In an agricultural context, the use of conventional low-density polyethylene (LDPE) and biodegradable plastic mulch film has been actively promoted, however, the effects on physical and biochemical soil properties, crop growth dynamics, yield, and nutrient cycling of conventional and biodegradable mulch film use in a temperate climate remain largely undetermined. Here, we conducted a field experiment, exploring the effects of no mulch (control), conventional (LDPE), and biodegradable (PLA/PBAT) plastic mulch film on soil and crop (Zea mays L.) nitrogen (N) partitioning after application of 15N-labelled ammonium-nitrate fertiliser. Further, we also investigated the treatment effects on soil physical and biochemical properties (e.g., microbial diversity, compound-specific microbial 15N incorporation, N dynamics), plant development, as well as monitoring the biotic and abiotic degradation of the plastic mulch films. We found that conventional mulch film increased crop yield by 25 % and 15N uptake by 34 % compared to the control, simultaneously reducing 15N retention by 40 % in the topsoil (0–10 cm), but not affecting microbial N use efficiency and N transformation and incorporation into the protein pool. Biodegradable film application resulted in similar biomass (306 ± 14 g plant−1) to both control (275 ± 14 g plant−1) and conventional mulch (344 ± 20 g plant−1) treatments, but significantly reduced 15N crop uptake by 63 % compared to the conventional mulch film. We ascribe this to the accelerated mechanical breakdown and faster degradation of the biodegradable mulch film during the growing season. These findings suggest that current biodegradable plastic mulch film polymer blends may not be a suitable alternative to conventional mulch film in terms of short-term productivity and N use efficiency in a temperate climate for maize production.