BACKGROUND: Sorghum (Sorghum bicolor L. Moench) is a cereal crop known for its biological nitrification inhibition (BNI) capacity, a plant-mediated activity limiting nitrification pathway. The use of BNI-producing plants represents an environmentally friendly and cost-effective approach to reduce nitrogen (N) losses, such as nitrate (NO3 -) leaching and nitrous oxide (N2O) gas emissions. The present study aimed to test the effectiveness of different S. bicolor cultivars in rotation to retain ammonium (NH4 +) in soils and promote N availability for the subsequent wheat crop. A two-year field rotation was established with four sorghum cultivars followed by winter wheat (Triticum aestivum L.). Urea alone or combined with the urease inhibitor N-(n-butyl) thiophosphoric triamide was applied to promote a NH4 +-based fertilization regimes.RESULTS: AddingN-(n-butyl) thiophosphoric triamide maintained higher soil NH4 + content and reduced ammonia-oxidizing bacteria population during sorghum cultivation. However, the benefits of the inhibitor on sorghum growth were cultivar-dependent. Notably, the further reduction in ammonia-oxidizing bacteria abundance for sorghum Voyenn and the increased soil NH4 + content for Vilomene suggested a BNI potential for these cultivars. Importantly, the Vilomene precedent enhanced wheat yield for both fertilization regimes.CONCLUSION: Overall, the present study confirms that sorghum is a suitable catch crop and emphasizes the importance of selecting the proper sorghum cultivar to maximize the yield of the target wheat crop, at the same time as minimizing N losses. Furthermore, developing combined strategies with selected sorghum cultivars and the application of urease inhibitors enables to enhance sorghum productivity as forage, achieving added value to the rotation. © 2024 The Author(s). Journal of the Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.