V. natriegens cells before low-temperature preservation
WT V. natriegens was first streaked onto LB3 agar to obtain single colonies. Subsequently, a single colony was inoculated into LB3 liquid medium and incubated in an orbital incubator at 30C for 12 h. Aliquots of the culture were then inoculated into three independent conical flasks containing 100 mL LB3 liquid media at a ratio of 1:200 (v/v). After 4 h of incubation, the cultures reached the late exponential phase. A portion of the cells were withdrawn for total RNA extraction (CK), while the rest of the cultures were preserved at 4C. Three biological replicates were used for the transcriptomic analysis.