Bovine peptidoglycan recognition protein 1 (PGLYRP-1), an important pattern recognition molecule (PRM) of the innate immune system, is an effector molecule in killing different microorganisms directly. To investigate whether the PGLYRP-1 gene was associated with mastitis and milk production traits in dairy cattle, the polymorphism of this gene was analyzed by PCR-RFLP in a population of 524 Chinese Holstein. A total of ten single nucleotide polymorphism (SNP) loci were identified. The association analysis of single SNP locus showed that T-35A, T-12G and G+102C were significantly associated (P<0.05) with somatic cell score (SCS), while G+102C and G+649C were significantly associated (P<0.05) with 305-day milk yield. Association analysis between combined haplotypes and SCS, milk production traits indicated that H3H3 was associated with the lower SCS (P<0.01), and H2H2 was associated with the lower 305-day milk yield (P<0.01). These findings demonstrated that polymorphisms in PGLYRP-1 gene associated with mastitis resistance and 305-day milk yield, and the H3H3 would provide a useful genetic marker of combined haplotypes for mastitis resistance selection and breeding in Chinese Holstein.