The ability of temperature variants of influenza A2 virus to produce plaques in chick embryo kidney tissue culture was studied at different temperatures. Definite differences in efficiency of plaque formation by cryophilic and thermophilic strains were observed at low and high temperatures. Differentiation of the temperature variants appears to reside in a number of genetic markers, designated rct-40, rct-28, and plaque size (S). Virulence of influenza A2 virus, enhanced after prolonged cultivation at high temperature, is probably related to its greater efficiency of plating at 40 C (rct-40(+)), formation of larger plaques at optimal (36 C) and high (40 C) temperatures, and to loss of ability to form plaques at 28 C (rct-28(-)).