In order to study the problem that grapes are vulnerable to microbial infection and decay during storage, a method based on antagonistic Burkholderia contaminans against postharvest Botrytis cinerea of Rosa vinifera was proposed in this paper. The method tested the resistance induction mechanism of Botrytis cinerea after harvest and determined the fruit decay rate treated by antagonistic Burkholderia contaminans. The results showed that the antagonistic bacterium B-1 had bacteriostatic effect on many common pathogens of fruits and vegetables to a certain extent, and the bacteriostatic range was wide. Among them, the inhibition rate of Fusarium moniliforme was 75.5% and that of Botrytis cinerea was 51.2%. After testing, it can be found that antagonistic bacteria have an inhibitory effect on pathogenic fungi and have an effect on phenylpropane metabolic pathway, reactive oxygen species metabolic pathway, and the activities of other resistance-related enzymes. Through comparison, it can be found that the antagonistic Burkholderia contaminans has a strong antibacterial mechanism against Botrytis cinerea of rose grape after harvest. The fruit treated with antagonistic B Burkholderia B-1 has significantly reduced the decay rate and increased the activity of antibacterial active protein.