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Hefeng 25 variety with low vitamin E content in Heilongjiang Province and Bayfield variety with high vitamin E content in Canada were crossed. A total of 144 F2 : 7 recombinant inbred lines (RILs) were used as materials. The genetic linkage mapping of soybean vitamin E was constructed. Soybean varieties were marker-assisted selected in the interval of refined quantitative trait locus (QTLs). QTLs were identified in α-, γ-, δ- and the total tocopherol contents of soybean seeds. Fine QTLs of soybean vitamin E content were identified in the interval between Sat_239 and Satt022 on N linkage group. It was valuable to narrow the interval by marker-assisted selection (MAS). There were seven major QTLs of vitamin E content in soybean. MAS related to vitamin E content in soybean was carried out in the intervals between Sat_239 and Satt022. Considering all the kinds of agronomic traits, six strains with high yield and good quality of vitamin E were chosen, numbered 4, 54, 104, 114, 122 and 135.