The mutation of Opaque2 (O2) significantly affect protein body formation, starch content, and amino acid composition in the endosperm of maize kernels, and the increasing of lysine content significantly improves nutrition quality. Through the backcrossing scheme, we constructed near isogenic lines of 17 waxy maize backgrounds with the o2 mutation gene. By analyzing the trait of the kernel, including appearance, pericarp thickness, 100-kernel weight and crude starch content, we found that the kernels of 16 near isogenic lines were shrunken significantly and their endosperm changed to opaque obviously compared to the controls. Pericarp thickness of 6 lines increased, and the 100-kernal weight of 11 lines decreased significantly, crude starch content of 12 lines decreased significantly. It indicated that the introduction of o2 mutant gene mainly have a negative effect on kernel appearance, yield and crude starch content with waxy maize, but the degree of influence is related to maize background.