The invention provides mildew-proof fresh-keeping paint and a preparation method and application thereof. The mildew-proof fresh-keeping paint is prepared from the following ingredients in parts by weight: 20 to 60 percent of resin, 30 to 70 percent of solvents, 0.5 to 10 percent of mildew-proof agents and 0.3 to 5 percent of auxiliary agents, wherein the resin is at least one from polyurethane, acrylate copolymers, polyvinyl alcohol, polyvinylidene chloride, modified polyvinylidene chloride, ethylene-vinyl acetate copolymers, polyvinyl acetate copolymers and vinyl acetate-butyl ester coplymers; the solvent is at least one from water and ethyl alcohol. Therefore the resin is dissolved into the solvent; the film forming performance of the resin is regulated through an auxiliary agent; the ingredient mixture ratio is controlled, so that a pore-free coating with the compact surface is formed; therefore the moisture is effectively inhibited from entering or going out of the coating; the effects of maintaining the moisture constancy, keeping fresh and preserving the moisture can be achieved. The intersolubility of the resin and the mildew-proof agents is good, so that the mildew-proof agents are sufficiently dispersed into the resin; the mildew-proof agents can enter the cells through microbial cell membranes; the microbes can be eliminated, or the germination is prevented so as to prevent the mycete growth; the long-time stable fresh keeping effect is achieved.