Abstract: Alpha-amylase enzyme is the important member in physiological metabolism of high plant, especially during seed germination stage. The current study identified 19 amy1 genes from 19 hexaploid wheat (Triticum aestivum L.) accessions. The nucleotide and deduced amino acid sequences of amy1 genes were analyzed in detail, respectively. It showed that two main variation types were characterized due to the base substitution and/or indel mutations in genomic nucleotide sequences and the exon and intron domains were presented different variation degree. This study identified 8 clustered haplotypes from hexaploid wheat accessions through characterization of exon domains. The haplotype 8 was the major variation types and the others were low relative frequency. The haplotype 7 was a special kind of haplotypes among amy1 genes in wheat accession PI243793 because of the variation of amino acid sequences at 155-161 sites. These results would contribute to the understanding in functional aspects and efficient utilization of amy1 genes in hexaploid wheat cultivars.