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<p>Bacteriophages (phages) facilitate gene transfer and microbial evolution in all ecosystems and have applications as tools for engineering microbiomes and as antimicrobials. Historic efforts to map phage hosts, such as plaque assays, are limited to cultured bacteria, are low throughput, and are hard to apply in microbial communities and environmentally-relevant contexts. To overcome these limitations, we integrate a synthetic ribozyme that stores information about participation in horizontal gene transfer in 16S ribosomal RNA (rRNA) into the phage-plasmid P1, and perform targeted 16S rRNA sequencing following transduction to identify phage-host interactions. Experiments in synthetic and wastewater communities reveal Aeromonadales as a previously unreported P1 host order and show P1 transduction into pathogens. In wastewater, host range varies across phagemids having different origins of replication and phage-derived particles having different tail fibers. This work shows how autonomous barcoding can be used in phages to identify the molecular controls on their host range in microbial communities.</p>

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Cross-order detection of bacteriophage transduction in microbial communities using RNA barcoding

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年3月23日

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  • Zachary W. LaTurner,
  • Matthew J. Dysart,
  • Samuel K. Schwartz,
  • Elizabeth Zeng,
  • James Chappell,
  • Jonathan J. Silberg,
  • Lauren B. Stadler

摘要

Bacteriophages (phages) facilitate gene transfer and microbial evolution in all ecosystems and have applications as tools for engineering microbiomes and as antimicrobials. Historic efforts to map phage hosts, such as plaque assays, are limited to cultured bacteria, are low throughput, and are hard to apply in microbial communities and environmentally-relevant contexts. To overcome these limitations, we integrate a synthetic ribozyme that stores information about participation in horizontal gene transfer in 16S ribosomal RNA (rRNA) into the phage-plasmid P1, and perform targeted 16S rRNA sequencing following transduction to identify phage-host interactions. Experiments in synthetic and wastewater communities reveal Aeromonadales as a previously unreported P1 host order and show P1 transduction into pathogens. In wastewater, host range varies across phagemids having different origins of replication and phage-derived particles having different tail fibers. This work shows how autonomous barcoding can be used in phages to identify the molecular controls on their host range in microbial communities.