Abstract: Grain Polyphenol Oxidase (Polyphenol oxidase, PPO) activity is the main factor causing flour and flour products browning. Understanding of allelic variation of PPO gene in bread wheat contributes to improvement of the appearance qualities of the flour products. A total of 118 wheat cultivars from Huanghuai wheat region were used to identify Ppo-A1 alleles with functional marker PPO18 and Ppo-D1 alleles with functional markers PPO16 and PPO29, respectively. The results showed that in the Ppo-A1 locus 48.3% and 51.7% of the wheat varieties possess Ppo-A1a allele (high PPO activity) and Ppo-A1b allele (low PPO activity), respectively, and PPO activities of the Ppo-A1a is significantly higher than that of Ppo-A1b (P＜0.05)．In the Ppo-D1 locus, 55.1% and 44.9% of the wheat varieties possess Ppo-D1a (low PPO activity) and Ppo-D1b (high PPO activity) allele, and PPO activity of the Ppo-D1b is significantly higher than that of Ppo-D1a allele (P＜0.05)．Four types of allelic combinations of Ppo-A1a/Ppo-D1a (intermediate PPO activity), Ppo-A1a/Ppo-D1b (high PPO activity), Ppo-A1b/Ppo-D1a (low PPO activity), Ppo-A1b/Ppo-D1b (intermediate PPO activity) were detected in wheat cultivars surveyed, and their distribution frequencies were 28.8％, 19.5％, 26.3％ and 25.4％, respectively, and the differences among PPO activities of above four genotypes are significant (P＜0.05)．Therefore, the three gene-specific markers are rapid, accurate and convenient to identify allelic variations of PPO gene. This study found some wheat cultivars with low PPO activity, and could provide useful information for improvement of wheat quality.