To decrease fertilization amount of chemical fertilizer and improve the quality of vegetable crops, spinach was taken as the test material, and the impact of different fertilizer on spinach growth and yield was studied via the manners of biological organic fertilizer and organic fertilizer+chemical fertilizer. Experimental results showed that in the formula of organic fertilizer+chemical fertilizer, chlorophyll and nitrogen contents in spinach leaves obviously increased; in the formula of only adding organic fertilizer, spinach leaf temperature, leaf width, root length, plant height and fresh weight were all better than those in the formula of organic fertilizer+chemical fertilizer, and better formulas were A5, E5, F3 and I5, in which spinach plant height in E5 was 5.63 times higher than G5, root length in E5 was 2.67 times higher than G5, and fresh weight in G5 was 32.6 times higher than G5. By comprehensive analysis, the most suitable formula for spinach production was E5, and the research could provide theoretic basis for fertilization amount of organic fertilizer required by spinach growth and development.