Biochar-derived persistent free radicals and reactive oxygen species reduce the potential of biochar to mitigate soil N2O emissions by inhibiting nosZ

Biochar amendment has been proven to generally reduce soil nitrous oxide (N2O) emissions. However, the effects and underlying mechanisms of biochar-derived reactive oxygen species (ROS) on soil N2O emissions are still unclear. Thus, we firstly weakened the intensities of persistent free radicals (PFRs) within biochar using triethanolamine (TEA) as a quencher, and then used soil incubation methods to compare the potentials of TEA-quenched and un-quenched biochar in mitigating soil N2O emissions. The TEA-quenched biochar generated less hydrogen peroxide (H2O2) and hydroxyl radical (•OH), while having a higher soil N2O emission mitigation potential, than the un-quenched biochar. The N2O emissions and the N2O/(N2O + N2) ratio were significantly, positively, correlated with the generated H2O2 and •OH contents. These results demonstrated that biochar-derived ROS weakened biochar's mitigation of soil N2O emissions. The specific mechanisms of biochar-derived ROS on soil N2O emissions were further explored by a ROS scavenging experiment. It was found that scavenging H2O2 by catalase efficiently hindered the generation of •OH, resulting in decreases in N2O emissions and the N2O/(N2O + N2) ratio. Meanwhile, biochar-derived ROS exhibited a more severe inhibition on N2O reductase gene (nosZ) expression than that on the expression of genes responsible for N2O production (nirK and nirS), indicating that biochar-derived ROS weakened biochar's mitigation of soil N2O emissions by inhibiting microbial N2O reduction. Our results imply that controlling the content of biochar-derived ROS is a promising strategy to maximize biochar's potential for mitigating soil N2O emission.