Abstract: Tenuazonic acid(TeA) can be used to control aphid, which has obvious effect. In order to further develop and utilize TeA, the conditions of TeA production by Alternaria alternata(No.: 5390) were studied. The effects of different media, temperature, time and illumination on TeA content were studied by single factor experiment. On this basis, response surface was used to optimize the production conditions of A. alternata. The results show that the optimal conditions were Richards medium, dark culture, temperature 30 ℃ for 29 days. Under these conditions, the TeA content was 14.99 μg/g, which was close to the model prediction. Screening the culture conditions of A. alternata producing TeA and exploring the method of obtaining A. alternata toxin TeA by fungus culture will be beneficial to the further development and utilization of TeA.