Lichens are complex symbiotic association of mycobionts, photobionts, and bacteriobionts, including chemolithotropic bacteria. In the present study, 46 lichenized bacteria were isolated by conventional and enrichment culture methods on nitrogen-free bromothymol blue (NFb) medium. Only 11 of the 46 isolates fixed nitrogen on NFb and had reduced acetylene. All these 11 isolates had also produced siderophore and 10 of them the IAA. Further, ammonia production was recorded from nine of these nitrogen fixers (NF). On molecular characterization, 16 S rRNA sequencing recorded that, nine NF belonged to Proteobacteria, within Gammaproteobacteria, and were closely related to Enterobacter sp. with a maximum similarity to Enterobacter cloacae. Each one of our NF isolates was aligned closely to Enterobacter pulveris strain E443, Cronobacter sakazakii strain PNP8 and Providencia rettgeri strain ALK058. Notably, a few strains we examined found to possess plant growth promoting properties. This is the first report of Enterobacter sp. from lichens which may be inhabit lichen thalli extrinsically or intrinsically.
Background
Celiac disease is an intestinal chronic disorder with multifactorial etiology resulting in small intestinal mucosal injuries and malabsorption. In genetically predisposed individuals with HLA DQ2/DQ8 molecules, the gluten domains rich in glutamine and proline present gluten domains to gluten reactive CD4+ T cells causing injury to the intestine. In the present experimental design, the indigenous bacteria from wheat samples were studied for their gluten hydrolyzing functionality.
Results
Proteolytic activity of Bacillus spp. was confirmed spectrophotometrically and studied extensively on gliadin-derived synthetic enzymatic substrates, natural gliadin mixture, and synthetic highly immunogenic 33-mer peptide. The degradation of 33-mer peptide and the cleavage specificities of the selected isolates were analyzed by tandem mass spectrometry. The gluten content of the sourdough fermented by the chosen bacterial isolates was determined by R5 antibody based competitive ELISA. All the tested isolates efficiently hydrolyzed Z-YPQ-pNA, Z-QQP-pNA, Z-PPF-pNA, and Z-PFP-pNA and also cleaved 33-mer immunogenic peptide extensively. The gluten content of wheat sourdough was found to be below 110 mg/kg.
Conclusion
It has been inferred that four Bacillus spp especially GS 188 could be useful in developing gluten-reduced wheat food product for celiac disease prone individuals.
Lichens comprise highly diverse and complex microbial communities, the majority consisting of mycobiont, photobiont, Basidiomycetes yeast and bacteriobiont (internal bacterial communities). In this study, bacterial diversity of foliose lichen was reported. Next generation sequence (NGS) such as Illumina Sequencing (150*2) of 16S rRNA (V3 and V6 region) was used to delineate the bacterial communities associated with five foliose lichen samples. Bacterial sequences obtained from lichen samples suggested that, they harboured bacterial community with variable relative abundances. Among all bacterial communities, Alphaproteobacteria were dominant in all the tested lichen samples. The principal coordinate analysis, Venn and bar chart showed significant microbial changes between the different useful bacterial lineages across the lichens. The relative abundance of dominant and rare bacterial species found were varied, diverse, distinct and unique in each lichen. The Proteobacteria 48.19%, Actinobacteria 25.70%, Bacteroidetes 8.53%, Acidobacteria 9.36% and Chloroflexi 0.83% were predominant in all tested lichens. The present empirical study enhances the confirmed knowledge of bacterial diversity inevitably associated with lichens and is the first report on lichenized bacterial diversity and perhaps their potential possible role in lichen symbiosis in addition to phycobiont and mycobiont.
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