The rhizosphere is a crucial soil environment where plant-microbe interaction occurs. The increasing use of glyphosate-resistant (GR) soybean may indirectly stimulate detrimental microbial populations and activities in the rhizosphere. Interactions of glyphosate (Gly) on microbial activity were also evaluated. Measurement of rhizosphere microbial activity was based on the urease assay, and C and N mineralization determinations. Gly application on GR soybean may increase phytotoxicity of the herbicide via foliar absorption, thereby decreasing total organic C (TOC) and total organic N (TON) into the rhizosphere. Furthermore, N mineralization varied depending on soil adsorption of Gly and US application to GR soybean.