One-step genome engineering in bee gut bacterial symbionts

Area of Science:

• Microbiology
• Genetics
• Insect Science

Background:

• Understanding host-microbe interactions in the honey bee microbiome is crucial for bee health.
• Current genome engineering tools for bee gut symbionts are limited.
• Bacterial symbionts play a significant role in honey bee health and crop pollination.

Purpose of the Study:

• To develop a straightforward, single-step genome engineering method for honey bee gut bacteria.
• To enable gene deletions and insertions in the chromosomes of key bee symbionts.
• To facilitate research into host-microbe interactions and develop beneficial symbionts for pollinators.

Main Methods:

• Demonstrated a one-step genome engineering approach using electroporation of DNA with homology regions.
• Successfully performed gene knockouts and insertions in *Snodgrassella alvi* and *Snodgrassella communis*.
• Applied the method to engineer the genome of *Bartonella apis*, an alphaproteobacterium.

Main Results:

• Achieved reliable chromosomal integration for gene deletions and insertions without exogenous recombination machinery.
• Engineered multiple strains of *S. alvi* and two different *Snodgrassella* species.
• Confirmed *recA*-dependence for gene knockouts in *S. alvi*, indicating broad applicability.

Conclusions:

• This lightweight, single-step method simplifies genome engineering in bee gut symbionts.
• The approach is efficient and adaptable to various bee gut bacteria, including *Snodgrassella* and *Bartonella* species.
• Facilitates mechanistic studies of host-microbe interactions and the development of symbionts for pollinator health.