Strain variation in competency induction of the core honey bee gut bacterium, Snodgrassella alvi

Many diseases afflict the Western honey bee, Apis mellifera, an important pollinator in agriculture and ecosystems. One way to combat disease is implementation of genes that produce pro-immune response metabolites into bacteria native to the honey bee gut. One core bacterium in the honey bee gut microbial community is Snodgrassella alvi, which has potential to be modified to promote host health, yet research is limited on variability and methods for direct transformation, or uptake of free DNA, by this microbe. Through protocols for taxonomically similar microbes, I will induce competency in several strains of S. alvi, and test for strain variation in transformation efficiency by quantifying colony number compared to the amount of plasmid DNA used, and colony size. To obtain bacterial strains for this study, 46 S. alvi strains were isolated from honey bee guts, and their 16S rRNA genes were sequenced. All strains had >99% 16S rRNA sequence similarity, but may vary in other aspects of their genome, which may influence competency induction. Results will indicate whether selected competency protocols were successful through the growth of transformed S. alvi cells with a kanamycin resistance gene on selective kanamycin plates. Current results show the successful transformation of one strain, S. alvi Y2k_A, with pBBR1MCS-2 vector via heat shock transformation. The development of reliable competency protocols for S. alvi and the selection of the highest transformation efficiency strains will give researchers a necessary tool to further study potential solutions to honey bee disease through symbiont genetic engineering.