Maize root exudate composition alters rhizosphere bacterial community to control hotspots of hydrolase activity in response to nitrogen supply

Improving nitrogen (N) acquisition by crops from soil is essential to reduce fertilization rates whilst maintaining yields. Plants can adapt their nutrient acquisition strategies according to N availability, which also affects soil microbial community structure, functions and activities and relies on the supply of carbon (C) for energy. We hypothesized that N deprivation would create hotspots of N- and C-acquiring hydrolase activities in maize rhizosphere through the effects of altered root exudation on the rhizosphere bacterial community. We grew maize under three N fertilization rates and combined soil zymography with the identification of rhizosphere microbial communities and non-targeted metabolic profiling of root exudates to explore enzyme hotspot formation. The rhizosphere extents of β-1,4-glucosidase (BG) and β-N-acetylglucosaminidase (NAG) activities decreased after N fertilization, narrowing by 48% and 39%, respectively, under typical field N application rates compared to zero application. Rhizosphere extents of enzyme activities were more sensitive to altered N supply than changes in the rates of enzyme activities: BG activity decreased by ∼10%, while NAG activity was unaffected. Decreases in the activities of both hydrolases and their rhizosphere extents caused by N addition correlated with reduced abundances of oligotrophs. The relative abundances of oligotrophic bacteria (e.g., Acidobacteria) decreased, while copiotrophs (e.g., Pseudomonadota and Patescibacteria) increased under the highest N application rate. Co-occurrence networks of the rhizosphere bacterial community revealed that functional units increased with BG activity, while an efficient and denser co-occurrence network supported expansion of its rhizosphere extent. The metabolic profiles of root exudates changed according to the N application rate, suggesting that their chemistry was regulated by the plant in response to N supply. The composition of root exudates and dissolved organic C and nitrate contents explained the largest variations in NAG hotspots in the rhizosphere. In summary, maize actively adjusts the composition of root exudates to increase interactions with rhizosphere bacteria, thereby stimulating hydrolase production and activities, and altering their rhizosphere extents to mobilize N and energy (C) in a larger soil volume, under conditions of N deficiency.