A total of 96 monoconidial Botrytis cinerea strains were isolated from strawberry field. Physiological characteristics and resistance analysis to boscalid were investigated. Most isolates showed resistant phenotype and the resistance frequency of highly resistant isolates was as high as 45.10％. The effective concentration that inhibited conidia germination by 50％ relative to the control values for sensitive isolates ranged from 0.068 to 60.56 μg ?mL-1 in conidia germination assays. Compared to the sensitive strains, the boscalid-resistant strains had a significant increase in phenylalanine ammonia lyase (PAL) and peroxidase (POD) activity and more sensitivity to boscalid pressure. Fitness analysis showed statistically significant difference between sensitive and resistance isolates. Sequencing of SdhB gene indicated that a single mutation corresponded to His272. The results suggested that resistance to boscalid in B. cinerea populations could result in failure to control gray mold. These results would enrich the understanding of the resistance mechanism of B. cinerea to boscalid.