The potato rot nematode, Ditylenchus destructor, may experience extreme environmental stress caused by freezing temperatures when overwintering in the field. A series of laboratory experiments were conducted in order to clarify overwintering strategies of the nematode. The current study aimed to examine the cold tolerance of this species in various aqueous environments as well as in plant tissues in order to determine its ability to survive 24 h exposure to subzero temperatures. Cold tolerance of adults and fourth-stage juveniles was significantly lower than that of younger juveniles. The lower lethal temperature of adults was –15°C. Although external ice formation affects all life stages, a few second-stage juveniles were able to survive temperatures as low as –30°C. The results expand the current understanding of freezing survival in the potato rot nematode and will lead to better comprehension of its ability to withstand subzero temperature conditions.