TY - JOUR
T1 - Variably lytic infection dynamics of large Bacteroidetes podovirus phi38:1 against two Cellulophaga baltica host strains
AU - Dang, Vinh T.
AU - Howard-Varona, Cristina
AU - Schwenck, Sarah
AU - Sullivan, Matthew B.
N1 - Publisher Copyright:
© 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.
PY - 2015/11/1
Y1 - 2015/11/1
N2 - Bacterial viruses (phages) influence global biogeochemical cycles by modulating bacterial mortality, metabolic output and evolution. However, our understanding of phage infections is limited by few methods and environmentally relevant model systems. Prior work showed that Cellulophaga baltica phage φ{symbol}38:1 infects its original host lytically, and an alternative host either delayed lytically or lysogenically. Here we investigate these infections through traditional and marker-based approaches, and introduce geneELISA for high-throughput examination of phage-host interactions. All methods confirmed the lytic, original host infection (70-80min latent period;approximately eight phages produced per cell), but alternative host assays were more challenging. A 4.5h experiment detected no phage production by plaque assay, whereas phageFISH and geneELISA revealed phage genome replication and a latent period ≥150min. Longer experiments (26h) suggested an 11h latent period and a burst size of 871 by plaque assay, whereas phageFISH identified cell lysis starting at <5h and lasting to 11h, but for only 7% to 21.5% of infected cells, respectively, and with ∼39 phages produced per cell. These findings help resolve the nature of the alternative host infection as delayed lytic and offer solutions to methodological challenges for studying inefficient phage-host interactions.
AB - Bacterial viruses (phages) influence global biogeochemical cycles by modulating bacterial mortality, metabolic output and evolution. However, our understanding of phage infections is limited by few methods and environmentally relevant model systems. Prior work showed that Cellulophaga baltica phage φ{symbol}38:1 infects its original host lytically, and an alternative host either delayed lytically or lysogenically. Here we investigate these infections through traditional and marker-based approaches, and introduce geneELISA for high-throughput examination of phage-host interactions. All methods confirmed the lytic, original host infection (70-80min latent period;approximately eight phages produced per cell), but alternative host assays were more challenging. A 4.5h experiment detected no phage production by plaque assay, whereas phageFISH and geneELISA revealed phage genome replication and a latent period ≥150min. Longer experiments (26h) suggested an 11h latent period and a burst size of 871 by plaque assay, whereas phageFISH identified cell lysis starting at <5h and lasting to 11h, but for only 7% to 21.5% of infected cells, respectively, and with ∼39 phages produced per cell. These findings help resolve the nature of the alternative host infection as delayed lytic and offer solutions to methodological challenges for studying inefficient phage-host interactions.
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U2 - 10.1111/1462-2920.13009
DO - 10.1111/1462-2920.13009
M3 - Article
C2 - 26248067
AN - SCOPUS:84949318191
SN - 1462-2912
VL - 17
SP - 4659
EP - 4671
JO - Environmental Microbiology
JF - Environmental Microbiology
IS - 11
ER -