Roles of metabolites packaged in viral particles on phage replication. (A) One-step growth curve of GVE2 in its host Geobacillus sp. E263. (B) Influence of palmitic amide on GVE2 replication. The GVE2-infected Geobacillus sp. E263 was incubated with palmitic amide which was dissolved in DMSO. DMSO alone was used as a control. At different time post-infection, the GVE2 copy was measured by quantitative real-time PCR. (C) Effects of valine, proline, glutamic acid, threonine, ribose and isobutene glycol on phage replication. The GVE2-infected Geobacillus sp. E263 was added with valine, proline, glutamic acid, threonine, ribose or isobutene glycol dissolved in PBS. PBS alone was used as a control. At different time post-infection, the GVE2 copy was examined. (D) Influence of palmitic amide at different concentrations on GVE2 replication. The GVE2-infected Geobacillus sp. E263 was added with palmitic amide at 10, 100, or 500 μM, followed by the detection of GVE2 copies with quantitative real-time PCR. DMSO was used as a control. In all panels, the statistical significance between treatments were indicated with asterisks (*p < 0.05; **p < 0.01).