Discovery and profiling of small RNAs responsive to stress conditions in the plant pathogen Pectobacterium atrosepticum
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Date
Authors
Kwenda, Stanford
Gorshkov, Vladimir
Ramesh, Aadi Moolam
Naidoo, Sanushka
Rubagotti, Enrico
Birch, Paul R. J.
Moleleki, Lucy N.
Journal Title
Journal ISSN
Volume Title
Publisher
BioMed Central
Abstract
BACKGROUND : Small RNAs (sRNAs) have emerged as important regulatory molecules and have been studied in
several bacteria. However, to date, there have been no whole-transcriptome studies on sRNAs in any of the Soft Rot
Enterobacteriaceae (SRE) group of pathogens. Although the main ecological niches for these pathogens are plants, a
significant part of their life cycle is undertaken outside their host within adverse soil environment. However, the
mechanisms of SRE adaptation to this harsh nutrient-deficient environment are poorly understood.
RESULTS : In the study reported herein, by using strand-specific RNA-seq analysis and in silico sRNA predictions, we
describe the sRNA pool of Pectobacterium atrosepticum and reveal numerous sRNA candidates, including those that
are induced during starvation-activated stress responses. Consequently, strand-specific RNA-seq enabled detection
of 137 sRNAs and sRNA candidates under starvation conditions; 25 of these sRNAs were predicted for this bacterium in
silico. Functional annotations were computationally assigned to 68 sRNAs. The expression of sRNAs in P. atrosepticum
was compared under growth-promoting and starvation conditions: 68 sRNAs were differentially expressed with 47
sRNAs up-regulated under nutrient-deficient conditions. Conservation analysis using BLAST showed that most of the
identified sRNAs are conserved within the SRE. Subsequently, we identified 9 novel sRNAs within the P. atrosepticum
genome.
CONCLUSIONS : Since many of the identified sRNAs are starvation-induced, the results of our study suggests that sRNAs
play key roles in bacterial adaptive response. Finally, this work provides a basis for future experimental characterization
and validation of sRNAs in plant pathogens.
Description
Additional file 1: Table S1. Complete list of RNA-seq detected sRNAs
(XLSX 19 kb)
Additional file 2: Table S2. Predicted transcription start sites of RNA-seq detected sRNAs (XLSX 9114 kb)
Additional file 3: Table S3. List of in silico predicted sRNAs using RITs from WebGester DB. S2A: Forward strand predictions. S2B: Complementary strand predictions (XLSX 30 kb)
Additional file 4: Table S4. Combined list of predicted sRNA using SIPHT and RITs from WebGester DB. S3A: Matches of in silico predictions with SIPHT (forward strand) S3B: Matches of in silico predictions with SIPHT (complementary strand) (XLSX 15 kb)
Additional file 5: Table S5. Conservation analysis in Soft Rot Enterobacteriaceae (XLSX 16 kb)
Additional file 6: Table S6. Confirmation of RT-PCR amplicons by sequencing and BLASTn against respective sRNA sequences (XLSX 9 kb)
Additional file 7: Table S7. Differentially expressed sRNA under nutrient-rich and starvation conditions (XLSX 19 kb)
Additional file 8: Table S8. List of primers used for RT-PCR validation of novel sRNAs (DOCX 12 kb)
Additional file 9: Table S9. List of primers used for RT-qPCR validation of RNA-seq expression data (DOCX 12 kb)
Additional file 2: Table S2. Predicted transcription start sites of RNA-seq detected sRNAs (XLSX 9114 kb)
Additional file 3: Table S3. List of in silico predicted sRNAs using RITs from WebGester DB. S2A: Forward strand predictions. S2B: Complementary strand predictions (XLSX 30 kb)
Additional file 4: Table S4. Combined list of predicted sRNA using SIPHT and RITs from WebGester DB. S3A: Matches of in silico predictions with SIPHT (forward strand) S3B: Matches of in silico predictions with SIPHT (complementary strand) (XLSX 15 kb)
Additional file 5: Table S5. Conservation analysis in Soft Rot Enterobacteriaceae (XLSX 16 kb)
Additional file 6: Table S6. Confirmation of RT-PCR amplicons by sequencing and BLASTn against respective sRNA sequences (XLSX 9 kb)
Additional file 7: Table S7. Differentially expressed sRNA under nutrient-rich and starvation conditions (XLSX 19 kb)
Additional file 8: Table S8. List of primers used for RT-PCR validation of novel sRNAs (DOCX 12 kb)
Additional file 9: Table S9. List of primers used for RT-qPCR validation of RNA-seq expression data (DOCX 12 kb)
Keywords
Strand-specific RNA-seq, Pectobacterium atrosepticum, In silico prediction, Transcriptome, Riboswitches, 5′ UTR, 3′ UTR, Small RNAs (sRNAs), Soft Rot Enterobacteriaceae (SRE)
Sustainable Development Goals
Citation
Kwenda, S, Gorshkov, V, Ramesh, AM, Naidoo, S, Rubagotti E, Birch, PRJ & Moleleki, LN 2016, 'Discovery and profiling of small RNAs responsive to stress conditions in the plant pathogen Pectobacterium atrosepticum', BMC Genomics, 17, art. #47, pp. 1-15.
