A subtropical insect, Maruca vitrata (F.) (Lepidoptera: Crambidae), is invasive to temperate zones, in which low temperatures during winter would be a serious challenge for colonization. This study assessed cold tolerance and cold-hardening of M. vitrata to understand its overwintering mechanism. Supercooling capacity was confirmed in all developmental stages exhibiting body freezing points at lower than -10°C, in which supercooling points (SCPs) were significantly different among developmental stages, with eggs having the lowest SCP (at -22.5°C). However, all developmental stages suffered significant mortality after being exposed to low temperatures much higher than SCPs. Furthermore, nonfreezing injury increased with elapsed time at 25°C after cold shock. One of the nonfreezing symptoms was a darkening on thorax, which was explained by uncontrolled prophenoloxidase activation. Pre-exposure to 8°C for 1 h significantly increased the survival of both young and old larvae to a low-temperature treatment (-5°C for 1 h). Rapid cold-hardening (RCH) was accompanied by significant increase in hemolymph trehalose concentration. During RCH, trehalose-6-phosphate synthase was significantly upregulated in its expression level.These results suggest that M. vitrata is a freeze-susceptible species and becomes cold-hardy via hypertrehalosemia.