Chrysanthemum (Chrysanthemum morifolium Ramat.), one of the four most popular cut flowers in the world, occupies a large share of global floriculture trade. Leaf angle (LA) is an important factor affecting plant architecture, photosynthesis and yields of cut chrysanthemum. However, the genetic basis and related genes underlying LA remain largely unknown. In this research, phenotypes of five LA-related traits in 200 cut chrysanthemum accessions were investigated across two consecutive years (Y2020 and Y2021), and their broad-sense genetic heritabilities ranged from 68.91% to 84.70%. Leaf orientation value (LOV) exhibited the maximum phenotypic coefficient of variation of 35.66% in Y2020 and 32.34% in Y2021. A new methodological framework named 3VmrMLM was applied for multi-locus genome-wide association study (GWAS) based on a set of 330,710 single nucleotide polymorphisms (SNPs) to explore the genetic loci and candidate genes of LA. As a result, 303 quantitative trait nucleotides (QTNs) in single-environment, 120 QTNs and 75 QTN × environments interactions (QEIs) in multiple-environments were identified. Among the genes around these QTNs and QEIs, 26 known genes were considered to be associated with LA, including ARF16, IBH1, AN3, etc. Furthermore, the haplotype analysis of ARF16 revealed four accessions with elite LA. Our findings provide a fundamental understanding to the genetic architecture of LA and contribute to further gene cloning and molecular improvement of chrysanthemum plant architecture.