A long-period duration of seed dormancy helps to reduce pre-harvest sprouting (PHS) damage in common wheat when a long period of rainfall or high humidity occurs. Identification of genes or loci underlying seed dormancy duration is of high importance for investigating genetic mechanism of PHS tolerance of wheat. In the present study, the germination index (GI) of different mapping populations at 5 days (GI5), 15 days (GI15), and 30 days (GI30) after harvest (DAH) was recorded during the 2011–2012, 2012– 2013, 2013–2014, and 2014–2015 cropping seasons. Field sprouting (FS) values were also determined in 2012–2013 and 2014–2015 cropping seasons. Association analysis was performed in 260 wheat varieties and advanced lines differing in seed dormancy (SD) using 557 SSR and 14 gene-specific markers for PHS/SDrelated genes. The result indicated that a total of 47 loci were significantly (P < 0.01) associated with SD using both the general linear model (GLM) and mixed linear model (MLM). Five major loci were consistently detected at different post-ripening stages after harvest, including four for GI5 and GI15 on chromosomes 3AS, 3BL, 5AL, and 5BL responsible for middleduration SD, and one for GI5, GI15, and GI30 on 2AL associated with longer-duration SD. The major locus on 2AL (designated Qsd.ahau-2AL) is likely to be a novel QTL, which was linked to Xwmc658 by mapping analysis in 165 recombinant inbred lines (RILs) derived from the Jing 411 × Wanxianbaimaizi cross and 728 F2 plants from Jimai 20 × Suiningtuotuomai. Furthermore, a CAPS marker CAPS-2AL was developed for Qsd.ahau-2AL locus and was validated using the 728 F2 plants and 201 accessions of the Chinese mini-core collection. Our results also revealed that the TaMFT-like gene on 3AS had a more significant association with dormancy duration than the other six genes underlying SD/PHS resistance in Chinese wheat germplasm. This study provides useful information for marker-assisted selection in wheat breeding and enhances our understanding of molecular genetics of SD in wheat.