Lead accumulation and concomitant reactive oxygen species (ROS) scavenging in Robinia pseudoacacia are dependent on nitrogen nutrition.

Heavy metal pollution combined with nitrogen (N) limitation is a major factor preventing revegetation of contaminated land. Woody N₂-fixing legumes are a natural choice for phytoremediation. However, the physiological responses of woody legumes to lead (Pb) with low N exposure are currently unknown. In the present study, a common Robinia cultivar from Northeast China, inoculated and non-inoculated with rhizobia, was exposed to -Pb or + Pb at moderate (norN) or low N application (lowN). Our results showed that without inoculation, independent of N application, Pb taken up by the roots was allocated to the shoot and inhibited photosynthesis and biomass production. In non-inoculated Robinia, Pb-mediated oxidative stress resulted in reduced H₂O₂ scavenging as indicated by increased ascorbate peroxidase (APX) activity in the leaves and proline contents in the roots, independent of N application. Combined lowN∗Pb exposure significantly increased malondialdehyde (MDA) contents in roots and leaves and enhanced APX and dehydroascorbate reductase activities in leaves compared to individual Pb exposure. Rhizobia inoculation raised the abundance of nodules and promoted Pb uptake by roots. Under Pb exposure, inoculation with rhizobia reduced MDA contents, increased proline contents in leaves and roots and enhanced activity of nitrate reductase in the leaves, independent of N application. Under Pb exposure, nitrogenase activity of inoculated Robinia under low- and norN application were similar indicating that enhanced of N₂-fixation at lowN was counteracted by Pb exposure. These results show that inoculation of Robinia with rhizobia can alleviate Pb toxicity at combined lowN and Pb exposure by reducing oxidative stress.