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Leptothrix cholodnii SP-6

ABSTRACT: DNA sequencing of wild-type and lthC mutant in Leptothrix cholodnii SP-6

PROVIDER: PRJDB18186 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	DRR568669_1.fastq.gz	Fastqsanger.gz
	DRR568669_2.fastq.gz	Fastqsanger.gz
	DRR568670_1.fastq.gz	Fastqsanger.gz
	DRR568670_2.fastq.gz	Fastqsanger.gz

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Leptothrix cholodnii SP-6

Project description:RNA sequencing of wild-type and lthC mutant in Leptothrix cholodnii SP-6

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Project description:Microorganisms are widely utilized for the treatment of wastewater in activated sludge systems. However, the uncontrolled growth of filamentous bacteria leads to bulking and adversely affects wastewater treatment efficiency. To clarify the nutrient requirements for filament formation, we track the growth of a filamentous bacterium, Leptothrix cholodnii SP-6 in different nutrient-limited conditions using a high aspect-ratio microfluidic chamber to follow cell-chain elongation and sheath formation. We find that limitations in Na+, K+, and Fe2+ yield no observable changes in the elongation of cell chains and sheath formation, whereas limitations of C, N, P, or vitamins lead to more pronounced changes in filament morphology; here we observe the appearance of partially empty filaments with wide intercellular gaps. We observe more dramatic differences when SP-6 cells are transferred to media lacking Mg2+ and Ca2+. Loss of Mg2+ results in cell autolysis, while removal of Ca2+ results in the catastrophic disintegration of the filaments. By simultaneously limiting both carbon and Ca2+ sources, we are able to stimulate planktonic cell generation. These findings paint a detailed picture of the ecophysiology of Leptothrix, which may lead to improved control over the unchecked growth of deleterious filamentous bacteria in water purification systems.

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Project description:Genetic strategies such as gene disruption and fluorescent protein tagging largely contribute to understanding the molecular mechanisms of biological functions in bacteria. However, the methods for gene replacement remain underdeveloped for the filamentous bacteriaLeptothrix cholodniiSP-6. Their cell chains are encased in sheath composed of entangled nanofibrils, which may prevent the conjugation for gene transfer. Here, we describe a protocol optimized for gene disruption through gene transfer mediated by conjugation withEscherichia coliS17-1 with details on cell ratio, sheath removal, and loci validation. The obtained deletion mutants for specific genes can be used to clarify the biological functions of the proteins encoded by the target genes. Graphical overview.

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Project description:In this work, we report that the knockout mutant in other of this luxR-like genes (named luxR402) in Novosphingobium sp. HR1a; floculated faster than the wild-type when cultures are in repose. Transcriptomic analysis allowed us to determine the LuxR402 regulon; we have identified that the carbohydrate assimilation pathway and TCA cycle are affected in the mutant. Accordingly, the mutant presented poorer growth than the wild-type when growing in different carbon sources. At stationary phase of growth the pili biosynthesis, trehalose utilization and capside-related proteins were affected in the mutant strain. Microscopy assays determined that the mutant strain cultures presented cells aggrupations and that the extracellular matrix is less abundant than in the wild-type cultures.

2023-07-13 | GSE223267 | GEO

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