Prochlorococcus cells are the numerically dominant phototrophs in the open ocean. Cyanophages that infect them are thus a notable fraction of the total viral population in the euphotic zone, and, as vehicles of horizontal gene transfer, appear to drive their evolution. Here we examine the propensity of three cyanophages -a podovirus, a siphovirus, and a myovirus -to mispackage host DNA in their capsids while infecting Prochlorococcus , the first step in phage-mediated horizontal gene transfer. We find the mispackaging frequencies are distinctly different among the three phages. Myoviruses mispackage host DNA at low and stable frequencies, while podo-and siphoviruses vary in their mispackaging frequencies by orders of magnitude depending on growth light intensity. We attribute this difference to the concentration of intracellular reactive oxygen species and protein synthesis rates. Based on our findings, we propose a model of mispackaging frequency determined by the imbalance between the production of capsids and the number of phage genome copies during infection.
RESULTS AND DISCUSSIONHost DNA mispackaging frequency in cyanophage capsids. Using six different loci on the Prochlorococcus MED4 chromosome (representative of the entire chromosome, see Table 3), we quantified the level of mispackaging of MED4 DNA in the capsids of the three different cyanophages (Fig. 1A, Table 2) during infection (Fig. 1B, C). The packaging strategy of the podovirus is T7-like [26] , the myovirus' is likely T4-like headful packaging [27] while the siphovirus' is unknown, but 490 bp direct terminal repeats suggest a T7-like packaging mechanism [28] . Host DNA was detected in the capsids of all 3 cyanophages (Fig. 1C). The levels of mispackaging are low compared to those reported for high transducing phages in S. aureus [29] for example, but comparable to levels (~10 PPM) reported for the marine Synechococcus cyanophage S-PM2 [30] .
Corresponding gene
Putative functionPrimer sequences (forward and reverse, 5'-3')