Differential RNA-seq of Plant Beneficial Rhizobacterium Bacillus amyloliquefaciens FZB42 Reveals sRNA Bas01 involved in Sporulation and Biofilm Formation
ABSTRACT: Bacillus amyloliquefaciens FZB42 is a representative organism for Gram positive soil bacteria associated with plant roots and beneficial to plant growth. It is of immense importance to understand mechanisms of this class of bacteria adapting to rhizosphere. In this work employing differential RNA sequencing (RNA-seq) and Northern blot, we systematically identified transcription start sites of mRNAs as well as non-coding regulatory RNAs in FZB42. The genes regulated at different growth phases and located in polycistronic operons were also identified. A set of genes were re-annotated. In addition, a sRNA named Bas01 was identified to be involved in Bacillus sporulation and biofilm formation. The result we obtained provides valuable data for investigation of Bacillus gene expression and molecular details of rhizobacterial interaction with host plants. Examination of transcriptome profile of rhizobacterium B. amyloliquefaciens FZB42 grown under six conditions.
Bacillus amyloliquefaciens subsp. plantarum FZB42 is a representative of Gram-positive plant-growth-promoting rhizobacteria (PGPR) that inhabit plant root environments. In order to better understand the molecular mechanisms of bacteria-plant symbiosis, we have systematically analyzed the primary transcriptome of strain FZB42 grown under rhizosphere-mimicking conditions using differential RNA sequencing (dRNA-seq). Our analysis revealed 4,877 transcription start sites for protein-coding genes, id ...[more]
Project description:In bacteria and archaea, CRISPR loci confer adaptive, sequence-based immunity against viruses and plasmids. CRISPR interference is specified by CRISPR RNAs (crRNAs) that are transcribed and processed from CRISPR spacers and repeats. Pre-crRNA processing is essential for CRISPR interference in all systems studied thus far. Here we examine crRNA biogenesis and CRISPR interference in naturally competent Neisseria spp., including the human pathogen N. meningitidis. Our studies reveal a unique crRNA maturation pathway in which crRNA transcription is driven by promoters that are embedded within each repeat, yielding crRNA 5’ ends are not formed by processing. Although crRNA 3’ end formation occurs through RNase III cleavage of a pre-crRNA/tracrRNA duplex, as in other Type II CRISPR systems, this processing event is dispensable for interference. The meningococcal pathway is the most streamlined CRISPR/cas system characterized to date. Endogenous CRISPR spacers frequently target genomic sequences of other Neisseria strains and so limit natural transformation, which is the primary source of genetic variation that contributes to immune evasion, antibiotic resistance, and virulence in N. meningitidis. dRNA-seq approach for RNA samples from cultures of N. lactamica 020-06, harvested at mid-log. Two cDNA libraries from total RNA were prepared to distinguish between transcripts with either primary orprocessed 5’ ends: one library is generated from untreated RNA, whereas the other is treated with terminator exonuclease (TEX),
Project description:Transcription profiling by array of Bacillus amyloliquefaciens strain FZB42 after interaction exudates (IE) treatment, at OD600=1.0 and at OD600=1.0 respectively. IE was the root exudates prepared from maize plants growing with FZB42. The reference was treated with the root exudates (RE), prepared from maize plants grown in an axenic system.
Project description:Bacillus amyloliquefaciens FZB42 is a well-studied Gram-positive plant growth-promoting rhizobacteria. In this work we aim to study the effect of the effect of sigD deletion on the transcriptome of FZB42. The transcritomes were compared by two-color microarray of the sigD- mutant and the wildtype of B. amyloliquefaciens FZB42 grown in 1C medium supplemented with soil extract (SE). This submission includes data from two independent experiments with three biological replicates. Here a biological replicate means the bacterial culture from one flask used for RNA preparation. The samples were collected by two performers. The experiments were varied in sample performer, the date of the experiment.
Project description:The effect of maize seed (Zea mays L. var. Surprise) exudates on Bacillus amyloliquefaciens FZB42 cultures grown to OD600 = 1.0 or 3.0 was tested against a negative seed exudate control sample.
Project description:Transcriptional profiling by array of Bacillus amyloliquefaciens FZB42 at log phase(OD600=1.0) and early stationary phase(OD600=3.0) after maize root exudate treatment of three concentrations(0.25mg/ml,0.50mg/ml,1.00mg/ml).
Project description:The transcriptomic response of Bacillus amyloliquefaciens FZB42 to maize root exudates at OD600=3.0. This is a comprehensive analysis using the data of six microarray experiments (Exp1-2-3 and ExpABC respectively, 18 hybridization in total).
Project description:Bacteria possess many small noncoding RNAs whose regulatory roles in pathogenesis are little understood due to a paucity of macroscopic phenotypes in standard virulence assays. Here, we use a novel Dual RNA-seq approach for a single-step simultaneous RNA profiling in both pathogen and host to reveal molecular phenotypes of sRNAs during infection with Salmonella Typhimurium. We identify a new PhoP/Q-activated small RNA which upon bacterial internalization acts to temporally control the expression of both, invasion-associated effectors and virulence genes required for intracellular survival. This riboregulatory activity is shown to adjust the human response to replicating Salmonella, and have a pervasive impact on host RNA expression both inside and outside protein-coding regions including infection-specific alterations of an array of long noncoding RNAs. Our study provides a paradigm for a comprehensive RNA-based analysis of intracellular bacterial pathogens without their physical purification from a host and a new discovery route for hidden functions of pathogen genes. Comparative RNA-seq (in vitro medium shift experiment)
Project description:Bacillus amyloliquefaciens FZB42 is a well-studied Gram-positive plant growth-promoting rhizobacteria. In this work we aim to study the effect of the effect of sigD deletion on the transcriptome of FZB42. The transcritomes were compared by two-color microarray of the sigD- mutant and the wildtype of B. amyloliquefaciens FZB42 grown in 1C medium supplemented with soil extract (SE) and in 1C medium supplemented with both SE and maize root exudates (RE). This submission includes data from three independent experiments with three biological replicates. Here a biological replicate means the bacterial culture from one flask used for RNA preparation. The samples were collected by two performers. The experiments were varied in sample performer, the date of the experiment.