SUMMARY: Various environmental stresses can induce production of reactive oxygen species (ROS) to turn on signaling for proper responses to those stresses. Plasma membrane (PM)‐localized respiratory burst oxidase homologs (RBOHs), in particular RBOHD, produce ROS via the post‐translational activation upon abiotic and biotic stresses. Although the mechanisms of RBOHD activation upon biotic stress have been elucidated in detail, it remains elusive how salinity stress activates RBOHD. Here, we present evidence that trafficking of PM‐localized RBOHD to endosomes and then its recycling back to the PM is critical for ROS accumulation upon salinity stress. ateca4 plants that were defective in recycling of proteins from endosomes to the PM and clc2‐1 and chc2‐1 plants that were defective in endocytosis showed a defect in salinity stress‐induced ROS production. In addition, ateca4 plants showed a defect in transient accumulation of GFP:RBOHD to the PM at the early stage of salinity stress. By contrast, ateca4 plants showed no defect in the increase in the ROS level and accumulation of RBOHD to the PM upon flg22 treatment as wild‐type plants. Based on these observations, we propose that factors involved in the trafficking machinery such as AtECA4 and clathrin are important players in salt stress‐induced, but not flg22‐induced, ROS accumulation.

A growing literature indicates that untreated wastewater from leaky sewers stands among major sources of pollution to water resources of urban systems. Despite that, the quantification and allocation of sewer exfiltration are often restricted to major pipe areas where inspection data are available. In large-scale urban models, the emission from sewer exfiltration is either neglected (particularly from private sewers) or represented by simplified fixed values, and as such its contribution to the overall urban emission remains questionable. This study proposes an extended model framework which incorporates sewer exfiltration pathway in the catchment model for a better justified pollution control and management of urban systems at a nationwide scale. Nutrient emission from urban areas is quantified by means of the Modelling of Nutrient Emissions in River Systems (MONERIS) model. Exfiltration is estimated for public and private sewers of different age groups in Germany using the verified methods at local to city scales, upscaling techniques, and expert knowledge. Results of this study suggest that the average exfiltration rate is likely to be less than 0.01 L/s per km, corresponding to approximately 1 mm/m/year of wastewater discharge to groundwater. Considering the source and age factors, the highest rate of exfiltration is defined in regions with significant proportions of public sewers older than 40 years. In regions where public sewers are mostly built after 1981, the leakage from private sewers can be up two times higher than such from public sewers. Overall, sewer exfiltration accounts for 9.8% and 17.2% of nitrate and phosphate loads from urban systems emitted to the environment, which increases to 11.2% and 19.5% in the case of no remediation scenario of projected defective sewer increases due to ageing effects. Our results provide a first harmonized quantification of potential leakage losses in urban wastewater systems at the nationwide scale and reveal the importance of rehabilitation planning of ageing sewer pipes in public and private sewer systems. The proposed model framework, which incorporates important factors for urban sewer managers, will allow further targeting the important data need for validating the approach at the regional and local scales in order to support better strategies for the long-term nutrient pollution control of large urban wastewater systems.