The invention relates to the technical field of insect-resist agents. The invention relates to an insect repellent, in particular to a biological insect repellent and a preparation method and application thereof. The biological insect-resist agent is prepared from the following raw materials: cortex meliae, star anise, dry red peppers and fermentation strains; the fermentation strains comprise saccharomycetes, lactic acid bacteria, photosynthetic bacteria and actinomycetes. 100 kg of fermentation liquor needs following raw materials: 0.5-0.8 kg of cortex meliae, 0.5-0.8 kg of star anise, 0.5-0.8 kg of dry red peppers and 5-6 kg of EM.1 strains; according to the biological insect-resist agent, the raw materials are simple and easy to obtain, and after the biological insect-resist agent is sprayed to leaf surfaces, metabolism of plants can be enhanced, protective film cuticles of the leaves are strengthened, and pathogenic bacteria are prevented from entering the leaves; in addition, theleavening agent contains an ester component, which is not decomposed in the body of the herbivorous insect and generates physiological barrier damage and is lethal; therefore, due to the action of stimulating substances such as capsaicine, the insect can feel uncomfortable and escape from the body, the insect preventing and expelling effect is obvious, and the preparation process is simple, convenient to use, low in production cost and suitable for industrial development.