Axenic cultures of lichen mycobionts and photobionts were assayed for biological ice nucleation activity (INA), a characteristic which has recently been discovered in some species of lichens collected from nature. Of 14 species of mycobionts, five showed nuclei active at −5°C or warmer, while none of the 13 photobionts tested had INA at that temperature. It appears that biological ice nuclei in lichens are produced primarily by the mycobiont. Rhizoplaca chrysoleuca had the greatest INA. Other lichen fungi showing warm temperature INA were Cladonia boryi, Cladonia rangiferina, Lecanora dispersa, and Pertusaria flavicans. Lichen INA may be involved in moisture uptake and/or frost protection.