To better understand the nature of reactive adsorption of thiophene on Ni/ZnO adsorbent, the effects of ZnO textural structure on the desulfurization activity were investigated. Flower-like ZnO materials were synthesized by hydrothermal homogeneous precipitation method and the corresponding Ni/ZnO adsorbents were prepared by incipient impregnation method. The analysis results showed that both the crystalline sizes and the BET surface areas of flower-like ZnO were better than that of commercial ZnO. The activity evaluations indicated that the Ni/ZnO adsorbents prepared with ZnO possessing a favorable textural structure exhibited good activity of removing thiophene. The evolutions of the main crystalline phases of Ni/ZnO adsorbents before and after reaction confirmed that ZnO played a crucial role in taking up S element and converted it into ZnS in the reactive adsorption process. It was concluded that ZnO with larger surface area and smaller crystal particles resulted in better desulfurization activity, which may be the main reason for the better activities of the Ni/ZnO adsorbents prepared with flower-like ZnO matterials.