Through systematic investigation and data collection of Cypripedium plants, we found 15 species of Cypripedium plants in Gansu Province, accounting for 39.47% of Chinese Cypripedium species. Among them, 9 species are endemic to China. The investigation found two new record species of Cypripedium genus in Gansu Province: Cypripedium palangshanense T.Tang et F.T.Wang and Cypripedium plectrochilum Franch.. After sorting out, the key to the species of Cypripedium in Gansu Province was recompiled.