Genome analysis uncovers the prolific antagonistic and plant growth-promoting potential of endophyte Bacillus velezensis K1.

Insights into the application of endophytic bacilli in sustainable agricultural practices have opened up new avenues for the inhibition of soil-borne pathogens and the improvement of plant health. Bacillus subtilis K1, an endophytic bacterium originally isolated from aerial roots of Ficus benghalensis is a potential biocontrol agent secreting a mixture of surfactins, iturins and fengycins. The current study extends the characterization of this bacterium through genomic and comparative genomics approaches. The sequencing of the bacterial genome at Illumina MiSeq platform revealed that it possessed a 4,103,502-bp circular chromosome with 45.98% GC content and 4325 predicted protein-coding sequences. Based on phylogenomics and whole-genome average nucleotide identity, the B. subtilis K1 was taxonomically classified as Bacillus velezensis. The formerly evaluated phenotypic traits viz. C-source utilization and lipopeptide-mediated fungal antagonism were correlated to their molecular determinants. The genome also harbored several genes associated with induced systemic resistance and plant growth promotion i.e, phytohormone production, nitrogen assimilation and reduction, siderophore production, phosphate solubilization, biofilm formation, swarming motility, acetoin and butanediol synthesis. The production of antifungal volatile organic compounds and plant growth promotion was experimentally demonstrated by volatile compound assay and seed germination assay on cumin and groundnut. The isolate also holds great prospects for application as a soil inoculant as indicated by enhancement in the growth of groundnut via in planta pot studies. Bacterial pan-genome analysis based on a comparison of whole genomes with eighteen other Bacillus strains was also conducted. Comparative examination of biosynthetic gene clusters across all genomes indicated that the largest number of gene clusters were harbored by the K1 genome. Based on the findings, we propose K1 as a model for scrutinizing non-ribosomally synthesized peptide synthetase and polyketide synthetase derived molecules.

Transcriptome profiling reveals upregulation of benzoate degradation and related genes in Pseudomonas aeruginosa D6 during textile dye degradation.

An upsurge in textile dye pollution has demanded immediate efforts to develop an optimum technology for their bioremediation. However, the molecular mechanism underpinning aerobic decolorization of dyes is still in its infancy. Thus, in the current work, the intricacies of aerobic remediation of textile dyes by Pseudomonas aeruginosa D6 were understood via a transcriptomic approach. The bacterium isolated from the sludge sample of a common effluent treatment plant was able to decolorize 54.42, 57.66, 50.84 and 65.86% of 100 mg L-1 of four different dyes i.e., TD01, TD04, TD05, and TD06, respectively. The maximum decolorization was achieved within six days and thus, the first and sixth day of incubation were selected for transcriptome analysis at the early and late phase of the decolorization, respectively. The expression profiles of all samples were compared to gain insight into the dye-specific response of bacterium and it was found that it behaved most uniquely in the presence of the dye TD01. Several genes critical to core metabolic processes like the TCA cycle, glycolysis, pentose phosphate pathway, translation, cell motility etc. Were found to be overexpressed in the presence of dyes. Interestingly, in response to dyes, the benzoate degradation pathway was significantly upregulated in the bacterium as compared to control (i.e., bacterium without dye). Thus, seven genes contributing to the induction of the same were further studied by RT-qPCR analysis. Overall, the involvement of the benzoate pathway implies the appearance of aromatic intermediates during decolorization, which in turn infers dye degradation.

Genome analysis to decipher syntrophy in the bacterial consortium 'SCP' for azo dye degradation.

BACKGROUND: A bacterial consortium SCP comprising three bacterial members, viz. Stenotrophomonas acidaminiphila APG1, Pseudomonas stutzeri APG2 and Cellulomonas sp. APG4 was developed for degradation of the mono-azo dye, Reactive Blue 28. The genomic analysis of each member of the SCP consortium was done to elucidate the catabolic potential and role of the individual organism in dye degradation. RESULTS: The genes for glycerol utilization were detected in the genomes of APG2 and APG4, which corroborated with their ability to grow on a minimal medium containing glycerol as the sole co-substrate. The genes for azoreductase were identified in the genomes of APG2 and APG4, while no such trait could be determined in APG1. In addition to co-substrate oxidation and dye reduction, several other cellular functions like chemotaxis, signal transduction, stress-tolerance, repair mechanisms, aromatic degradation, and copper tolerance associated with dye degradation were also annotated. A model for azo dye degradation is postulated, representing the predominant role of APG4 and APG2 in dye metabolism while suggesting an accessory role of APG1. CONCLUSIONS: This exploratory study is the first-ever attempt to divulge the genetic basis of azo-dye co-metabolism by cross-genome comparisons and can be harnessed as an example for demonstrating microbial syntrophy.

Genome analysis reveals probiotic propensities of Paenibacillus polymyxa HK4.

The legislations on the usage of antibiotics as growth promoters and prophylactic agents have compelled to develop alternative tools to upsurge the animal protection and contain antibiotic usage. Probiotics have emerged as an effective antibiotic substitute in animal farming. The present study explores the probiotic perspective of Paenibacillus polymyxa HK4 interlinking the genotypic and phenotypic characteristics. The draft genome of HK4 revealed the presence of ORFs encoding the functions associated with tolerance to gastrointestinal stress and adhesion. The biosynthetic gene clusters encoding non-ribosomally synthesized peptides, polyketides and lanthipeptides such as fusaricidin, tridecaptin, polymyxin, paenilan and paenibacillin were annotated in HK4 genome. The strain harbored the chromosomal gene conferring the resistance to lincosamides. No functional gene encoding virulence or toxins could be identified in the genome of HK4. The genome analysis data was complemented by the in vitro experiments confirming its survival during gastrointestinal transit, antimicrobial potential and antibiotic sensitivity. NUCLEOTIDE SEQUENCE ACCESSION NUMBER: The draft-genome sequence of Paenibacillus polymyxa HK4 has been deposited as whole-genome shotgun project at GenBank under the accession number PRJNA603023.

Decoding social behaviors in a glycerol dependent bacterial consortium during Reactive Blue 28 degradation.

Biodegradation of reactive azo dyes has been an arduous problem for decades. Several efficient biosystems have been proposed for dye degradation, but most of them are dependent on the availability of costly co-substrates such as peptone, yeast extract, and/or glucose. The present study describes the azo dye degradation by a bacterial consortium using glycerol as a sole co-substrate. The consortium was developed from a mixed bacterial culture obtained upon enrichment of soil sediment for Reactive Blue 28 (RB28) decolorization in the presence of glycerol (0.1%; v/v). The consortium with three bacterial species, i.e., Stenotrophomonas acidaminiphila APG1, Cellulomonas sp. APG4, and Pseudomonas stutzeri APG2, designated as "SCP," decolorized 92% of 100 ppm dye in 96 h. The intricacies of the interactions existing within the members of the consortium were resolved by a simple and unique analysis called "BSocial." Among all the members, Cellulomonas sp. APG4 exerted a net-positive impact for decolorization (%) on the consortium. The net fitness of the community increased when all the three species were present, and thus, all of them were selected for further analysis. Moreover, APG4 seemed to be central in the reductive decolorization as it possessed the highest reductase activity. The dye degradation by the consortium was demonstrated by UV-Visible spectroscopy, HPTLC, and FTIR spectroscopy of control and decolorized cell-free supernatant. The LC-ESI-MS analysis of metabolites extracted from decolorized cell-free medium led to the identification of degradation products, thus leading us to propose the plausible pathway for degradation of RB28 by bacterial consortium.

Characterization of an extremely halotolerant Staphylococcus arlettae HPSSN35C isolated from Dwarka Beach, India.

An extremely halotolerant bacterium designated as HPSSN35C was isolated from saline soil of Dwarka beach, India. It exhibited growth over a wide range of NaCl in medium varying from 0 to 6 M. The isolate produced peach-pink pigment above ∼1.3 M NaCl. The culture was characterized using biochemical tests, bioMerieux Staph identification kit, API ID32 Staph system, and Biolog. Due to slow growth and extreme salt tolerance no ID was obtained in Biolog. Antibiotic sensitivity to various antibiotics was tested. Phenotypic characterization showed that it belonged to the novobiocin resistant staphylococci group. Analysis of 16S rRNA gene sequence comparison of 1452 base pairs showed that isolate is closely related to Staphylococcus saprophyticus group with close relationship to Staphylococcus arlettae (99% similarity). The halotolerant S. arlettae described in literature till date have been reported to tolerate 4.5 M NaCl and produce white to yellow pigment. The present study reports for the first time extremely halotolerant S. arlettae exhibiting growth up to ∼6 M NaCl and producing peach-pink pigment above ∼1.3 M NaCl.