied. The results showed that the microbial inoculation elevated the antioxidant (SOD and GSH) level and K+ uptake and reduced the Na+ ion concentration. Moreover, inoculation of these microbes significantly lowered the ABA level and increased plant growth attributes and chlorophyll content in soybean plants under 200 mM NaCl stress. The salt-tolerant gene GmST1 was highly expressed with the highest expression of 42.85% in AK1-treated plants, whereas the lowest expression observed was 13.46% in AK5-treated plants. Similarly, expression of the IAA regulating gene GmLAX3 was highly depleted in salt-stressed plants by 38.92%, which was upregulated from 11.26% to 43.13% upon inoculation with the microorganism. CONCLUSION: Our results showed that the salt stress-resistant microorganism used in these experiments could be a potential biofertilizer to mitigate the detrimental effects of salt stress in plants via regulation of phytohormones and gene expression.

Aerobic degradation of 3,3',4,4'-tetrachlorobiphenyl by a resuscitated strain Castellaniella sp. SPC4: Kinetics model and pathway for biodegradation.

Resuscitated strains which were obtained by addition of resuscitation promoting factor (Rpf) could provide a vast majority of microbial source for obtaining highly efficient polychlorinated biphenyl (PCB)-degrading bacteria. In this study, the Castellaniella sp. strain SPC4 which was resuscitated by Rpf addition showed the highest efficiency in degradation of 3,3',4,4'-tetrachlorobiphenyl (PCB 77) among the resuscitated and non-resuscitated isolates. Further investigations on the PCB 77 degradation capability of the resuscitated strain SPC4 showed that SPC4 could efficiently degrade PCB 77 with maximum degradation rate (qmax) of 0.066/h at about 20 mg/L of PCB 77. The maximum growth rate on PCB 77 was 2.663 × 107 CFU/(mL·h) (0.024/h). The most suitable model of Edward demonstrated that the SPC4 could achieve qmax of 0.9315/h, with substrate-affinity of 11.33 mg/L and substrate-inhibition constants of 11.41 mg/L. Meanwhile, the presence of bphA gene expression and chlorine ions release, together with the identification of metabolites, confirmed that the bph-encoded biphenyl pathway was involved in PCB 77 mineralization by SPC4. This report is the first to demonstrate aerobic degradation of PCB 77 by the resuscitated strain Castellaniella sp. SPC4, indicating excellent potential for PCB bioremediation.

Tepidicella baoligensis sp. nov., A Novel Member of Betaproteobacterium Isolated from an Oil Reservoir.

A Gram-negative, non-pigmented, aerobic bacterium, designated strain B18-50T was isolated from oil-well production water in Baolige oilfield, China. The strain was able to grow at pH 6.5-10.5 (optimum at pH 7.5-8.5), in 0-3% (w/v) NaCl (optimum at 0-0.5%, w/v) and at 20-60 °C (optimum at 45 °C). Cells of the isolate were motile with a single polar flagellum and non-spore-forming rods. Organic acids and amino acids were used as carbon and energy sources, but sugars and polyols were not assimilated. The major cellular fatty acids were C16:0, C16:1ω6c/ω7c, and C18:1ω7c. Ubiquinone 8 was the predominant respiratory quinone. The major polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol. The genomic DNA G+C content of the isolate was 62.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain B18-50T was most closely related to Tepidicella xavieri DSM 19605T (97.5% similarity). Comparative analysis of genotypic and phenotypic features indicate that strain B18-50T represents a novel species of the genus Tepidicella, for which the name Tepidicella baoligensis sp. nov. is proposed. The type strain is B18-50T (= CGMCC 1.13575T = KCTC 62779T).

Silver and Copper Acute Effects on Membrane Proteins and Impact on Photosynthetic and Respiratory Complexes in Bacteria.

Silver (Ag+) and copper (Cu+) ions have been used for centuries in industry, as well as antimicrobial agents in agriculture and health care. Nowadays, Ag+ is also widely used in the field of nanotechnology. Yet, the underlying mechanisms driving toxicity of Ag+ ions in vivo are poorly characterized. It is well known that exposure to excess metal impairs the photosynthetic apparatus of plants and algae. Here, we show that the light-harvesting complex II (LH2) is the primary target of Ag+ and Cu+ exposure in the purple bacterium Rubrivivax gelatinosus Ag+ and Cu+ specifically inactivate the 800-nm absorbing bacteriochlorophyll a (B800), while Ni2+ or Cd2+ treatment had no effect. This was further supported by analyses of CuSO4- or AgNO3-treated membrane proteins. Indeed, this treatment induced changes in the LH2 absorption spectrum related to the disruption of the interaction of B800 molecules with the LH2 protein. This caused the release of B800 molecules and subsequently impacted the spectral properties of the carotenoids within the 850-nm absorbing LH2. Moreover, previous studies have suggested that Ag+ can affect the respiratory chain in mitochondria and bacteria. Our data demonstrated that exposure to Ag+, both in vivo and in vitro, caused a decrease of cytochrome c oxidase and succinate dehydrogenase activities. Ag+ inhibition of these respiratory complexes was also observed in Escherichia coli, but not in Bacillus subtilisIMPORTANCE The use of metal ions represents a serious threat to the environment and to all living organisms because of the acute toxicity of these ions. Nowadays, silver nanoparticles are one of the most widely used nanoparticles in various industrial and health applications. The antimicrobial effect of nanoparticles is in part related to the released Ag+ ions and their ability to interact with bacterial membranes. Here, we identify, both in vitro and in vivo, specific targets of Ag+ ions within the membrane of bacteria. This include complexes involved in photosynthesis, but also complexes involved in respiration.

On Burkholderiales order microorganisms and cystic fibrosis in Russia.

BACKGROUND: Microbes infecting cystic fibrosis patients' respiratory tract are important in determining patients' functional status. Representatives of Burkholderiales order are the most dangerous. The goal of our investigation was to reveal the diversity of Burkholderiales, define of their proportion in the microbiome of various parts of respiratory tract and determine the pathogenicity of the main representatives. RESULTS: In more than 500 cystic fibrosis patients, representing all Federal Regions of Russia, 34.0% were infected by Burkholderia cepacia complex (Bcc), 21.0% by Achromobacter spp. and 12.0% by Lautropia mirabilis. B. cenocepacia was the most numerous species among the Bcc (93.0%), and A. ruhlandii was the most numerous among Achromobacter spp. (58.0%). The most abundant genotype in Bcc was sequence type (ST) 709, and in Achromobacter spp. it was ST36. These STs constitute Russian epidemic strains. Whole genome sequencing of strains A. ruhlandii SCCH3:Ach33-1365 ST36 and B. cenocepacia GIMC4560:Bcn122 ST709 revealed huge resistomes and many virulence factors, which may explain the difficulties in eradicating these strains. An experience of less dangerous B. cenocepcia ST710 elimination was described. Massively parallel sequencing of 16S rDNA amplicons, including V1-V4 hypervariable regions, was used to definite "healthy" microbiome characteristics. Analysis of maxillary sinus lavage of 7 patients revealed infection with Proteobacteria of the same ST as pathogens from sputum, suggesting that the maxillary sinus is a source of infection in cystic fibrosis patients. CONCLUSIONS: Characterization of the Russian epidemic bacterial strains in the sputum and sinuses of cystic fibrosis patients have better defined the importance of Burkholderiales bacteria. This information may aid in the development of effective approaches for treatment of this disease.

The biophysics of a critical phenomenon: colonization and sedimentation of the photosynthetic bacteria Rubrivivax gelatinosus.

In response to environmental changes, the photosynthetic bacterium Rubrivivax gelatinosus (Rvx.) can switch from a planktonic lifestyle to a phototrophic biofilm. Like in critical phenomena, the colonization and sedimentation of the cells is abrupt and hard to predict causally, and the underlying biophysics of the mechanisms involved is not known. Herein, we report basic experimental observations and quantitative explanations as keys to understanding microbial turnover of aggregates. (1) The moment of sedimentation can be controlled by the height of the tube of cultivation, by the concentrations of externally added Ficoll (a highly branched polymer) and/or of internally produced polysaccharides (constituents of the biofilm). (2) The observed translational diffusion coefficient of the planktonic bacteria is the sum of diffusion coefficients coming from random Brownian and twitching movements of the bacteria and amounts to 14 (µm)2/s. (3) This value drops hyperbolically with the association number of the cell aggregates and with the concentration of the exopolysaccharides in the biofilm. In the experiments described herein, their effects could be separated. (4) The critical conditions of colonization and sinking of the cells will be achieved if the height of the tube meets the scale height that is proportional to the ratio of the diffusion coefficient and the net mass of the bacterium. The decisive role of the web-like structure of a biofilm, the organization of bacteria from loose cooperativity to solid aggregation, and the possible importance of similar controls in other phototrophic microorganisms are discussed.

Temperate phages as self-replicating weapons in bacterial competition.

Microbial communities are accompanied by a diverse array of viruses. Through infections of abundant microbes, these viruses have the potential to mediate competition within the community, effectively weakening competitive interactions and promoting coexistence. This is of particular relevance for host-associated microbial communities, because the diversity of the microbiota has been linked to host health and functioning. Here, we study the interaction between two key members of the microbiota of the freshwater metazoan Hydra vulgaris The two commensal bacteria Curvibacter sp. and Duganella sp. protect their host from fungal infections, but only if both of them are present. Coexistence of the two bacteria is thus beneficial for Hydra Intriguingly, Duganella sp. appears to be the superior competitor in vitro due to its higher growth rate when both bacteria are grown separately, but in co-culture the outcome of competition depends on the relative initial abundances of the two species. The presence of an inducible prophage in the Curvibacter sp. genome, which is able to lytically infect Duganella sp., led us to hypothesize that the phage modulates the interaction between these two key members of the Hydra microbiota. Using a mathematical model, we show that the interplay of the lysogenic life cycle of the Curvibacter phage and the lytic life cycle on Duganella sp. can explain the observed complex competitive interaction between the two bacteria. Our results highlight the importance of taking lysogeny into account for understanding microbe-virus interactions and show the complex role phages can play in promoting coexistence of their bacterial hosts.

Characterizing bacterial communities in paper production-troublemakers revealed.

Biofilm formation is a major cause of reduced paper quality and increased down time during paper manufacturing. This study uses Illumina next-generation sequencing to identify the microbial populations causing quality issues due to their presence in biofilms and slimes. The paper defects investigated contained traces of the films and/or slime of mainly two genera, Tepidimonas and Chryseobacterium. The Tepidimonas spp. found contributed on average 68% to the total bacterial population. Both genera have been described previously to be associated with biofilms in paper mills. There was indication that Tepidimonas spp. were present as compact biofilm in the head box of one paper machine and was filtered out by the paper web during production. On the other hand Tepidimonas spp. were also present to a large extent in the press and white waters of two nonproblematic paper machines. Therefore, the mere presence of a known biofilm producer alone is not sufficient to cause slimes and therefore paper defects and other critical factors are additionally at play. For instance, we identified Acidovorax sp., which is an early colonizer of paper machines, exhibiting the ability to form extracellular DNA matrices for attachment and biofilm formation.

Ca. Branchiomonas cysticola, Ca. Piscichlamydia salmonis and Salmon Gill Pox Virus transmit horizontally in Atlantic salmon held in fresh water.

Elucidation of the role of infectious agents putatively involved in gill disease is commonly hampered by the lack of culture systems for these organisms. In this study, a farmed population of Atlantic salmon pre-smolts, displaying proliferative gill disease with associated Candidatus Branchiomonas cysticola, Ca. Piscichlamydia salmonis and Atlantic salmon gill pox virus (SGPV) infections, was identified. A subpopulation of the diseased fish was used as a source of waterborne infection towards a population of naïve Atlantic salmon pre-smolts. Ca. B. cysticola infection became established in exposed naïve fish at high prevalence within the first month of exposure and the bacterial load increased over the study period. Ca. P. salmonis and SGPV infections were identified only at low prevalence in exposed fish during the trial. Although clinically healthy, at termination of the trial the exposed, naïve fish displayed histologically visible pathological changes typified by epithelial hyperplasia and subepithelial inflammation with associated bacterial inclusions, confirmed by fluorescent in situ hybridization to contain Ca. B. cysticola. The results strongly suggest that Ca. B. cysticola infections transmit directly from fish to fish and that the bacterium is directly associated with the pathological changes observed in the exposed, previously naïve fish.

Comparison of biofilm formation and motility processes in arsenic-resistant Thiomonas spp. strains revealed divergent response to arsenite.

Bacteria of the genus Thiomonas are found ubiquitously in arsenic contaminated waters such as acid mine drainage (AMD), where they contribute to the precipitation and the natural bioremediation of arsenic. In these environments, these bacteria have developed a large range of resistance strategies among which the capacity to form particular biofilm structures. The biofilm formation is one of the most ubiquitous adaptive response observed in prokaryotes to various stresses, such as those induced in the presence of toxic compounds. This study focused on the process of biofilm formation in three Thiomonas strains (CB1, CB2 and CB3) isolated from the same AMD. The results obtained here show that these bacteria are all capable of forming biofilms, but the architecture and the kinetics of formation of these biofilms differ depending on whether arsenite is present in the environment and from one strain to another. Indeed, two strains favoured biofilm formation, whereas one favoured motility in the presence of arsenite. To identify the underlying mechanisms, the patterns of expression of some genes possibly involved in the process of biofilm formation were investigated in Thiomonas sp. CB2 in the presence and absence of arsenite, using a transcriptomic approach (RNA-seq). The findings obtained here shed interesting light on how the formation of biofilms, and the motility processes contribute to the adaptation of Thiomonas strains to extreme environments.