The invention provides a culture medium composition for saussurea involucrate tissue culture and belongs to the field of biological medicine. The culture medium composition is prepared from a basic culture medium MS, fructose, glucose, agar, 6-BA and 2,4-D. The invention also provides application of the composition. The culture medium composition is mainly used for callus induction or callus subculture of saussurea involucrate, and the culture conditions include temperature of 23-25 DEG C, sunlight time of 9-15 h/d, and light intensity of 1900-2500 lx. The dry weight and total flavonoid content of callus obtained by utilizing the culture medium composition to perform saussurea involucrate tissue culture are remarkably improved compared with the prior art.