The Asiatic rice borer Chilo suppressalis (Walker) occurs mainly on rice Oryza sativa L. and water-oat Zizania latifolia (Turcz). Certain ecological and physiological differentiations between rice and water-oat populations have been shown. To determine whether there is host-associated differentiation in supercooling capacity, seasonal changes in supercooling points, glycerol content, and other physiological parameters of naturally occurring C. suppressalis larvae overwintering in rice and water-oat plants were compared over the winter. Supercooling points were low in the winter (November and December), significantly higher in March, and significantly lower in the water-oat population than in the rice population in the winter. Larvae from the water-oat population experienced a significant weight loss from December to March and were heavier than those from the rice population in the winter. Body water content (percentage of fresh weight) was low in the winter and increased significantly in March; no population differences were detected. Hemolymph glycerol content was high in the winter and dropped significantly in March; interpopulation differences were significant in December. Hemolymph trehalose content increased in the winter, and no population differences were detected. Whole body glucose and lipid content varied differently between the rice and the water-oat populations over the winter. It was found that variations in hemolymph glycerol content were responsible for the host-associated seasonal changes in supercooling capacity of overwintering larvae of the Asiatic rice borer.