The effects of leaf spraying and root irrigation with amino acids water soluble fertilizer and EM microbial agents on the growth, photosynthetic characteristics, yield and quality of lettuce were studied. 4 treatments were designed in the experiment: CK, no fertilizer was applied; T1, conventional fertilizer; T2, EM microbial agent; and T3, amino acids water soluble fertilizer. The results showed that compared with CK, spraying + irrigating EM microbial agents and spraying + irrigating amino acids water-soluble fertilizer could significantly increase the plant height and stem diameter of lettuce, and effectively promote the growth of lettuce. Compared with T1, the photosynthesis of lettuce was also significantly improved by spraying and irrigating amino acids water-soluble fertilizer. Lettuce yield was in order of T3 > T2 > T1 > CK, T1, T2 and T3 increased lettuce yield by 20.17%, 26.05% and 31.65%, respectively; T2 and T3 increased lettuce yield by 4.90% and 9.99%, respectively compared with T1, and T3 increased lettuce yield by 4.44% compared with T2, which indicated that amino acids water soluble fertilizer and EM microbial agents could increase lettuce yield, and the effect of amino acids water soluble fertilizer is better.