The entire agri-food value chain is facing today a great challenge, which means to move towards a sustainable and more efficient, clean production and trends of consumers under risks and uncertainties of climate change. Good Agricultural Practices (GAP) are necessary to produce fresh vegetables in optimal conditions securing their quality, traceability, and safety. However, the overuses of agrochemicals in the agriculture have increased the levels of contaminants in water bodies generating hazards and risks throughout the agri-food value chain. In Chile, the national surveillance system have reported several notifications of chemical contaminations, due to the presence of pesticide residues exceeding the Maximum Residues Levels (MRLs) and unauthorized pesticides for several crops. Nevertheless, from the perspective of microbial safety, there are no evidences of risks. Therefore, there is a necessity to use sustainable technologies in the agriculture to reduce the presence of pesticide residues in fresh vegetables. In the last decade, the nanotechnology is evolving as a relevant tool in modern agri-food industry, which is significantly advancing to contribute with solutions for production and processing of fresh vegetables. The nanotechnology research fields have generated new agrochemical agents and new delivery mechanisms to improve crop productivity, and it promises to reduce pesticide use. Additionally, novel DNA-based application/devices of nanosensors/nanobiosensors in crop protection have been developed for the plant disease diagnostics and chemical residues. Also, some of these nanobiosensors have been validated with success for traceability and authenticity of fresh vegetables. Novel advances in nanostructures and nanoparticles have demonstrated to be effective in removal of pesticide residues from fresh vegetables, water bodies and soils. Furthermore, nanotechnology is actively contributing in the development of a new generation of intelligent and active packaging together to nanomaterials for storage and transportation of fresh vegetables. This chapter aims to provide a perspective for the further challenges and opportunities that nanotechnology can face to enhance food safety of fresh vegetables in Chile.