Abstract: Natural antimicrobial peptides (AMPs) are promising candidates for developing a generation of new antimicrobials to meet the challenge of antibiotic-resistant pathogens, and they are extensively applied in medicine, food, agriculture and other fields. However, the rapid screening of AMPs from natural products still has many difficulties and challenges, such as low efficiency and high consumption. This paper firstly introduces the action mechanism of AMPs (including membrane action mechanism and non-membrane action mechanism); then, the highly efficient screening methods of AMP are systematically reviewed, including bacterial adsorption, cell membrane chromatography, phospholipid membrane chromatography, capillary electrophoresis, colorimetry, thin layer chromatography, fluorescence screening, high-throughput sequencing, and mining databases screening. In addition, the development direction of efficient exploration of AMP is also prospected. This paper provides a scientific basis for the discovery of AMPs from complex natural product systems.