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

Area of Science:

• Microbiology
• Molecular Biology
• Genetics

Background:

• Bacteriophages (phages) are crucial for microbial evolution and have therapeutic potential.
• Traditional phage host identification methods are limited in scope and throughput.
• Understanding phage-host interactions is vital for microbiome engineering and antimicrobial development.

Purpose of the Study:

• To develop a high-throughput method for identifying phage-host interactions in complex microbial communities.
• To investigate the host range of phage-plasmid P1 and its variations.
• To explore the molecular controls governing phage host specificity.

Main Methods:

• Integration of a synthetic ribozyme for autonomous barcoding into phage-plasmid P1.
• Targeted 16S ribosomal RNA (rRNA) sequencing to detect barcoded phages in host genomes.
• Application of the method in synthetic and wastewater microbial communities.

Main Results:

• Identified Aeromonadales as a novel host order for phage-plasmid P1.
• Demonstrated P1 transduction into pathogenic bacteria.
• Revealed variations in phage host range influenced by replication origins and tail fibers in wastewater communities.

Conclusions:

• Autonomous barcoding in phages is an effective tool for mapping host interactions in situ.
• The study expands the known host range of phage-plasmid P1 and highlights its potential for targeting pathogens.
• Molecular components like replication origins and tail fibers significantly modulate phage host specificity.