Abuscular mycorrhizal fungi are thought to survive adverse environmental conditions primarily as spores. Extraradical mycelia of two Glomus species were produced in fine mesh pouches which excluded roots but not hyphae. The mycelia in these pouches were exposed to freezing conditions, either in the field or in a controlled-temperature chamber. Bioassay plants were grown directly in the pouches and mycorrhizal colonization was assessed after 1 month. The mycelia remained infective in frozen soil over winter. This survival was not dependent on either the presence of root pieces or on the connection of mycelia to roots. Spores were not an effective inoculum in these bioassays. Overwinter survival of mycelia would enable plants to become incorporated into functional mycorrhizal associations early in spring.