Evolution of Bacterial Interspecies Hybrids with Enlarged Chromosomes

Conjugation driven by a chromosomally integrated F-plasmid (high frequency of recombination strain) can create bacteria with hybrid chromosomes. Previous studies of interspecies hybrids have focused on hybrids in which a region of donor chromosome replaces an orthologous region of recipient chromosome leaving chromosome size unchanged. Very little is known about hybrids with enlarged chromosomes, the mechanisms of their creation, or their subsequent trajectories of adaptative evolution. We addressed this by selecting 11 interspecies hybrids between Escherichia coli and Salmonella Typhimurium in which genome size was enlarged. In three cases, this occurred by the creation of an F'-plasmid while in the remaining eight, it was due to recombination of donor DNA into the recipient chromosome. Chromosome length increased by up to 33% and was associated in most cases with reduced growth fitness. Two hybrids, in which chromosome length was increased by the addition of 0.97 and 1.3 Mb, respectively, were evolved to study genetic pathways of fitness cost amelioration. In each case, relative fitness rapidly approached one and this was associated with large deletions involving recombination between repetitive DNA sequences. The locations of these repetitive sequences played a major role in determining the architecture of the evolved genotypes. Notably, in ten out of ten independent evolution experiments, deletions removed DNA of both species, creating high-fitness strains with hybrid chromosomes. In conclusion, we found that enlargement of a bacterial chromosome by acquisition of diverged orthologous DNA is followed by a period of rapid evolutionary adjustment frequently creating irreversibly hybrid chromosomes.