Pectobacterium wasabiae
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PROVIDER: PRJNA206433 | ENA |
REPOSITORIES: ENA
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Project description:Gene expression analysis of wild type Pectobacterium wasabie SCC3193 compared to a SCC3193 inactivation mutant of rsmA
| PRJNA173651 | ENA
Project description:Posttranscriptional regulator RsmA is part of a regulatory system that controls production of plant cell wall degrading enzymes (PCWDEs), motility and other virulence factors among the Pectobacterium spp of plant pathogens. We constructed the knockout mutation of rsmA gene in Pectobacterium wasabiae strain SCC3193. The rsmA- strain has three times lower growth rate but increased virulence when compared to wild type. To explain this phenotype we compared wild type and rsmA- mRNA levels using genome wide microarray. We found that the synthesis of all known virulence factors and flagella is upregulated, while cell division and peptidoglycan synthesis is down regulated in rsmA- strain. The TCA cycle is directed fully to energy production, indicating low energetic status of the cell that is a probable reason for the low growth rate of the mutant. Growth experiments showed that the enzymatic activities of extracellular virulence factors polygalacturonase and pectate lyase are very high throughout the growth curve even without induction by plant component. Wild type cells are rod shaped in liquid media, but elongated and hyperflagellated in swarm plate containing potato tuber extract and in potato tuber. The rsmA- strain exhibits the same conditional phenotype, but differentiation into swarmer cells is faster and cell elongation more pronounced. Virulence experiments in potato tuber showed that although rsmA- strain causes bigger tissue maceration, its ability to compete with the natural bacterial community in the host is decreased. We conclude that the lack of RsmA switches the cells to infective phenotype characterized by expression of virulence factors and swarming. The lack of control over these energy costly processes is probably the reason for decreased growth rate and fitness. Gene expression analysis from P. wasabie SCC3193 wildtype and SCC3193 rsmA mutant was performed on RNA samples extraxcted from the rsmA mutant (at 40 and 48 hours) and wildtype (at 24 hours). The strain were grown on 1.5% agar M9 minimal media plates supplemented with 10% potato extract. Three biological replicates were used from each strain at each time point.
2012-08-24 | E-GEOD-40333 | biostudies-arrayexpress
Project description:Posttranscriptional regulator RsmA is part of a regulatory system that controls production of plant cell wall degrading enzymes (PCWDEs), motility and other virulence factors among the Pectobacterium spp of plant pathogens. We constructed the knockout mutation of rsmA gene in Pectobacterium wasabiae strain SCC3193. The rsmA- strain has three times lower growth rate but increased virulence when compared to wild type. To explain this phenotype we compared wild type and rsmA- mRNA levels using genome wide microarray. We found that the synthesis of all known virulence factors and flagella is upregulated, while cell division and peptidoglycan synthesis is down regulated in rsmA- strain. The TCA cycle is directed fully to energy production, indicating low energetic status of the cell that is a probable reason for the low growth rate of the mutant. Growth experiments showed that the enzymatic activities of extracellular virulence factors polygalacturonase and pectate lyase are very high throughout the growth curve even without induction by plant component. Wild type cells are rod shaped in liquid media, but elongated and hyperflagellated in swarm plate containing potato tuber extract and in potato tuber. The rsmA- strain exhibits the same conditional phenotype, but differentiation into swarmer cells is faster and cell elongation more pronounced. Virulence experiments in potato tuber showed that although rsmA- strain causes bigger tissue maceration, its ability to compete with the natural bacterial community in the host is decreased. We conclude that the lack of RsmA switches the cells to infective phenotype characterized by expression of virulence factors and swarming. The lack of control over these energy costly processes is probably the reason for decreased growth rate and fitness.
2012-08-24 | GSE40333 | GEO
Project description:The phytopathogen Pectobacterium wasabiae SCC3193 is a model for research on virulence regulation in soft rot pathogens. Two proteins acting as global regulators of the expression on genes involved in virulence in SCC3193 are ExpA, a DNA binding transcriptional regulator, and RsmA, an RNA binding post transcriptional regulator. Studies of related enterobacterial phytopathogens suggest that homologues of ExpA (GacA) affect gene expression partly by modulating RsmA-like protein activity. However, the exact extent of this regulon overlap has not been mapped. To elucidate the role of RsmA in ExpA controlled gene expression goes through RsmA in P. wasabiae we performed a gene expression microarray analysis of the wild type strain, an expA mutant, an rsmA mutant, and an expA rsmA double mutant. The microarray-based data of various virulence related genes was confirmed through quantitative RT-PCR. Subsequently, assays were made to link observed transcriptomic differences to changes in growth rate, PCWDE production, and virulence in planta between the four strains. We could conclude that the majority of ExpA regulation seems routed via RsmA modulation. However we could identify genes involved in electron transport, oligogalacturonide transport and metabolism, which ExpA seems to influence independently of RsmA, directly or indirectly. Gene expression analysis from P. wasabie SCC3193 wildtype, SCC3193 rsmA mutant, SCC3193 expA mutant and SCC3193 expA rsmA double mutant was performed on RNA samples extraxcted from the strains at late-logarithmic (late-log) and early stationary (stat) growth phase. Strains were grown in liquid hrp inducing minimal medium cultures supplemented with 0.4% polygalacturonic acid at 200 rpm shaking and 28 degrees celsius. Three biological replicates were used from each strain at each growth phase point.
2013-09-01 | E-GEOD-47545 | biostudies-arrayexpress
Project description:The phytopathogen Pectobacterium wasabiae SCC3193 is a model for research on virulence regulation in soft rot pathogens. Two proteins acting as global regulators of the expression on genes involved in virulence in SCC3193 are ExpA, a DNA binding transcriptional regulator, and RsmA, an RNA binding post transcriptional regulator. Studies of related enterobacterial phytopathogens suggest that homologues of ExpA (GacA) affect gene expression partly by modulating RsmA-like protein activity. However, the exact extent of this regulon overlap has not been mapped. To elucidate the role of RsmA in ExpA controlled gene expression goes through RsmA in P. wasabiae we performed a gene expression microarray analysis of the wild type strain, an expA mutant, an rsmA mutant, and an expA rsmA double mutant. The microarray-based data of various virulence related genes was confirmed through quantitative RT-PCR. Subsequently, assays were made to link observed transcriptomic differences to changes in growth rate, PCWDE production, and virulence in planta between the four strains. We could conclude that the majority of ExpA regulation seems routed via RsmA modulation. However we could identify genes involved in electron transport, oligogalacturonide transport and metabolism, which ExpA seems to influence independently of RsmA, directly or indirectly.
2013-09-01 | GSE47545 | GEO
Project description:Investigation of whole genome gene expression level changes in Aspergillus nidulans OE::rsmA compared to wild-type RDIT9.32 (veA).
2012-01-17 | GSE34220 | GEO
Project description:Pectobacterium wasabiae Genome sequencing
| PRJNA258525 | ENA
Project description:Nontargeted and targeted metabolomics measurements of abiotic stress responses in three-week-old Arabidopsis thaliana plants' rosette leaf tissue for Col-0 wild type plants and double/triple knockout mutants of aquaporins (pip2;1 pip2;2 and pip2;1 pip2;2 pip2;4) treated with drought, heat at different air humidities, or combined drought-heat stress at different air humidities. This experiment contains FT-ICR-MS measurements for 103 Arabidopsis thaliana rosette leaf samples covering three genotypes under six different environmental conditions. The three genotypes comprise the Col-0 wildtype and two loss-of-function mutants of aquaporins, a pip2;1 pip2;2 double mutant and a pip2;1 pip2;2 pip2;4 triple mutant (respective AGI locus identifiers: AT3G53420, AT2G37170, AT5G60660). The six conditions include control condition (well-watered, 22 °C, 70% relative air humidity), drought stress (one week without watering), heat stress without changing the absolute humidity of the ambient air (6 hours at 33 °C, 37% relative air humidity), heat stress with supplemented air humidity to maintain a constant vapor pressure deficit before and during the heat episode (6 hours at 33 °C, 84% relative air humidity), and the combinations of drought pretreatment with each of the two heat stress variants (one week of drought followed by 6 hours of heat stress). Samples from all conditions were harvested at the same time (within 15 min starting at 5 pm). For validation, GC-TOF-MS measurements were done for two genotypes (wildtype, double mutant) and two conditions (drought, control) on partially overlapping samples.
2017-07-17 | MTBLS355 | MetaboLights
Project description:Staphylococcus aureus is a medically important pathogen that exhibit high metabolic versatility allowing it to infect various niches within a host. S. aureus utilizes two major transcriptional regulators, CodY and CcpA, to remodel metabolic and virulence gene expression in response to changing environmental conditions. Previous studies revealed that inactivation of either codY or ccpA has a pronounced impact on different aspects of staphylococcal physiology and pathogenesis. To determine the contribution and interplay of these two regulators in modulating central metabolism, virulence, and biofilm development we constructed and characterized codY ccpA double mutant in S. aureus UAMS-1. In line with previous studies, we found that CcpA and CodY control cellular metabolic status by altering carbon flow through the central and overflow metabolic pathways. Our results demonstrate that ccpA inactivation impairs biofilm formation and decreases incorporation of eDNA into the biofilm matrix while disrupting codY resulted in a robust structured biofilm tethered together with eDNA and PIA. Interestingly, inactivation of both codY and ccpA decreases biofilm biomass and neglects eDNA release in the double mutant. Compared to inactivation of codY, the codY ccpA mutant did not overexpress toxins but maintained overexpression of amino acid metabolism pathways. Furthermore, codY ccpA mutant produced higher amounts of PIA, in contrast to the wild-type strain and ccpA mutant. Overall, results of this study suggest that interplay between CodY and CcpA modulates central metabolism to optimize growth on preferred carbon sources while repressing virulence gene expression until nutrient limitation requires scavenging nutrients from the host.
2022-05-25 | ST002187 | MetabolomicsWorkbench