Survival of Phytophthora infestans, causal agent of potato and tomato late blight, is thought to be negligible when exposed to freezing conditions typical of a Wisconsin winter. However, the persistence of relatively new P. infestans clonal lineages US-22, US-23, and US-24 within a production region during 2010 to 2014 warranted further investigation. We used tomato seed as a culture medium to determine the survival of P. infestans isolates representing the three lineages under temperatures of 18, 4, 0, -3, and -5°C for 11 time points (1 to 112 days postincubation). Survival varied interactively with temperature, duration of time at a temperature, and clonal lineage of the P. infestans isolate. US-22, -23, and -24 isolates survived for 112 days at 18 and 4°C, 84 days at 0°C, and 14 days at -3°C. US-23 survived longer at -3 and -5°C than did US-22 or US-24. The vigor of US-22 and US-24 isolates decreased with increasing exposure to cold temperatures, a trend that was not observed for the US-23 isolate. By calculating the length of time needed to kill the lineage isolates on infested tomato seed at five temperatures, we predicted that P. infestans would survive in 5% of tomato seed for 99, 25, and 16 days at 0, -3, and -5°C, respectively. We further applied a degree-day model to our empirical data to describe P. infestans survival as a function of cooling degree-day accumulations using archived soil temperatures at 5- and 10-cm depths at four Wisconsin locations over 27 years. The model indicated that survival of P. infestans in 5% of infested tomato seed would occur at 35 and 39% of the location-year combinations at 5- and 10-cm soil depths, respectively. Together, these data suggested that P. infestans has the potential to survive over the winter season by asexual means in infested tomato seed in Wisconsin and other Northern latitudes. Our cooling degree-day model for late blight in the tomato production system offers a tool for anticipating and mitigating disease based on integrated pest management concepts previously utilized for insects.