The intensive use of pesticide and plastic mulches has considerably enhanced crop growth and yield. Pesticide residues and plastic debris, however, have caused serious environmental problems. This study investigated the effects of the commonly used herbicide glyphosate and micrometre-sized plastic debris, referred as microplastics, on glyphosate decay and soil microbial activities in Chinese loess soil by a microcosm experiment over 30 days incubation. Results showed that glyphosate decay was gradual and followed a single first-order decay kinetics model. In different treatments (with/without microplastic addition), glyphosate showed similar half-lives (32.8 days). The soil content of aminomethylphosphonic acid (AMPA), the main metabolite of glyphosate, steadily increased without reaching plateau and declining phases throughout the experiment. Soil microbial respiration significantly changed throughout the entirety of the experiment, particularly in the treatments with higher microplastic addition. The dynamics of soil β-glucosidase, urease and phosphatase varied, especially in the treatments with high microplastic addition. Particles that were considerably smaller than the initially added microplastic particles were observed after 30 days incubation. This result thus implied that microplastic would hardly affect glyphosate decay but smaller plastic particles accumulated in soils which potentially threaten soil quality would be further concerned especially in the regions with intensive plastic mulching application.