Discovery and profiling of small RNAs responsive to stress conditions in the plant pathogen Pectobacterium atrosepticum

dc.contributor.authorKwenda, Stanford
dc.contributor.authorGorshkov, Vladimir
dc.contributor.authorRamesh, Aadi Moolam
dc.contributor.authorNaidoo, Sanushka
dc.contributor.authorRubagotti, Enrico
dc.contributor.authorBirch, Paul R. J.
dc.contributor.authorMoleleki, Lucy N.
dc.contributor.emaillucy.moleleki@up.ac.zaen_ZA
dc.date.accessioned2016-05-06T16:02:35Z
dc.date.available2016-05-06T16:02:35Z
dc.date.issued2016-01-12
dc.descriptionAdditional file 1: Table S1. Complete list of RNA-seq detected sRNAs (XLSX 19 kb)en_ZA
dc.descriptionAdditional file 2: Table S2. Predicted transcription start sites of RNA-seq detected sRNAs (XLSX 9114 kb)en_ZA
dc.descriptionAdditional 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)en_ZA
dc.descriptionAdditional 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)en_ZA
dc.descriptionAdditional file 5: Table S5. Conservation analysis in Soft Rot Enterobacteriaceae (XLSX 16 kb)en_ZA
dc.descriptionAdditional file 6: Table S6. Confirmation of RT-PCR amplicons by sequencing and BLASTn against respective sRNA sequences (XLSX 9 kb)en_ZA
dc.descriptionAdditional file 7: Table S7. Differentially expressed sRNA under nutrient-rich and starvation conditions (XLSX 19 kb)en_ZA
dc.descriptionAdditional file 8: Table S8. List of primers used for RT-PCR validation of novel sRNAs (DOCX 12 kb)en_ZA
dc.descriptionAdditional file 9: Table S9. List of primers used for RT-qPCR validation of RNA-seq expression data (DOCX 12 kb)en_ZA
dc.description.abstractBACKGROUND : 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.en_ZA
dc.description.departmentForestry and Agricultural Biotechnology Institute (FABI)en_ZA
dc.description.departmentMicrobiology and Plant Pathologyen_ZA
dc.description.librarianam2016en_ZA
dc.description.sponsorshipThe National Research Foundation (NRF) Thuthuka Grant UID: 69362; UID BFG 93685 and NRF South Africa Russia Bilateral Grant UID: 75252 and from the Russian Foundation for Basic Research (Research Project No. 15-04-02380). The University of Pretoria is acknowledged for providing a studentship for S. Kwenda. V. Gorshkov is supported by The Russian Science Foundation (Project No. 15-14-10022).en_ZA
dc.description.urihttp://www.biomedcentral.com/bmcgenomicsen_ZA
dc.identifier.citationKwenda, 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.en_ZA
dc.identifier.issn1471-2164
dc.identifier.other10.1186/s12864-016-2376-0
dc.identifier.urihttp://hdl.handle.net/2263/52530
dc.language.isoenen_ZA
dc.publisherBioMed Centralen_ZA
dc.rights© 2016 Kwenda et al. Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License.en_ZA
dc.subjectStrand-specific RNA-seqen_ZA
dc.subjectPectobacterium atrosepticumen_ZA
dc.subjectIn silico predictionen_ZA
dc.subjectTranscriptomeen_ZA
dc.subjectRiboswitchesen_ZA
dc.subject5′ UTRen_ZA
dc.subject3′ UTRen_ZA
dc.subjectSmall RNAs (sRNAs)en_ZA
dc.subjectSoft Rot Enterobacteriaceae (SRE)en_ZA
dc.titleDiscovery and profiling of small RNAs responsive to stress conditions in the plant pathogen Pectobacterium atrosepticumen_ZA
dc.typeArticleen_ZA

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