Lutein benefits human health significantly, including that of the eyes, skin and heart. Therefore, increasing lutein content in soybean seeds is an important objective for breeding programmes. However, no information about soybean lutein-related quantitative trait loci (QTL) has been reported, as of 2016. The aim of the present study was to identify QTLs underlying the lutein content in soybean seeds. A population including 129 recombinant inbred lines was developed from the cross between ‘Dongnong46’ (lutein 13·10 µg/g) and ‘L-100’ (lutein 23·96 µg/g), which significantly differed in seed lutein contents. This population was grown in ten environments including Harbin in 2012, 2013, 2014 and 2015; Hulan in 2013, 2014 and 2015; and Acheng in 2013, 2014 and 2015. A total of 213 simple sequence repeat markers were used to construct the genetic linkage map, which covered approximately 3623·39 cM, with an average distance of 17·01 cM between markers. In the present study, eight QTLs associated with lutein content were found initially, which could explain 1·01–19·66% of the observed phenotypic variation in ten different tested environments. The phenotypic contribution of qLU-1 (located near BARC-Satt588 on chromosome 9 (Chr 9; linkage group (LG) K)) was >10% across seven tested environments, while qLU-2 (located near Satt192 of Chr 12 (LG H)) and qLU-3 (located near Satt353 of Chr12 (LGH)) could explain 5–10% of the observed phenotypic variation in more than seven environments, respectively. qLU-5, qLU-6, qLU-7 and qLU-8 could be detected in more than four environments. These eight QTLs were novel, and have considerable potential value for marker-assistant selection of higher lutein content in soybean lines.