Soil selenium (Se) speciation characteristics and their influence on the Se enrichment pattern and physiological characteristics of oilseed rape are poorly understood. We investigated dynamic changes in rhizosphere soil physicochemical properties, Se uptake and partitioning, biomass, and physiological indices in oilseed rape under five exogenous Se condition levels (0, 1.0, 2.5, 5.0, and 10.0 mg kg−1 Se in sodium selenite) using soil cultivation experiments. The rhizosphere pH and dissolved organic carbon in the soil solution were higher than those of the non-rhizosphere soil solution. The total Se, water-soluble Se, exchangeable Se, and organic Se contents in soils, as well as rapeseed root/leaf Se contents, significantly increased with increasing exogenous Se. Under 2.5 mg kg−1 Se, the biomass of rapeseed roots and leaves increased at the sixth week (82% and 58%) and eighth week (48% and 32%), respectively, reaching the highest level. Applications of 5.0 mg kg−1 Se at 6 and 8 weeks significantly increased the glutathione peroxidase activity (49%/82%), and decreased malondialdehyde content (23%/39%). Canonical correlation and ridge regression analyses showed that Se in the rapeseed roots/leaves significantly and positively correlated with water-soluble Se, exchangeable Se, and organic Se in rhizosphere soil. Overall, moderate-concentration Se soil application benefited oilseed rape growth (optimum = 2.5 mg kg−1 Se). Our findings reveal the response of oilseed rape to soil Se application based on plant growth and physiological traits, rhizospheric soil solution properties, and Se speciation transformation.