Influences of low temperatures (4 and 8 ° С) on the frost tolerance and fatty acid compositions of cells in a winter wheat suspension culture have been studied. It has been found that treatment of the culture with 4 °C (7 days) did not protect cells from subsequent freezing temperature action (-8 °С, 6 h) and was not accompanied significant changes in the fatty acid composition. On the contrary, the treatment of the culture with the temperature 8 °C (7 days) prevented the death caused by freezing temperature and the content of saturated fatty acids decreased: pentadecanoic acid (by 35,0%), palmitic acid (by 19,9%) and stearic acid (by 65,4%), and the content of α-linolenic acid increased by 94%. That was the cause of the double bond index (DBI) increase by 16%. The role of fatty acids composition changes in the process of increasing frost tolerance in plants are discussed.