Haplotype-resolved chromosome-level genome assembly of creeping bentgrass, Agrostis stolonifera

Creeping bentgrass (Agrostis stolonifera) is a widely used cool-season turfgrass valued for its fine texture and ability to form dense, uniform turfs. However, its complex allotetraploid genome and high repetitive content have posed challenges for genomic research and molecular breeding. Here, we report a haplotype-resolved chromosome-level genome assembly generated using PacBio HiFi and Oxford Nanopore sequencing with Omni-C scaffolding. The final assembly spans 5.4 Gb, with a scaffold N50 of 187.9 Mb and comprises 28 pseudochromosomes representing fully phased haplotypes (2n = 4x = 28). BUSCO analysis indicated 98.8% completeness, indicating the high quality of the assembly. We annotated 146,216 protein-coding genes and found that transposable elements account for 79.8% of the genome, dominated by LTR-Gypsy elements. Subgenome-specific LTR clustering and comparative genomic alignments supported an allopolyploid origin involving two diverged progenitors. This high-quality genome provides a foundational resource for functional genomics and breeding efforts to improve disease resistance, abiotic stress tolerance, and turf quality.