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PROVIDER: PRJNA898591 | ENA |
REPOSITORIES: ENA
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Project description:Lytic bacteriophages able to infect and kill Dickeya spp. can be readily isolated from virtually all Dickeya spp.-containing environments, yet little is known about the selective pressure those viruses exert on their hosts. Here, we identified two spontaneous phage-resistant D. solani IPO 2222 mutants, DsR34 and DsR207, resistant to infection caused by phage vB_Dsol_D5 (ΦD5) that expressed a reduced ability to macerate potato tuber tissues compared to the wild-type, phage-susceptible D. solani IPO 2222 strain. Genome sequencing revealed that mutants had point mutations in two genes encoding: secretion protein HlyD (mutant DsR34) and elongation factor Tu (EF-Tu) (mutant DsR207). Both mutations impacted the proteoms of D. solani grown in rich and minimal media. Furthermore, DsR34 and DsR207 were characterized for features essential for their ecological success in a plant environment, including the ability to use various carbon and nitrogen sources, production of plant cell wall degrading enzymes, ability to form biofilms, siderophore production, swimming and swarming motility and virulence in planta. Compared to the wild-type ΦD5-susceptible D. solani strain, mutants DsR34 and DsR207 expressed reduced ability to macerate chicory leaves and to colonize and cause symptoms in growing potato plants. The implications of the ΦD5 resistance on the ecological performance of D. solani are discussed.
2023-04-28 | PXD038825 | Pride
Project description:Pectobacterium are Gram-negative rods of the family Pectobacteriaceae. They are the causative agent of soft rot diseases of crops and ornamental plants. However, their virulence mechanisms are not yet fully elucidated. Membrane vesicles (MVs) are universally released by bacteria and are be-lieved to play an important role in pathogenicity, and survival of bacteria in the environment. Our study investigates the role of MVs in the virulence of Pectobacterium. The results indicate that the morphology and yields of MVs depend on medium composition. In polygalacturonic acid (PGA) supplemented media, Pectobacterium produce MVs of a larger size (100-300 nm) apart of vesicles below 100 nm. Proteomic analyses revealed the presence of pectate degrading enzymes in MVs. The pectate plate test and enzymatic assay proved that those enzymes are active and able to de-grade pectates. What is more, pathogenicity test indicated that MVs derived from Pectobacterium were able to induce maceration of Zantedeschia sp. leaves. We also show that MVs of β-lactamase producing strains were able to suppress ampicillin activity and permit the growth of susceptible bacteria. Those findings indicate that MVs of Pectobacterium play an important role in host-pathogen interactions and niche competition with other bacteria. Our research also sheds some light on the mechanism of MVs production. We demonstrate that Pectobacterium strains, which overexpress the green fluorescence protein (GFP), produce more MVs than wild type strains. Moreover, proteomic analysis revealed that GFP was present in MVs. Therefore, we demonstrate that protein sequestration into MVs is not limited strictly to periplasmic proteins and is a common occurrence. Our research highlights the importance of MVs production as a mechanism of cargo delivery in Pectobacterium and an alternative secretion system.
2022-02-15 | PXD029090 | Pride
Project description:Pectobacterium atrosepticum (Pba) is a gram-negative bacterium which causes blackleg and tuber soft rot on potato. To investigate the molecular processes and responses involved in Pba-host (potato) and Pba-non-host (radish) interactions, under laboratory conditions, we used total RNA-sequencing to measure the gene expression patterns from all three species. Samples from infected and non-infected plant roots were collected after fourteen days of inoculation with Pba SCRI_1039 and subjected to total RNA-sequencing on an Illumina sequencing platform.
2024-01-17 | E-MTAB-13434 | biostudies-arrayexpress
Project description:Bacteriophages of Pectobacterium spp. and Dickeya spp. Genome sequencing
| PRJNA242299 | ENA
Project description:The DsbA oxidoreductase is a crucial factor responsible for introduction of disulfide bonds to the extracytoplasmic proteins in bacteria. A lack of the proper disulfides frequently leads to instability and/or loss of protein function. In pathogens, numerous envelope and extracellular proteins play important roles in pathogenesis; therefore, their improper disulfide bonding may lead to avirulent phenotypes. The importance of the DsbA function in phytopathogens has not been extensively studied yet. Dickeya solani is a bacterium from the Soft Rot Pectobacteriaceae group which is responsible for very high economic losses mainly on potato. In recent years, D. solani became the most abundant potato pathogen among Dickeya species in Europe. In this work, using the D. solani dsbA mutant, we demonstrated that a lack of the DsbA function caused loss of virulence. Mutant bacteria were deficient in most secreted virulence determinants and were not able to develop disease symptoms in the natural host, the potato plant. The SWATH-MS-based proteomic analysis revealed that the dbsA mutation led to multifaceted effects in the D. solani cells. First of all, the levels of the majority of plant cell wall degrading enzymes and proteins related to motility and chemotaxis were severely reduced. Furthermore, the protein profiles suggested induction of the envelope and cytoplasm stress responses in the mutant cells. Finally, the outer membrane barrier seemed to be disturbed by the mutation. Our results clearly demonstrate that the function played by the DsbA oxidoreductase is indispensable for D. solani virulence and a lack of DsbA significantly disturbs cellular physiology. A thorough analysis of proteomic research suggests that a lack of virulence may result from both, abnormalities of the disulfide deprived virulence determinants and the envelope stress-dependent repression of the virulence genes in the dsbA mutant.
2022-02-15 | PXD028047 | Pride
Project description:Importance: Pectobacterium species cause soft rot in potato and other host plants primarily by secreting a battery of plant cell wall degrading enzymes. In addition, several different secretion systems are mobilized during infection. Previous studies of gene expression and regulation thereof primarily focused on the onset of infection. This work investigated transcriptome changes in Pectobacterium during the infection of potato tubers up to 72 hours post inoculation to elucidate biological processes during a longer infection period. Methods: The transcriptomes of aggressive strains of the two species P. carotovorum subsp. carotovorum and P. polaris were investigated during infection of potato minitubers (cv. 'Asterix') at 24, 48 and 72 hours after inoculation by RNA sequencing. The transcriptomes were compared to that of bacteria grown on minimal M9 medium, and transcriptomes from later infection time points (48 and 72 hours after inoculation) were compared to early infection (24 hours after inoculation). Results: Plant cell wall degrading enzymes and secretion system associated genes were largely upregulated during infection compared to in vitro growth, but downregulated in the later phases of infection compared to the early infection phase. The downregulation was not sufficiently explained by the expression of known virulence regulators such as the RsmA/B or the ExpA/S systems.
2022-04-01 | E-MTAB-7921 | biostudies-arrayexpress