Abstract: 【Objective】 In vitro embryo culture can effectively shorten the time of crop breeding, especially in the generation-adding technique of cotton breeding, which has important application potential. This project aims to study the effects of different factors on seedling formation of cotton excised embryos, and explore the combination of key factors in the hope of providing theoretical basis for the large-scale cultivation of cotton excised embryo. 【Method】 The effects of culture mediums, embryo ages, temperatures and illumination times on cotton embryo seedling formation were studied to analyze embryo seedling rate, seedling time and the development characteristics of root, in order to make clear the suitable and simplified cultivation conditions. 【Result】 Among different culture media, MS(Murashige and Skoog)medium had the highest seedling rate, followed by tap water and distilled water. With the increase of embryo age, the seedling rate increased gradually; when the embryo age was 35 d (day), there was no significant difference among the three culture medium. Among different embryo ages, the seedling rate of 35 d was the highest, and the seedling time was the shortest. When the embryo age was 35 d, 30℃ was the most suitable temperature for shortening seedling time and promoting root development. Illumination time 24 h (hour) was beneficial to shorten seedling time, but was not conducive to root development. 【Conclusion】 Culture medium, embryo age, temperature and illumination time are the key factors that affect the seedling formation of cotton excised embryo. Advanced embryo age, suitable temperature and illumination time are beneficial to the seedling formation of excised embryo, shortening seedling time and promoting root development. Under the conditions of embryo age 35 d, temperature 30℃ and illumination time 24 h, the effect of hydroponics of excised embryo is good, which can be used as the suitable condition of simplified seedling culture of early-maturing upland cotton excised embryo.