Homologous recombination in the genetic transformation model organism Streptococcus pneumoniae is thought to be important in the adaptation and evolution of this pathogen. While competent pneumococci are able to scavenge DNA added to laboratory cultures, large-scale transfers of multiple kb are rare under these conditions. We used whole genome sequencing (WGS) to map transfers in recombinants arising from contact of competent cells with non-competent ‘target’ cells, using strains with known genomes, distinguished by a total of ~16,000 SNPs. Experiments designed to explore the effect of environment on large scale recombination events used saturating purified donor DNA, short-term cell assemblages on Millipore filters, and mature biofilm mixed cultures. WGS of 22 recombinants for each environment mapped all SNPs that were identical between the recombinant and the donor but not the recipient. The mean recombination event size was found to be significantly larger in cell-to-cell contact cultures (4051 bp in filter assemblage and 3938 bp in biofilm co-culture versus 1815 bp with saturating DNA). Up to 5.8% of the genome was transferred, through 20 recombination events, to a single recipient, with the largest single event incorporating 29,971 bp. We also found that some recombination events are clustered, that these clusters are more likely to occur in cell-to-cell contact environments, and that they cause significantly increased linkage of genes as far apart as 60,000 bp. We conclude that pneumococcal evolution through homologous recombination is more likely to occur on a larger scale in environments that permit cell-to-cell contact. Author summary Bacteria shuffle their genes far less often than humans do and genes or traits are more directly linked with the singular bacterial parent cell rather than the two parents that are involved in sexual reproduction. However, bacteria do occasionally have sex in the form of homologous recombination by taking up external DNA and incorporating it into their genomes. This happens far less regularly than sexual reproduction happens in human generations but is a known way that bacteria undergo ‘Horizontal gene transfer’. This means that genes can be acquired without being inherited. In this study we show that this form of horizontal gene transfer is more likely to happen in certain environments over others in Streptococcus pneumoniae . In particular, we show that this is more likely to happen in environments that closely mirror the nasopharynx which is the natural habitat of S . pneumoniae .
【초록키워드】 Evolution, Human, Sequencing, Genome, SNPs, Sex, Laboratory, DNA, Whole genome sequencing, Culture, pathogen, cells, nasopharynx, Horizontal gene transfer, Recombination, Cluster, SNP, Bacteria, genomes, adaptation, WGS, Strains, Donor, Bacterial, Streptococcus pneumoniae, Homologous recombination, recombinant, strain, Contact, sexual reproduction, mixed cultures, singular, gene transfer, traits, transfer, organism, cultures, parent, pneumoniae, whole genome, genetic transformation, recombination event, recombinants, while, pneumococcal, co-culture, Cell, pneumococci, significantly increased, thought, significantly, involved, events, arising, undergo, added, less, occur, purified, conditions, mapped, competent, horizontal gene, Millipore, transferred, 【제목키워드】 Contact, recombination event, facilitate,