The invention provides a tomato gray mold antagonistic strain B-04-glu which is characterized by comprising four steps for the preparation: firstly, a strain of Bacillus cereus for high-efficient antagonistic gray mold is filtered from a soil sample infected by gray mold, and is named as B-04; secondly, bacteria solution which contains the B-04 bacteria is cultured; thirdly, ultrapure water, 1mM Hepes and electric shock buffer solution are used for washing in turn; fourthly, beta-1, 3-dextranase genes are transformed into the B-04 bacteria through electric shock method, and then the antagonistic strain B-04-glu is obtained. The antagonistic strain B-04-glu belongs to biological prevention and cure and does not contain compositions such as pyrilamine, pyrrole, acylamide and so on of chemical pesticide, thereby the antagonistic strain B-04-glu has the advantages of innocuity, no environmental pollution, difficult production of drug resistance, no residue, easy production, long effective period, strong curing ability, good application effect, safety on non-targeted organisms and so on and has wide application prospect.