Seed filling rate of soybean has been shown to be a dynamic process in different developmental stages affected by both genotype and environment. The objective of the present study was to determine additive, epistatic and quantitative trait loci (QTLs)×environment interaction (QE) effects of the QTL underlying a seed filling rate of soybean. One hundred and forty-three recombinant inbred lines (RILs) derived from the cross of Charleston and Dongnong 594 were used with 2 years of field data (2004 and 2005). Eleven QTLs with significantly unconditional and conditional additive (a) effect and/or additive×environment interaction (ae) effect at different filling stages were identified. Of them six QTLs showed positive a effects and four QTLs had negative a effects on the seed filling rate during seed development. aa and aae effects of 12 pairs of QTLs were identified by unconditional mapping from the initial stage to the final stage. Thirteen pairs of QTLs underlying the seed filling rate with aa and aae effects were identified by conditional mapping. QTLs with aa and aae (additive×additive×environment) effects appeared to vary at different filling stages. Our results demonstrated that the mass filling rate in soybean seed were under genetic and environmental control.