Shoot-fresh-weight (SFW) is one of the parameters, used to estimate the total plant biomass yield in soybean. Understanding the genetic and molecular basis of SFW could help increase the total biomass production. In this particular study, we identified QTLs associated with SFW in a Recombinant Inbred Line (RIL) population derived from interspecific cross of PI483463 and Hutcheson. A total of 551 (535 SNP and 16 SSR) markers, were found to be polymorphic between the parental lines and were used to screen the RILs to develop the genetic map. Linkage analysis and QTL mapping were performed using with the software QTL IciMapping version 4.0, with the minimum LOD score of 3.0 and estimating the likelihood of a QTL and its corresponding effects at every 1cM. QTLs with LOD value > threshold LOD, as determined by 1000 permutation tests at p > 0.05 were considered as significant QTLs. The analysis identified a total of 5 QTLs associated with shoot fresh weight over two environments, with the phenotypic variation (PV) ranging from 6.34 to 21.32%, and the additive effect from -0.54 to 0.33. Among these QTLs, qFW1314_19_1 had the largest LOD scores, with PV of 21.32%. Interestingly, three QTLs, qFW2013_19_1, qFW2014_19_1, and qFW1314_19_1 identified on chromosome 19(L), showed negative additive effects, indicating the contribution from the wild parent PI483463. The QTLs identified in this study can be the targets to identify the candidate genes for the SFW and can help in developing cultivars with increased biomass potential.