The invention discloses a preparation method of a trametes versicolor fermentation extract and application thereof to a printing and dyeing antifoaming agent. The preparation method of the extract comprises the following steps of centrifuging trametes versicolor fermentation broth, filtering supernatant liquor, then filtering by using a ceramic membrane, carrying out discoloration treatment by adopting macroporous resin after rotary-evaporation concentration, transferring a rotary-evaporation concentrated solution subjected to the discoloration treatment to a sephadex column, carrying out elution by using deionized water as eluant, then carrying out resin adsorption on the eluant by adopting a two-effect series process, carrying out desorption by using an ethanol aqueous solution, collecting a desorbed fraction, thus obtaining a concentrated solution through concentration, and finally carrying out vacuum drying or spray drying on the concentrated solution, so that a trametes versicolorfermentation broth extract is obtained. The prepared trametes versicolor fermentation broth extract is applied to the printing and dyeing antifoaming agent, the quick antifoaming performance and thedurability of the antifoaming agent can be improved; and the service performance of the printing and dyeing antifoaming agent is improved.