Nanotechnology is a rapidly evolving field characterized by materials in the scale 1 to 100 nm, which have unique physicochemical and biological properties. Despite these benefits of nanomaterials, there are increasing reports of toxicity of NMs plants and animals. Toxicity of NMs on humans and animals is widely studied, while there is a dearth of information regarding the toxicity of nanomaterials in plants and their cellular interactions. Studying Nanotoxicity in plants is vital, as plants are a source of food for all living organisms. Key factors influencing these interactions include size, shape, morphology, and surface-to-volume ratio, which are analyzed using various physicochemical characterization techniques. A few of the conventional methods such as seed germination, root, shoot elongation, determination of chlorophyll content, secondary metabolites, and antioxidant enzyme activities are used to assess the toxicity of NMs on plants. But these are not sufficient for understanding the cellular-level interaction with macromolecules. Recently, several histochemical, microscopical, and analytical methods have been developed for understanding the toxicity of NMs in living systems. These methods can fairly accurately predict the toxicity at a molecular and cellular level, which marks a significant advancement in nanotoxicity assessment. This chapter explores the latest developments in techniques and methods for the assessment of nanotoxicity in plants and their implications in the safety evaluation of food products.