A total of 50 bread wheat cultivars or advanced lines from Henan were used to identify allelic variation of vernalization response genes at the Vrn-A1, Vrn-B1, Vrn-D1 and Vrn-B3 loci. The results showed that all wheat cultivars surveyed possessed recessive alleles at the Vrn-B3 locus, and 48 cultivars possessed recessive alleles at the Vrn-A1 and Vrn-B1 loci, and 42 cultivars possessed recessive alleles at the Vrn-D1 locus. It suggests that recessive alleles are the most prevalent at the Vrn-A1, Vrn-B1, Vrn-D1 and Vrn-B3 loci in Henan bread wheat cultivars surveyed. Wheat cultivars with dominant vernalization genes at the Vrn-D1 locus included 8 cultivars of Yunong2019, Yunong 2020, Yunong 2071, Guomai 301, Ping’an 08-8, Bainong 69, Tunmai 3698 and 08 Luo 33. Two new advanced lines Yunong 3052 and Yunong 2053, developed in Henan Agricultural University, showed the deletions of both Vrn-A1 and Vrn-B1 genes. However, these two cultivars still could normally flower in field but they flowered two-days later than Zhoumai 18. It suggests that Vrn-A1 and Vrn-B1 genes are not necessary for bread wheat flowering. This study could provide useful information for screening high-yield and wide-adaptability wheat cultivars in Chinese wheat breeding program.