Design and preparation of frequency doubling antireflection coating with different thicknesses of interlayer were investigated for LiB3O5 (LBO) substrate. The design was based on the vector method. The thickness of the inserted SiO2 interlayer could be changed in a wide range for the four-layer design with two zeros at 1064 and 532 nm. The coatings without any interlayer and with 0.1 quarter-wave (QW), 0.3 QW, 0.5 QW SiO2 interlayer were deposited respectively on LBO by using electron beam evaporation technique. All the prepared coatings with SiO2 interlayer indicated satisfying optical behavior. This expanded our option for the thickness of an interlayer when coating on LBO substrate. The prepared films with SiO2 interlayer showed better adhesion than that without any interlayer. The thickness of the interlayer affected the adhesion, the adhesion for the coating with 0.5 QW SiO2 interlayer was not as good as the other two.