Nitrate (NO3–N), the main plant/microbial nitrogen source, has a fast turnover in soil driven by species transformation (nitrification/denitrification) and phyto/microbiota assimilation. The technique of diffusive gradients in thin films (DGT) is capable of a robust, low disturbance measurement of NO3–N but has not been implemented due to the absence of a binding layer suitable for deployment in soils. In this study, a new styrene divinylbenzene-based absorbent with amine functional groups (SIR-100-HP) was cast into an agarose gel support. The NO3–N ion selectivity of the SIR-100-HP/agarose binding layer was retained in the presence of high multivalent ion concentrations and was used successfully to acquire in situ NO3–N measurements in bulk soil. The kinetics of binding and the maximum binding capacity were determined. The total capacity of the DGT containing the SIR-100-HP/agarose binding phase was 667 μg of NO3–N. The performance of DGT was not affected by varying pH (3–8) or ionic strength (0–0.018 mol L–1), while anion competition effects at concentrations reflecting those in common agricultural soils were found to be negligible. Complete elution (100% efficiency) of NO3–N from the binding phase was achieved using a solution of 5% NaCl. This technique was validated in three contrasting soils. CDGT measurements were in excellent agreement with pore water NO3–N values. Two-dimensional NO3–N mapping of a profile of flooded rice paddy soil demonstrated the potential of this novel methodology for improved characterization of in situ N speciation for further understanding of bioavailability and biogeochemical processes of NO3–N in soils.