Abstract: Trehalose is a non-reducing disaccharide that has a variety of biological functions in insects or fungi such as participating in energy metabolism, recovery from stress, and chitin synthesis. Trehalase (EC 3.2.1.28) has become a safe candidate target for the development of novel agrochemicals due to its important roles in regulating the metabolism and the content of trehalose, as well as its functional differences between pests and mammals. In this review, the crystal structures of trehalase and its interaction mechanism with inhibitors are summarized. Meanwhile, the research progress of trehalase inhibitors in agriculture is overviewed including validamycin, natural products salbostatin and trehazolin, deoxynojirimycin, natural alkaloids, piperine, and their corresponding synthetic analogues. In addition, the application of piperine-based compounds in the control of plant diseases or pests is highlighted. This review would provide a reference for the design and the discovery of novel piperine-based agrochemicals targeting the structure of trehalase.