Macromolecular substances that cause pitting and other modifications of growing ice crystals were found to be associated with cyanobacterial mats, eukaryotic algae and mosses from Ross Island and the McMurdo Dry Valleys, Antarctica. Ice-pitting activities were largely retained by dialysis membranes with molecular weight cut-offs of up to 300 kDa. Unlike most aqueous solutes, the ice-active molecules were not excluded from the ice phase during freezing. The ice-pitting activities of each of the samples tested was destroyed by exposure to temperatures between 45 and 65°C, suggesting that they have a protein component. Ice-active substances were not found in cyanobacteria or mosses from temperate climates, but ice-activity was found to be associated with mosses from cold habitats in North America. Although the function of the ice-active substances is not known, their apparent confinement to cold environments suggests that they have a cryoprotective role.