Rapeseed (Brassica napus L.) is one of most important oilseed crops in the world.There are now various rapeseed cultivars in nature that differ in their seed oil content because they vary in oil-content alleles and there are high-oil alleles among the high-oil rapeseed cultivars.For these experiments, we generated doubled haploid (DH) lines derived from the cross between the specially high-oil cultivar zy036 whose seed oil content is approximately 50% and the specially low-oil cultivar 51070 whose seed oil content is approximately 36%.First, to address the deficiency in polymorphic markers, we designed 5944 pairs of newly developed genome-sourced primers and 443 pairs of newly developed primers related to oil-content genes to complement the 2244 pairs of publicly available primers.Second, we constructed a new DH genetic linkage map using 527 molecular markers, consisting of 181 publicly available markers, 298 newly developed genome-sourced markers and 48 newly developed markers related to oil-content genes.The map contained 19 linkage groups, covering a total length of 2 265.54 cM with an average distance between markers of 4.30 cM.Third, we identified quantitative trait loci (QTL) for seed oil content using field data collected at three sites over 3 years, and found a total of 12 QTL.Of the 12 QTL associated with seed oil content identified, 9 were high-oil QTL which derived from the specially high -oil cultivar zy036.Two high-oil QTL on chromosomes A2 and C9 co-localized in two out of three trials.By QTL mapping for seed oil content, we found four candidate genes for seed oil content related to four gene markers: GSNP39, GSSR161, GIFLP106 and GIFLP046.This information will be useful for cloning functional genes correlated with seed oil content in the future.