Integrated genomics and proteomics analysis of Paenibacillus peoriae IBSD35 and insights into its antimicrobial characteristics.

Antimicrobial resistance has been developing fast and incurring a loss of human life, and there is a need for new antimicrobial agents. Naturally occurring antimicrobial peptides offer the characteristics to counter AMR because the resistance development is low or no resistance. Antimicrobial peptides from Paenibacillus peoriae IBSD35 cell-free supernatant were salted out and purified using chromatography and characterized with liquid chromatography-tandem-mass spectrometry. The extract has shown a high and broad spectrum of antimicrobial activity. Combining the strain IBSD35 genome sequence with its proteomic data enabled the prediction of biosynthetic gene clusters by connecting the peptide from LC-MS/MS data to the gene that encode. Antimicrobial peptide databases offered a platform for the effective search, prediction, and design of AMPs and expanded the studies on their isolation, structure elucidation, biological evaluation, and pathway engineering. The genome-based taxonomy and comparisons have shown that P. peoriae IBSD35 is closely related to Paenibacillus peoriae FSL J3-0120. P. peoriae IBSD35 harbored endophytic trait genes and nonribosomal peptide synthases biosynthetic gene clusters. The comparative genomics revealed evolutionary insights and facilitated the discovery of novel SMs using proteomics from the extract of P. peoriae IBSD35. It will increase the potential to find novel bio-molecules to counter AMR.

Draft genome sequence and functional analysis of Lysinibacillus xylanilyticus t26, a plant growth-promoting bacterium isolated from Capsicum chinense rhizosphere.

Capsicum chinense is the chilli species containing the highest amount of capsaicin, and is an important traditional spice crop of Northeast India. Capsaicinoids derived from C. chinense are used in anticancer and anti-obesity treatments, as temperature regulators, in pain therapy, and as antioxidants. The current production and yield are very low due to the lack of organized cultivation and scientific inputs, and various plant diseases. Synthetic pesticides are frequently applied to boost yields, which creates potential risks to the environment, crops, and humans. The use of plant growth-promoting rhizobacteria is an alternative strategy in crop disease management to reduce the dependency on agrochemicals, which have detrimental effects on the environment. Lysinibacillus xylanilyticus t26 isolated from the C. chinense rhizosphere has shown good prospects in plant growth promotion and biocontrol. It showed strong antagonistic activity against Pythium ultimum ITCC 1650, Rhizoctonia solani ITCC 6491, and Fusarium oxysporum ITCC 6246. The draft genome sequencing of L. xylanilyticus t26 yielded a total of 5.69 Mbp with a G+C content of 36.80%. Genome analysis revealed that L. xylanilyticus t26 is very similar to L. xylanilyticus MH683160.1, and is phylogenetically related to L. xylanilyticus IBBPo7. Bioinformatics analysis predicted that it harbored type III polyketides, non-ribosomal peptides, terpenes, and lantibiotics including cerecidin, bacteriocins, siderophores, and thiopeptides, which are important traits of rhizobacteria for the utilization of minerals and to compete with other microbes for food. The strain t26 is a potential biocontrol agent for soil-borne fungal diseases. In this study, we derived the possible siderophore production pathways through the analysis of L. xylanilyticus t26 draft genome and plant growth response bioassays. The availability of genome data provides information that this draft genome harbored a siderophore BGC, which is 33% similar to petrobactin.

Bioethanol production from Ficus fruits (Ficus cunia) by Fusarium oxysporum through consolidated bioprocessing system.

Fusarium oxysporum is among the few filamentous fungi capable of fermenting ethanol directly from lignocellulose biomass (LCB). It has the essential enzymatic toolbox to disintegrate LCB to its monosaccharides, which subsequently fermented to ethanol under anaerobic and micro-aerobic conditions. However, the structural complexity of LCB and modest performances of wild fungi are major limitations for application in local biorefineries. This study assessed the potential of the locally isolated Fusarium oxysporum for the production of bioethanol from Ficus fruits (Ficus cunia) using Consolidated Bioprocessing (CBP). The maximum ethanol concentration achieved was at 5% substrate loadings with pH 6 irrespective of temperature variance, attaining a concentration of 3.54 g/L and 3.88 g/L at 28 °C and 32 °C, respectively. The monitoring of analytes (glucose, arabinose, cellobiose, xylose, acetic acid, ethanol, furfural, and HMF) in this study suggests the utilization of an array of sugars released from Ficus fruits, irrespective of the difference in the process parameters. This study also shows that CBP of freshly grounded Ficus fruits was feasible employing a mild hydrothermal pretreatment (autoclaved at 121 °C for 30 min in 1:10 w/v) and without supplementing any extraneous enzymes. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03234-y.

Bacterial rhizosphere community profile at different growth stages of Umorok (Capsicum chinense) and its response to the root exudates.

The role of microflora is an indispensable part of the living organisms. Plants actively recruit specific microbial community to establish favorable habitat with the distinct microbiome, essentially unique for each species, offering new opportunities for plant growth and productivity. Umorok, an indigenous chili variety of northeastern India, production is highly affected by various factors; therefore, rhizosphere bacteria and their relationship with the root exudates released were analyzed to demonstrate rhizosphere bacterial impact on plant growth and health. Culturable and metagenomic bacterial DNA was characterized and the chemical nature of the root exudate was analyzed using chemotaxis assay after its basic analysis in HPLC. Juvenile stage exhibited diverse bacterial species of gammaproteobacteria, alphaproteobacteria, and actinobacteria but lacked the betaproteobacteria while the microbial diversity was reduced in flowering and fruiting stages. However, every growth stage maintained a similar amount of bacterial population regardless of diversity. The population of Pseudomonas, Bacillus, and Burkholderia species was increased several folds in flowering and fruiting stage. Further, the chemotaxis assay unveiled the advantage of root exudate chemical composition for specific microbial recruitment. The chemical composition analysis of root exudates showed substantial variation in the concentration of organic acids, phenolics, and flavonoids that are favoring unique bacterial species. Thus, root exudates confer and limit the related microbial population besides typical plant-bacterial synergetic association. This study emphasized information about the type of microbial load present in each growth stage, which is essential to develop a microbial consortia package for Umorok overall crop improvement.

Bio-mining the forest ecosystem of North East India for identification of antimicrobial metabolites from fungi through submerged fermentation.

In this study, fungi isolated from less explored forest soil ecosystem of Northeast India were studied for the production of potential antimicrobial metabolites (AMM). Out of the 68 fungi isolated from forest soil of Manipur, 7 of them showed AMA against the test pathogens. Among them, Aspergillus terreus (IBSD-F4) showed the most significant activity against Staphylococcus aureus (ATCC-25923), Bacillus anthracis (IBSD-C370), Pseudomonas fluorescens (ATCC-13525), Salmonella typhimurium (ATCC-14028), Escherichia coli (ATCC-25922) and Candida albicans (ATCC-10231). The active metabolite was harvested from the fermentation broth of Aspergillus terreus and purified by column chromatography and semi preparative-HPLC. The compound was identified as 'Sclerotionigrin A' on the basis of UV-vis spectra, MS and NMR analyses. This compound was reported for the first time from A. terreus. The study highlights, the importance of exploring microbes from forest soil for identification of bioactive metabolites for future drug development.

Host-range dynamics of Cochliobolus lunatus: from a biocontrol agent to a severe environmental threat.

We undertook an investigation to advance understanding of the host-range dynamics and biocontrol implications of Cochliobolus lunatus in the past decade. Potato (Solanum tuberosum L) farms were routinely surveyed for brown-to-black leaf spot disease caused by C. lunatus. A biphasic gene data set was assembled and databases were mined for reported hosts of C. lunatus in the last decade. The placement of five virulent strains of C. lunatus causing foliar necrosis of potato was studied with microscopic and phylogenetic tools. Analysis of morphology showed intraspecific variations in stromatic tissues among the virulent strains causing foliar necrosis of potato. A maximum likelihood inference based on GPDH locus separated C. lunatus strains into subclusters and revealed the emergence of unclustered strains. The evolving nutritional requirement of C. lunatus in the last decade is exhibited by the invasion of vertebrates, invertebrates, dicots, and monocots. Our results contribute towards a better understanding of the host-range dynamics of C. lunatus and provide useful implications on the threat posed to the environment when C. lunatus is used as a mycoherbicide.

Secretome weaponries of Cochliobolus lunatus interacting with potato leaf at different temperature regimes reveal a CL[xxxx]LHM - motif.

BACKGROUND: Plant and animal pathogenic fungus Cochliobolus lunatus cause great economic damages worldwide every year. C. lunatus displays an increased temperature dependent-virulence to a wide range of hosts. Nonetheless, this phenomenon is poorly understood due to lack of insights on the coordinated secretome weaponries produced by C. lunatus under heat-stress conditions on putative hosts. To understand the mechanism better, we dissected the secretome of C. lunatus interacting with potato (Solanum tuberosum L.) leaf at different temperature regimes. RESULTS: C. lunatus produced melanized colonizing hyphae in and on potato leaf, finely modulated the ambient pH as a function of temperature and secreted diverse set of proteins. Using two dimensional gel electrophoresis (2-D) and mass spectrometry (MS) technology, we observed discrete secretomes at 20°C, 28°C and 38°C. A total of 21 differentially expressed peptide spots and 10 unique peptide spots (that did not align on the gels) matched with 28 unique protein models predicted from C. lunatus m118 v.2 genome peptides. Furthermore, C. lunatus secreted peptides via classical and non-classical pathways related to virulence, proteolysis, nucleic acid metabolism, carbohydrate metabolism, heat stress, signal trafficking and some with unidentified catalytic domains. CONCLUSIONS: We have identified a set of 5 soluble candidate effectors of unknown function from C. lunatus secretome weaponries against potato crop at different temperature regimes. Our findings demonstrate that C. lunatus has a repertoire of signature secretome which mediates thermo-pathogenicity and share a leucine rich "CL[xxxx]LHM"-motif. Considering the rapidly evolving temperature dependent-virulence and host diversity of C. lunatus, this data will be useful for designing new protection strategies.

Grappling the high altitude for safe edible bamboo shoots with rich nutritional attributes and escaping cyanogenic toxicity.

Consumption of bamboo species with high level of total cyanogenic content (TCC) in Asia by many ethnic groups is significantly associated with food poisoning and occasionally Konzo (a neurological disorder). Adequate characterization of edible bamboo species with low level of TCC and high nutritious attributes is required for consumer's safety as well as for the conservation of the gene pool. Here, we employed morphological descriptors, atomic absorption spectrophotometer, RAPD, and trnL-F intergenic spacer to characterize 15 indigenous edible bamboo species of north-east India. The study indicates that morphologically and genetically evolved edible bamboo species having large and robust bamboo-shoot texture and growing at low altitude contain high level of TCC, low antioxidant properties, and low levels of beneficial macronutrients and micronutrients. Importantly, Dendrocalamus species are shown to be rich in TCC irrespective of the growing altitude while Bambusa species are found to have moderate level of TCC. The findings clearly demonstrated that Chimonobambusa callosa growing at high altitude represents safe edible bamboo species with nutritious attributes.