Stylosanthes guianensis (Aubl.) Sw. is an economically important pasture and forage legume in tropical regions of the world. Genetic improvement of the crop can be enhanced through marker-assisted breeding. However, neither single nucleotide polymorphism (SNP) markers nor SNP-based genetic linkage map has been previously reported. In this study, a high-quality genetic linkage map of 2572 SNP markers for S. guianensis is generated using amplified-fragment single nucleotide polymorphism and methylation (AFSM) approach. The genetic map has 10 linkage groups (LGs), which spanned 2226.6 cM, with an average genetic distance of 0.87 cM between adjacent markers. Genetic mapping of quantitative trait loci (QTLs) for important agronomic traits such as yield-related and nutritional or quality-related traits was performed using F2 progeny of a cross between a male-sterile female parent TPRC1979 and male parent TPRCR273 with contrasting phenotypes for morphological and physiological traits. A total of 30 QTLs for 8 yield-related traits and 18 QTLs for 4 nutritional or quality-related traits are mapped on the linkage map. Both the high-quality genetic linkage map and the QTL mapping for important agronomic traits described here will provide valuable genetic resources for marker-assisted selection for S. guianensis.