Nitrogen Addition Decreases Dissimilatory Nitrate Reduction to Ammonium in Rice Paddies

Abstract

Dissimilatory nitrate reduction to ammonium (DNRA), denitrification, anaerobic ammonium oxidation (anammox), and biological N₂ fixation (BNF) can influence the nitrogen (N) use efficiency of rice production. While the effect of N application on BNF is known, little is known about its effect on NO₃^- partitioning between DNRA, denitrification, and anammox. Here, we investigated the effect of N application on DNRA, denitrification, anammox, and BNF and on the abundance of relevant genes in three paddy soils in Australia. Rice was grown in a glasshouse with N fertilizer (150 kg N ha^-1) and without N fertilizer for 75 days, and the rhizosphere and bulk soils were collected separately for laboratory incubation and quantitative PCR analysis. Nitrogen application reduced DNRA rates by >16% in all the soils regardless of the rhizospheric zone, but it did not affect the nrfA gene abundance. Without N, the amount and proportion of NO₃^- reduced by DNRA (0.42 to 0.52 μg g^-1 soil day^-1 and 45 to 55%, respectively) were similar to or higher than the amount and proportion reduced by denitrification. However, with N the amount of NO₃^- reduced by DNRA (0.32 to 0.40 μg g^-1 soil day^-1) was 40 to 50% lower than the amount of NO₃^- reduced by denitrification. Denitrification loss increased by >20% with N addition and was affected by the rhizospheric zones. Nitrogen loss was minimal through anammox, while BNF added 0.02 to 0.25 μg N g^-1 soil day^-1 We found that DNRA plays a significant positive role in paddy soil N retention, as it accounts for up to 55% of the total NO₃^- reduction, but this is reduced by N application.IMPORTANCE This study provides evidence that nitrogen addition reduces nitrogen retention through DNRA and increases nitrogen loss via denitrification in a paddy soil ecosystem. DNRA is one of the major NO₃^- reduction processes, and it can outcompete denitrification in NO₃^- consumption when rice paddies are low in nitrogen. A significant level of DNRA activity in paddy soils indicates that DNRA plays an important role in retaining nitrogen by reducing NO₃^- availability for denitrification and leaching. Our study shows that by reducing N addition to rice paddies, there is a positive effect from reduced nitrogen loss but, more importantly, from the conversion of NO₃^- to NH₄⁺, which is the favored form of mineral nitrogen for plant uptake.

Keywords: denitrification; dissimilatory nitrate reduction to ammonium; nitrogen; nrfA; rice paddies.

FIG 1

NH₄⁺ (A) and NO₃⁻ (B) concentrations in rhizosphere soil (rh) and bulk soil (nrh) from the three sites (Finley, Jerilderie, and Coree) with (+N) and without (−N) N input. The results for bars with different letters on top are significantly different (P < 0.05). Error bars represent ±1 standard error.

FIG 2

Rate of N transformation by DNRA (A), denitrification (B), BNF (C), and anammox (D) in the paddy soils from the three sites (Finley, Jerilderie, Coree). +N and −N, with and without N input, respectively; rh and nrh, rhizosphere and bulk soils, respectively. The result for bars with different letters on top within a chart are significantly different (P < 0.05). Error bars represent ±1 standard error. The y axis scales are different for the different panels.

FIG 3

Abundance of the nrfA (A), nosZ (B), nifH (C), and hzsB (D) genes in the three sites (Finley, Jerilderie, and Coree). +N and −N, with and without N input, respectively; rh and nrh, rhizosphere and bulk soils, respectively. The results for bars with different letters on top within each panel are significantly different (P < 0.05). Error bars represent ±1 standard error.

FIG 4

Relationship between the C/NO₃⁻ ratio and N transformation rates (A) and the proportion of NO₃⁻ reduced by DNRA (B).