Both sulfur (S) supply and legume-rhizobium symbiosis can significantly contribute to enhancing the efficiency of phytoremediation of heavy metals (HMs). However, the regulatory mechanism determining the performance of legumes at lead (Pb) exposure have not been elucidated. Here, we cultivated black locust ( Robinia pseudoacacia L.), a leguminous woody pioneer species at three S supply levels ( i.e., deficient, moderate, and high S) with rhizobia inoculation and investigated the interaction of these treatments upon Pb exposure. Our results revealed that the root system of Robinia has a strong Pb accumulation and anti-oxidative capacity that protect the leaves from Pb toxicity. Compared with moderate S supply, high S supply significantly increased Pb accumulation in roots by promoting the synthesis of reduced S compounds ( i.e., thiols, phytochelatin), and also strengthened the antioxidant system in leaves. Weakened defense at deficient S supply was indicated by enhanced oxidative damage. Rhizobia inoculation alleviated the oxidative damage of its Robinia host by immobilizing Pb to reduce its absorption by root cells. Together with enhanced Pb chelation in leaves, these mechanisms strengthen Pb detoxification in the Robinia-rhizobia symbiosis. Our results indicate that appropriate S supply can improve the defense of legume-rhizobia symbiosis against HM toxicity.

The prolonged exposure of agricultural soils to heavy metals from wastewater, particularly in areas near industrial facilities, poses a significant threat to the well-being of living organisms. The World Health Organization (WHO) has established standard permissible limits for heavy metals in agricultural soils to mitigate potential health hazards. Nevertheless, some agricultural fields continue to be irrigated with wastewater containing industrial effluents. This study aimed to assess the concentration of lead in soil samples collected from agricultural fields near industrial areas. Subsequently, we determined the lethal concentration (LC50) of lead (Pb) and other heavy metals for two Collembola species, namely Folsomia candida , a standard organism for soil ecotoxicity tests, and comparing it with Proisotoma minuta . The research further examined the toxic effects of lead exposure on these two species, revealing depletion in the energy reservoirs and alterations in the tissue histology of both organisms. The study revealed that lead can induce genotoxic damage as it evidently has moderate binding affinity with the ct-DNA and hence can cause DNA fragmentation and the formation of micronuclei. Elevated lipid peroxidation (LPO) levels and protein carbonylation levels were observed, alongside a reduction in antioxidant enzymes (CAT, SOD & GPx). These findings suggest that lead disrupts the balance between oxidants and the antioxidant enzyme system, impairing defense mechanisms and consequential derogatory damage within microarthropods. The investigation elucidates a complex network of various signaling pathways compromised as a result of lead toxicity. Hence, it presents a novel perspective that underscores the pressing necessity for implementing an integrated risk assessment framework at the investigated site.