Alfalfa (Medicago sativa) is one of the most important forage crops in the world;however,its molecular genetics and breeding research are hindered due to the lack of a high-quality reference genome.Here,we report a de novo assembled 816-Mb high-quality,chromosome-level haploid genome sequence for ’Zhongmu No.1’alfalfa,a heterozygous autotetraploid.The contig N50 is 3.92 Mb,and 49 165 genes are annotated in the genome.The alfalfa genome is estimated to have diverged from M.truncatula approximately 8 million years ago.Genomic population analysis of 162 alfalfa accessions revealed high genetic diversity,weak population structure,and extensive gene flow from wild to cultivated alfalfa.Genome-wide association studies identified many candidate genes associated with important agronomic traits.Furthermore,we showed that MsFTa2,a Flowering Locus T homolog,whose expression is upregulated in salt-resistant germplasms,may be associated with fall dormancy and salt resistance.Taken together,these genomic resources will facilitate alfalfa genetic research and agronomic improvement.