The objective of this study was to evaluate the association of genetic differences in the bovine leptin gene and milk yield, reproduction, body condition score (BCS), and plasma glucose level in Iranian Holstein cows. In total, two hundred and thirty eight cows were used and genotyped for a restricted fragment length polymorphism at the leptin gene locus. Two genotypes, AA and AB, have been distinguished which have the frequencies of 0.89 and 0.11, respectively. The genotypes were distributed according to the Hardy -Weinberg equilibrium (χ 2 = 0.733). During the first 12 wk of lactation, milk yield and composition, live weight, BCS and plasma glucose were measured in 50 cows. Data were analyzed based on a repeated measures ANOVA. During this period, milk yield and composition, live weight, BCS and plasma glucose level were similar among the genotypes. The first cumulative 60-d milk yield, 305-d milk yield, days to first breeding, days open and days from first breeding to conception using previous lactation records were also analyzed using Standard Least Square within mixed models. Fixed effects were year, season, parity and age at calving, and sire. For the reproductive traits the cumulative first 60-d milk yield was also added to the model. Animal was fitted as a random effect. A significant association was detected between the RFLP-AB genotype and 305-d milk yield (p<0.05). The first 60-d cumulative milk yield was similar for the two genotypes (p = 0.21) and tended to be higher in the heterozygous cows. The heterozygous genotypes at the above mentioned locus had a trend to better reproductive performance than the homozygous. The results demonstrate that the RFLP B-allele can yield a higher 305-d milk production with a trend to better reproductive performance.