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Czajkowski, R.

Publications and source records attributed to Czajkowski, R..

14 recordsLinked to original sources

Beyond kin killing: Dickeya-derived phage-tail-like bacteriocin P2D1 targets phylogenetically distant Pseudomonas spp.

Tailocins, phage-tail-derived bacteriocins, are increasingly recognized as potent mediators of microbial antagonism, yet their ecological scope beyond kin-targeting remains poorly understood. Here, we investigated whether P2D1, a tailocin produced by the plant pathogen Dickeya dadantii 3937, can act against environmental bacteria phylogenetically distant from Dickeya spp. Screening 480 soil and rhizosphere isolates from three distinct plant-associated habitats in Poland, we identified nine Pseudomonas spp. strains susceptible to tailocin P2D1. Whole-genome sequencing and phenotype profiling revealed that these isolates spanned multiple clades, including taxa related to P. germanica, P. tensinigenes, and P. parakoreensis. The D. dadantii mutant lacking genes encoding tailocin sheath and tube proteins lost antagonistic activity against Pseudomonas isolates, confirming that tailocins alone mediate the observed killing. Plant tissue assays revealed that six of the P2D1-susceptible strains were non-pathogenic and could mitigate D. dadantii-induced soft rot on potato. In contrast, three isolates related to P. tensinigenes were able to cause rot on their own under permissive conditions. Together, these findings demonstrate that P2D1 tailocin extends its activity to ecologically co-occurring but taxonomically distant Pseudomonas, suggesting that conserved receptors underline cross-genus targeting. More broadly, our results add to the limited evidence for tailocin activity beyond kin killing and therefore challenge the prevailing paradigm of kin-restricted tailocin specificity. They further suggest that tailocins may influence microbial community assembly across taxonomic boundaries, while their in vivo roles remain understudied.

microbiology↗

Production of P2D1 tailocins by Dickeya dadantii 3937: a temporal relationship between the stressor onset, gene expression, and the concentration of active particles

Tailocins are bacteriocins resembling bacteriophage tails. Previously, we reported the production of P2D1 tailocins in the plant pathogen Dickeya dadantii 3937, the synthesis of which was upregulated upon treatment with mitomycin C. In the current study, we characterized the temporal relationship between the inducer onset, activation of expression of tailocin-related genes, the concentration of tailocin particles, and the titer of viable producer cells. Using a newly established RT-qPCR assay, we found that the expression of P2D1 structural genes peaks two hours after the addition of mitomycin C. Simultaneous measurement of the titer of tailocins showed that the maximum concentration of these particles in the culture supernatant was reached 6 hours post induction and remained stable till the termination of the experiment (18 hours). Progressing accumulation of tailocins between the second and the sixth-hour post mitomycin C treatment correlated with a sharp drop in the count of viable cells (ca. five orders of magnitude). This effect was related to the mechanism by which tailocins are released from the cells, which involves cell lysis. Our study is the first detailed analysis of the timing of events during P2D1 tailocin synthesis in Dickeya spp.

microbiology↗

Tailocin-mediated interactions in Soft Rot Pectobacteriaceae

Bacteria carry phage-derived elements within their genomes, some of which can produce phage-like particles (tailocins) used as weapons to kill kin strains in response to environmental conditions. This study investigates the production and activity of tailocins by plant pathogenic bacteria: Pectobacterium, Dickeya, and Musicola genera, which compete for niche, providing an attractive model to study the ecological role of tailocins. Microscopy revealed that most analyzed strains (88%) produced tailocins. Tailocin-mediated killing interactions were assessed across 351 strain pairs, showing that Dickeya spp. had a higher likelihood of killing neighbors (57.1%) than Pectobacterium spp. (21.6%). Additionally, Dickeya spp. strains exhibited broader phylogenetic killing, targeting both Pectobacterium spp. and Musicola sp., while Pectobacterium spp. tailocins were genus-specific. Mutual killing was observed in 33.9% of interactions, predominantly within Dickeya spp. Although tailocins were morphologically indistinguishable between producers, genomic analyses identified conserved clusters having distinct differences between Pectobacterium spp. and Dickeya spp tailocins. This suggests different origins of these particles. Induction experiments demonstrated that tailocin production was boosted by hydrogen peroxide, supporting the role of these particles in bacteria-bacteria competition during infection. Tailocins were detectable in infected potato tissue but not in river water, highlighting the particular ecological relevance of tailocins in plant environments.

microbiology↗

Inner membrane protein OutB is covalently attached to peptidoglycan in the γ-proteobacterium Dickeya dadantii

Gram-negative bacteria possess a multilayered envelope comprising an inner membrane (IM), a thin peptidoglycan (PG) layer, and an outer membrane (OM). In Escherichia coli and certain other {gamma}-proteobacteria, Braun lipoprotein (Lpp) covalently tethers the OM to PG. Only a few other OM proteins have been found to be covalently linked to PG in Gram-negative bacteria. Here, we showed that in the phytopathogenic {gamma}-proteobacterium Dickeya dadantii, an IM protein, OutB, is covalently attached to PG, thereby tethering itself and the associated type 2 secretion system to the cell wall. In contrast to Lpp, OutB reaches the PG layer from the IM side. By modifying the length of Lpp, which would displace the PG layer in the periplasm, we found that the elongated Lpp+21 improved OutB attachment to PG, whereas the shortened Lpp{Delta}21 reduced it. We showed that two L,D-transpeptidases, Ldt03 and Ldt84, tether Lpp and OutB to PG by the same catalytic mechanism involving the formation of an amide bond between their C-terminal lysine and the stem peptide. Ldt03 and Ldt84, each display substrate specificity for the type of peptide stem and preferentially cross-link Lpp to monomeric and dimeric muropeptide, respectively. The C-terminal Lpp-like box of OutB is almost identical to that of Lpp; it tolerates substantial amino acid substitutions and allows PG-tethering of a bona fide periplasmic protein. Thus, it seems possible that the repertoire of periplasmic and membrane proteins tethered to PG may be more extensive than currently assumed.

microbiology↗

Two complementary circuits for spatial navigation in mouse: evidence from Fos mapping

We designed and implemented a novel spatial behavioral protocol based on the figure 8 maze paradigm. This solution employs a fully automated closed-loop system using visual contexts for guiding behavior. It allows high throughput testing of large cohorts of animals. This automated approach highly reduces the effect of stress, but also eliminates the influence of rule learning and the involvement of working memory. Here, we show that training mice in the apparatus leads to change in strategy from internal tendency to alternate into navigation based exclusively on visual cues. This effect could be achieved using two different protocols: prolonged alternation training, or a flexible protocol with unpredictable turn succession. We provide evidence of opposing levels of engagement of hippocampus and retrosplenial cortex after training mice in these two different regimens. This supports the hypothesis of the existence of parallel circuits supporting navigation, one based on the well described hippocampal representation, and another, RSC-dependent.

neuroscience↗

Soft rot pathogen Dickeya dadantii 3937 produces tailocins resembling the tails of Enterobacteria bacteriophage P2

Tailocins are nanomolecular machines with bactericidal activity. They are produced by bacteria to contribute to fitness in mixed communities, and hence, they play a critical role in their ecology in a variety of habitats. Here, we characterized the new tailocin produced by Dickeya dadantii strain 3937, a well-characterized member of plant pathogenic Soft Rot Pectobacteriaceae (SRP). Tailocins induced in D. dadantii were ca. 166 nm long tubes surrounded by contractive sheaths with baseplates having tail fibers at one end. A 22-kb genomic cluster involved in their synthesis and having high homology to the cluster coding for the tail of the Enterobacteriophage P2 was identified. The D. dadantii tailocins, termed dickeyocin P2D1 (phage P2-like dickeyocin 1), were resistant to inactivation by pH (3.5 - 12), temperature (4 - 50 {degrees}C), and elevated osmolarity (NaCl concentration: 0.01 - 1 M). P2D1 could kill a variety of different Dickeya spp. but not any strain of Pectobacterium spp. tested and were not toxic to Caenorhabditis elegans. TeaserTailocins are nanomolecular entities similar to syringes that are produced by various bacteria to fight other microorganisms present in the same environment.

microbiology↗

Spatio-temporal mechanisms of consolidation, recall and reconsolidation in reward-related memory trace

The formation of memories is a complex, multi-scale phenomenon, especially when it involves integration of information from various brain systems. We have investigated the differences between a novel and consolidated association of spatial cues and amphetamine administration, using an in-situ hybridisation method to track the short-term dynamics during the recall testing. We have found that remote recall group involves smaller, but more consolidated groups of neurons, which is consistent with their specialisation. By employing machine learning analysis, we have shown this pattern is especially pronounced in the VTA; furthermore, we also uncovered significant activity patterns in retrosplenial and prefrontal cortices, as well as in the DG and CA3 subfields of the hippocampus. The behavioural propensity towards the associated localisation appears to be driven by the nucleus accumbens, however, further modulated by a trio of the amygdala, VTA and hippocampus, as the trained association is confronted with test experience. These results show that memory mechanisms must be modelled considering individual differences in motivation, as well as covering dynamics of the process.

neuroscience↗

Being spontaneous has its costs! Characterization of the spontaneous phage ΦD5-resistant mutants of Dickeya solani strain IPO 2222

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 D. solani IPO 2222 mutants (0.8% of all obtained mutants), DsR34 and DsR207, resistant to infection caused by lytic phage vB_Dsol_D5 ({Phi}D5) 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 genes encoding: secretion protein HlyD (mutant DsR34) and elongation factor Tu (EF-Tu) (mutant DsR207) were altered in these strains. Both mutations impacted the proteomes of cells grown in both rich and minimal media, including the abundance of the cell envelope and transmembrane transport-associated proteins. Furthermore, features essential for the ecological success of these mutants in a plant environment, including their ability to use various carbon and nitrogen sources, produce plant cell wall degrading enzymes, ability to form biofilms, siderophore production, swimming and swarming motility and virulence in planta were assessed. Compared to the wild-type strain, D. solani strain IPO 2222, mutants DsR34 and DsR207 had a reduced ability to macerate chicory leaves and to colonize and cause symptoms in growing potato plants. The implications of the {Phi}D5 resistance on driving traits affecting the ecological performance of D. solani are discussed.

microbiology↗

Resistance of Dickeya solani strain IPO 2222 to lytic bacteriophage vB_Dsol_D5 ({Phi}D5) results in fitness tradeoffs for the bacterium during infection

Resistance to bacteriophage infections protects bacteria in phage-full environments, allowing them to survive and multiply in the presence of their viral predators. However, such resistance may cause direct costs for strains linked with the ecological fitness expressed as reduced competitiveness for resources or reduced virulence or both. Unfortunately, limited knowledge exists about such costs paid by phage-resistant plant pathogenic bacteria in their natural environments. This study analyzed the costs of phage resistance paid by broad host phytopathogenic pectinolytic bacterium Dickeya solani both in vitro and in potato (Solanum tuberosum L.) plants. Thirteen D. solani IPO 2222 Tn5 mutants were identified that exhibited resistance to infection caused by lytic bacteriophage vB_Dsol_D5 ({Phi}D5). The genes disrupted in these 13 mutants encoded proteins involved in the synthesis of the bacterial envelope components (viz. LPS, EPS and capsule). The ability of {Phi}D5-resistant D. solani mutants to colonize and cause symptoms on potato plants as well as other phenotypes that are known to contribute to the ecological fitness of D. solani in-plant environment, including growth rate, production of effectors, swimming and swarming motility, use of various carbon and nitrogen sources and biofilm formation were assessed. Although phage resistance did not affect most of the phenotypes of {Phi}D5-resistant D. solani evaluated in vitro, all phage resistant mutants were significantly compromised in their ability to survive on and colonize and cause disease symptoms in potato plants. This study is, to our knowledge, one of few to show the direct link between phage resistance and the fitness of plant pathogenic bacteria and the first one to assess phage-host associations for D. solani.

microbiology↗

High-quality complete genome resource of tomato rhizosphere strain Pseudomonas donghuensis P482, a representative of a species with biocontrol activity against plant pathogens

Strain P482 was isolated from a tomato rhizosphere and classified as Pseudomonas donghuensis. The P. donghuensis species was first established in 2015 and currently comprises only four strains: P482, HYST, SVBP6, and 22G5. P. donghuensis strains antagonize plant pathogens, including bacteria, fungi and oomycetes, and therefore are of high interest regarding their biological control potential to combat plant diseases. The antimicrobial activity of P. donghuensis P482 is based on the production of iron scavenging compound 7-hydroxytropolone, antifungal volatile organic compounds, and yet unidentified secondary metabolite(s). Here, we report a complete genome resource for P. donghuensis strain P482. The genome consists of a single chromosome (5 656 185 bp) with 5258 ORFs (5158 protein-coding genes, 74 tRNAs, 22 rRNAs, 3 ncRNAs and 1 tmRNA) and no plasmid. We believe that the information of the first high-quality, complete genome of P. donghuensis will provide resources for analyses targeting the biological control potential of this species and understanding the traits essential for plant-microbe interaction.

genomics↗

High-quality complete genome resource of plant pathogenic bacterium Pectobacterium atrosepticum strain Green1 isolated from potato (Solanum tuberosum L.) in Greenland

Pectobacterium atrosepticum is a narrow host range pectinolytic plant pathogenic bacterium causing blackleg of potato (Solanum tuberosum L.) worldwide. Till present, several P. atrosepticum genomes have been sequenced and characterized in detail; all of these genomes have come, however, from P. atrosepticum strains isolates from plants grown in temperate zones, not from hosts cultivated under different climatic conditions. Herewith, we present the first complete, high-quality genome of the P. atrosepticum strain Green1 isolated from potato plants grown under subarctic climate in Greenland. The genome of P. atrosepticum strain Green1 consists of one chromosome of 4,959,719 bp., with a GC content of 51% and no plasmids. The genome contains 4531 annotated features, including 4179 protein-coding genes (CDSs), 22 rRNA genes, 70 tRNA genes, 8 ncRNA genes, 2 CRISPRs and 126 pseudogenes. We believe that the information of this first, high-quality, complete, closed genome of P. atrosepticum strain isolated from host plant grown in subarctic agricultural region will provide resources for comparative genomic studies and for analyses targeting climatic adaptation and ecological fitness mechanisms present in P. atrosepticum.

microbiology↗

High-quality complete genome resource of plant pathogenic bacterium Dickeya solani strain IPO 2019 isolated from Hyacinthus orientalis

Dickeya solani is an emerging plant pathogenic bacterium, causing disease symptoms in a variety of agriculturally relevant crop species worldwide. To date a number of D. solani genomes have been sequenced and characterized, the great majority of these genomes have however come from D. solani strains isolated from potato (Solanum tuberosum L.) and not from other plant hosts. Herewith, we present the first complete, high-quality genome of D. solani strain IPO 2019 (LMG 25990) isolated from ornamental plant Hyacinthus orientalis. The genome of D. solani strain IPO 2019 consists of one chromosome of 4,919,542 bp., with a GC content of 56.2% and no plasmids. The genome contains 4502 annotated features, 22 rRNA genes, 73 tRNA genes and 1 CRISPRS. We believe that the information of this high-quality, complete, closed genome of D. solani strain isolated from host plant different than potato (i.e. hyacinth) will provide resources for comparative genomic studies as well as for analyses targeting adaptation and ecological fitness mechanisms present in Dickeya solani species.

microbiology↗

May the phage be with you? Prophage-like elements in the genomes of Soft Rot 3 Pectobacteriaceae (Pectobacterium spp. and Dickeya spp.)

Soft Rot Pectobacteriaceae (SRP; Pectobacterium spp. and Dickeya spp., formerly known as pectinolytic Erwinia spp.) are necrotrophic bacterial pathogens infecting large number of plant species worldwide including agriculturally-important crops. Regardless of the SRP importance in agriculture, little is known about the bacteriophages infecting Pectobacterium spp. and Dickeya spp. and even less about prophages present in SRP genomes. Prophages are recognized as factors underlying bacterial virulence, genomic diversification and ecological fitness and have association with the novel phenotypic properties of bacterial hosts. Likewise, they are recognized as a driving force of bacterial evolution. The aim of this study was to analyze Pectobacterium spp. and Dickeya spp. complete genome sequences downloaded from GenBank (NCBI) for the presence of prophage sequences following their identification and (comparative) characterization with the main focus on current and upcoming perspectives in that field.

microbiology↗

Retrosplenial cortex and its role in spatial cognition

Retrosplenial cortex (RSC) is a region within the posterior neocortical system, heavily interconnected with an array of brain networks, both cortical and subcortical, that is engaged by a myriad of cognitive tasks. Although there is no consensus as to its precise function, evidence from both human and animal studies clearly points to a role in spatial cognition. However, the spatial processing impairments that follow RSC damage are not straightforward to characterise, leading to difficulties in defining the exact nature of its role. In the present article we review this literature and classify the types of ideas that have been put forward into three broad, somewhat overlapping classes: (i) Learning of landmark location, stability and permanence; (ii) Integration between spatial reference frames, and (iii) Consolidation and retrieval of spatial knowledge (\"schemas\"). We evaluate these models and suggest ways to test them, before briefly discussing whether the spatial function may be a subset of a more general function in episodic memory.

animal behavior and cognition↗