Highlights: Land use is the main factor explaining N cycle genes abundance and GHG fluxes. Soil aggregates size is a minor factor explaining N genes abundance and GHG fluxes. Cropland showed the lowest abundance for bacteria, fungi, nifH, narG, nirS and nosZ. Effect of aggregate sizes on N genes abundance was only found in forest sites. Aggregates 0.5–1.0 mm showed the highest N functional genes abundance in forest sites. Soil structure is known to influence microbial communities in soil and soil aggregates are the fundamental ecological unit of organisation that support soil functions. However, still little is known about the distribution of microbial communities and functions between soil aggregate size fractions in relation to land use. Thus, the objective of this study was to determine the gene abundance of microbial communities related to the nitrogen cycle and potential greenhouse gas (GHG) fluxes in six soil aggregate sizes (0–0.25, 0.25–0.5, 0.5–1.0, 1–2, 2–5, 5–10 mm) in four land uses (i.e. grassland, cropland, forest, young forest). Quantitative-PCR (Q-PCR) was used to investigate the abundance of bacteria, archaea and fungi, and functional guilds involved in N-fixation (nifH gene), nitrification (bacterial and archaeal amoA genes) and denitrification (narG, nirS, and nosZ genes). Land use leads to significantly different abundances for all genes analysed, with the cropland site showing the lowest abundance for all genes except amoA bacteria and archaea. In contrast, not a single land use consistently showed the highest gene abundance for all the genes investigated. Variation in gene abundance between aggregate size classes was also found, but the patterns were gene specific and without common trends across land uses. However, aggregates within the size class of 0.5–1.0 mm showed high bacterial 16S, nifH, amoA bacteria, narG, nirS and nosZ gene abundance for the two forest sites but not for fungal ITS and archaeal 16S. Keywords: Quantitative-PCR ; Nitrogen-fixation ; Nitrification ; Denitrification ; Soil aggregates ; Land use