Isolation, Culture, and Characterization of Bacteriophages

Pelzek, Adam J.; Schuch, Raymond; Schmitz, Jonathan E.; Fischetti, Vincent A.

doi:10.1002/9780470089941.et0404s07

Cited by 10 publications

(8 citation statements)

References 85 publications

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“…In a manner similar to the classical culture strategies for mammalian viruses in host cells, tissues, and embryonated eggs 30 – 36 , our experiments demonstrate that plant hairy roots can be used for ex vivo propagation of fastidious pathogens like Candidatus Liberibacter spp. We hypothesize similar strategies using insect-vector cell lines could be used for propagating Candidatus Liberibacter spp.…”

Section: Discussionmentioning

confidence: 71%

“…They constitute a valuable tool in plant functional biology, biotechnology, metabolic engineering, molecular pharming, and studies of root-rhizosphere interactions [26][27][28][29] . Classical microbiological techniques developed early in the 19th century, cultured animal and mammalian viruses in host cells, tissues, and embryonated eggs [30][31][32][33][34][35][36] . In a similar manner, we hypothesized that plant hairy roots could be suitable for propagating fastidious pathogens.…”

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confidence: 99%

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Plant hairy roots enable high throughput identification of antimicrobials against Candidatus Liberibacter spp.

Irigoyen¹,

Ramasamy²,

Pant

et al. 2020

Nat Commun

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A major bottleneck in identifying therapies to control citrus greening and other devastating plant diseases caused by fastidious pathogens is our inability to culture the pathogens in defined media or axenic cultures. As such, conventional approaches for antimicrobial evaluation (genetic or chemical) rely on time-consuming, low-throughput and inherently variable whole-plant assays. Here, we report that plant hairy roots support the growth of fastidious pathogens like Candidatus Liberibacter spp., the presumptive causal agents of citrus greening, potato zebra chip and tomato vein greening diseases. Importantly, we leverage the microbial hairy roots for rapid, reproducible efficacy screening of multiple therapies. We identify six antimicrobial peptides, two plant immune regulators and eight chemicals which inhibit Candidatus Liberibacter spp. in plant tissues. The antimicrobials, either singly or in combination, can be used as near- and long-term therapies to control citrus greening, potato zebra chip and tomato vein greening diseases.

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Section: Discussionmentioning

confidence: 71%

mentioning

confidence: 99%

Plant hairy roots enable high throughput identification of antimicrobials against Candidatus Liberibacter spp.

Irigoyen¹,

Ramasamy²,

Pant

et al. 2020

Nat Commun

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“…In the lysogenic cycle, temperate phages integrate their genomes into the bacterial chromosomes, known as lysogenization, as a form of prophages, which can replicate along with the bacterial genome. Temperate phages can also enter the lytic cycle by carefully being cut, or excised, out of the host genome spontaneously or in response to external stresses via induction to release the prophages from the host genome, leading to host lysis [ 8 , 9 ]. The evolutionary purpose of lysogenic phages, their integration into the host genome, and the delayed start of the lytic cycle are multifaceted and, ultimately, serve to produce and spread more infectious phage particles across a bacterial population [ 6 ].…”

Section: Bacteriophages Influence Host Behaviormentioning

confidence: 99%

The Role of Temperate Phages in Bacterial Pathogenicity

et al. 2023

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Bacteriophages are viruses that infect bacteria and archaea and are classified as virulent or temperate phages based on their life cycles. A temperate phage, also known as a lysogenic phage, integrates its genomes into host bacterial chromosomes as a prophage. Previous studies have indicated that temperate phages are beneficial to their susceptible bacterial hosts by introducing additional genes to bacterial chromosomes, creating a mutually beneficial relationship. This article reviewed three primary ways temperate phages contribute to the bacterial pathogenicity of foodborne pathogens, including phage-mediated virulence gene transfer, antibiotic resistance gene mobilization, and biofilm formation. This study provides insights into mechanisms of phage–bacterium interactions in the context of foodborne pathogens and provokes new considerations for further research to avoid the potential of phage-mediated harmful gene transfer in agricultural environments.

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“…1A). Phage collections are readily available (e.g., Felix d’Herelle Reference Center ) or obtainable by traditional isolation procedures (Pelzek et al 2008). Phage resistant, attenuated mutants are then characterized to identify those that fail to adsorb the phage ( i.e.…”

Section: Reviewmentioning

confidence: 99%

Use of bacteriophage to target bacterial surface structures required for virulence: a systematic search for antibiotic alternatives

Orndorff

2016

Curr Genet

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Bacteriophage (phage) that infect pathogenic bacteria often attach to surface receptors that are coincidentally required for virulence. Receptor loss or modification through mutation renders mutants both attenuated and phage resistant. Such attenuated mutants frequently have no apparent laboratory growth defects, but in the host, they fail to exhibit properties needed to produce disease such as mucosal colonization or survival within professional phagocytic cells. The connection between attenuation and phage resistance has been exploited in experimental demonstrations of phage therapy. In such experiments, phage resistant mutants that arise naturally during therapy are inconsequential because of their attenuated status. A more contemporary approach to exploiting this connection involves identifying small effector molecules, identified in high throughput screens, that inhibit one or more of the steps needed to produce a functioning phage receptor. Since such biosynthetic steps are unique to bacteria, inhibitors can be utilized therapeutically, in lieu of antibiotics. Also, since the inhibitor is specific to a particular bacterium or group of bacteria, no off-target resistance is generated in the host’s commensal bacterial population. This brief review covers examples of how mutations that confer phage resistance produce attenuation, and how this coincidental relationship can be exploited in the search for the next generation of therapeutic agents for bacterial diseases.

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Isolation, Culture, and Characterization of Bacteriophages

Cited by 10 publications

References 85 publications

Plant hairy roots enable high throughput identification of antimicrobials against Candidatus Liberibacter spp.

Plant hairy roots enable high throughput identification of antimicrobials against Candidatus Liberibacter spp.

The Role of Temperate Phages in Bacterial Pathogenicity

Use of bacteriophage to target bacterial surface structures required for virulence: a systematic search for antibiotic alternatives

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