The study was conducted to evaluate in vitro pollen germination of 50 spring wheat (Triticum aestivum L.) accessions at three storage temperatures (23 °C, 5 °C, and -20 °C). Germination of the mature pollen was measured right after harvest (0 time), and 24, 48, and 72 h after storage (HAS). Differences in fresh pollen germination between accessions were significant and ranged from 21.15% to 1.09%. Pollen germination was the lowest at 24 h at room temperature. The pollens of 24 wheat accessions (AAC Scotia, AW636, AW725, AW780, AW804, AW822, AW823, Brookfield, BRS Pardela, ECSW05, ECSW38, ECSW48, ECSW49, ECSW69, Hoffman, Kleos, Major, Nass, 12NQW-237, 12NQW-413, 12NQW-414, 12NQW-436, 12NQW-754, and 12BW0374) were viable after 24 h when they were stored at 5 °C. Sub-zero temperature did not prolong pollen viability.