Adaptation of Pseudomonas aeruginosa to Phage PaP1 Predation via O-Antigen Polymerase Mutation

Li, Gang; Shen, Mengyu; Yang, Yuhui; Le, Shuai; Li, Ming; Wang, Jing; Zhao, Yan; Tan, Yinling; Hu, Fuquan; Lu, Shuguang

doi:10.3389/fmicb.2018.01170

Cited by 27 publications

(27 citation statements)

References 59 publications

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“…syringae cells for phage φ6 are higher than those obtained by Rombouts et al (2016) for P. syringae pv. porri and phage KIL3 (1.83 × 10 −6 ) or phage KIL4 (3.33 × 10 −6 ) [25], and those obtained by Li et al (2018) for P. aeruginosa strain PA1 and phage PaP1 (3.0 × 10 −5 ) [81].…”

Section: Discussionmentioning

confidence: 91%

Efficiency of Phage φ6 for Biocontrol of Pseudomonas syringae pv. syringae: An in Vitro Preliminary Study

et al. 2019

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Pseudomonas syringae is a plant-associated bacterial species that has been divided into more than 60 pathovars, with the Pseudomonas syringae pv. syringae being the main causative agent of diseases in a wide variety of fruit trees. The most common treatments for biocontrol of P. syringae pv. syringae infections has involved copper derivatives and/or antibiotics. However, these treatments should be avoided due to their high toxicity to the environment and promotion of bacterial resistance. Therefore, it is essential to search for new approaches for controlling P. syringae pv. syringae. Phage therapy can be a useful alternative tool to the conventional treatments to control P. syringae pv. syringae infections in plants. In the present study, the efficacy of bacteriophage (or phage) φ6 (a commercially available phage) was evaluated in the control of P. syringae pv. syringae. As the plants are exposed to the natural variability of physical and chemical parameters, the influence of pH, temperature, solar radiation and UV-B irradiation on phage φ6 viability was also evaluated in order to develop an effective phage therapy protocol. The host range analysis revealed that the phage, besides its host (P. syringae pv. syringae), also infects the Pseudomonas syringae pv. actinidiae CRA-FRU 12.54 and P. syringae pv. actinidiae CRA-FRU 14.10 strains, not infecting strains from the other tested species. Both multiplicities of infection (MOIs) tested, 1 and 100, were effective to inactivate the bacterium, but the MOI 1 (maximum reduction of 3.9 log CFU/mL) was more effective than MOI 100 (maximum reduction of 2.6 log CFU/mL). The viability of phage φ6 was mostly affected by exposure to UV-B irradiation (decrease of 7.3 log PFU/mL after 8 h), exposure to solar radiation (maximum reduction of 2.1 PFU/mL after 6 h), and high temperatures (decrease of 8.5 PFU/mL after 6 days at 37 °C, but a decrease of only 2.0 log PFU/mL after 67 days at 15 °C and 25 °C). The host range, high bacterial control and low rates of development of phage-resistant bacterial clones (1.20 × 10−3) suggest that this phage can be used to control P. syringae pv. syringae infections in plants, but also to control infections by P. syringae pv. actinidiae, the causal agent of bacterial canker of kiwifruit. Although the stability of phage φ6 was affected by UV-B and solar radiation, this can be overcome by the application of phage suspensions at the end of the day or at night.

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Section: Discussionmentioning

confidence: 91%

Efficiency of Phage φ6 for Biocontrol of Pseudomonas syringae pv. syringae: An in Vitro Preliminary Study

et al. 2019

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“…At the same time, K7R R showed virulence decline comparing with K7, which are consistent with the previous views of CPS to serve as primary phage receptor and as well as main virulence factor of K. pneumoniae K7 (Thurow et al, 1975; Paczosa and Mecsas, 2016). Previous studies have shown that rough-type phage-resistant mutant strains are generally accompanied by the loss of polysaccharide synthesis-associated proteins (Le et al, 2014; Li et al, 2018). However, the CPS synthesis-associated proteins that involved in K7R R -like anti-phage mutation are still uncertain.…”

Section: Discussionmentioning

confidence: 99%

“…The LPS or CPS have been recognized as adsorption receptors for many phages (Pickard et al, 2010; Kiljunen et al, 2011; Sorensen et al, 2011). With mutation in LPS or CPS synthetic genes, O antigen or K antigen of phage-resistant strains is generally lost resulting phenotype with comparatively smaller colony and much lower phage adsorption efficiency than the parental wild type strains (Le et al, 2014; Li et al, 2018). Both O antigen and K antigen are involved in phage adsorption on K. pneumoniae (Thurow et al, 1975; Tomás et al, 1987), but which component serves as primary phage adsorption receptor is still inconclusive.…”

Section: Introductionmentioning

confidence: 99%

Three Capsular Polysaccharide Synthesis-Related Glucosyltransferases, GT-1, GT-2 and WcaJ, Are Associated With Virulence and Phage Sensitivity of Klebsiella pneumoniae

Cai

Wang

et al. 2019

Front. Microbiol.

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Klebsiella pneumoniae ( K. pneumoniae ) spp. are important nosocomial and community-acquired opportunistic pathogens, which cause various infections. We observed that K. pneumoniae strain K7 abruptly mutates to rough-type phage-resistant phenotype upon treatment with phage GH-K3. In the present study, the rough-type phage-resistant mutant named K7R R showed much lower virulence than K7. Liquid chromatography-tandem mass spectrometry (LC-MS-MS) analysis indicated that WcaJ and two undefined glycosyltransferases (GTs)- named GT-1, GT-2- were found to be down-regulated drastically in K7R R as compared to K7 strain. GT-1 , GT-2 , and wcaJ are all located in the gene cluster of capsular polysaccharide (CPS). Upon deletion, even of single component, of GT-1 , GT-2 , and wcaJ resulted clearly in significant decline of CPS synthesis with concomitant development of GH-K3 resistance and decline of virulence of K. pneumoniae , indicating that all these three GTs are more likely involved in maintenance of phage sensitivity and bacterial virulence. Additionally, K7R R and GT-deficient strains were found sensitive to endocytosis of macrophages. Mitogen-activated protein kinase (MAPK) signaling pathway of macrophages was significantly activated by K7R R and GT-deficient strains comparing with that of K7. Interestingly, in the presence of macromolecular CPS residues (>250 KD), K7(Δ GT-1 ) and K7(Δ wcaJ ) could still be bounded by GH-K3, though with a modest adsorption efficiency, and showed minor virulence, suggesting that the CPS residues accumulated upon deletion of GT-1 or wcaJ did retain phage binding sites as well maintain mild virulence. In brief, our study defines, for the first time, the potential roles of GT-1, GT-2, and WcaJ in K. pneumoniae in bacterial virulence and generation of rough-type mutation under the pressure of bacteriophage.

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“…The knockout and complementation of galU and wzy were performed as previously described (Le et al, 2014;Li et al, 2018). Briefly, to complement galU in PAO1r-1 and PAO1 galU, the galU gene was PCR amplified using primers galU-C-F and galU-C-R, and the PCR product was digested by BamHI/PstI, and cloned into the BamHI/PstI digested plasmid pUCP24 to generate pucp-galU.…”

Section: Construction Of P Aeruginosa Strainsmentioning

confidence: 99%

Development of a Bacteriophage Cocktail to Constrain the Emergence of Phage-Resistant Pseudomonas aeruginosa

Shen²,

et al. 2020

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With the emergence of multidrug-resistant and extensively drug-resistant bacterial pathogens, phage therapy and other alternative or additional therapeutic modalities are receiving resurgent attention. One of the major obstacles in developing effective phage therapies is the evolution of phage resistance in the bacterial host. When Pseudomonas aeruginosa was infected with a phage that uses O-antigen as receptor, phage resistances typically achieved through changing or loss of O-antigen structure. In this study, we showed that dsRNA phage phiYY uses core lipopolysaccharide as receptor and therefore efficiently kills the O-antigen deletion mutants. Furthermore, by phage training, we obtained PaoP5-m1, a derivative of dsDNA phage PaoP5, which is able to infect mutants with truncated O-antigen. We then generated a cocktail by mixing phiYY and PaoP5-m1 with additional three wide host range P. aeruginosa phages. The phage cocktail was effective against a diverse selection of clinical isolates of P. aeruginosa, and in the short-term constrained the appearance of the phage-resistant mutants that had beleaguered the effectiveness of single phage. Resistance to the 5phage cocktail emerges after several days, and requires mutations in both wzy and migA Thus, this study provides an alternative strategy for designing phage cocktail and phage therapy.

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Adaptation of Pseudomonas aeruginosa to Phage PaP1 Predation via O-Antigen Polymerase Mutation

Cited by 27 publications

References 59 publications

Efficiency of Phage φ6 for Biocontrol of Pseudomonas syringae pv. syringae: An in Vitro Preliminary Study

Efficiency of Phage φ6 for Biocontrol of Pseudomonas syringae pv. syringae: An in Vitro Preliminary Study

Three Capsular Polysaccharide Synthesis-Related Glucosyltransferases, GT-1, GT-2 and WcaJ, Are Associated With Virulence and Phage Sensitivity of Klebsiella pneumoniae

Development of a Bacteriophage Cocktail to Constrain the Emergence of Phage-Resistant Pseudomonas aeruginosa

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