Ni-based electrocatalysts with strong redox abilities are active for the electrochemical oxidation of 5-hydroxymethylfurfural (HMF).Interface engineering is an efficient way to modulate the electronic structure,tune the intermediate adsorption,and expose more active sites.Herein,we increased the concentration of interfacial sites with rich defects in a 3D hierarchical nanostructured NiO-Co3O4 electrocatalyst and investigated its catalytic performance for HMF electro-oxidation.The interface effect created abundant cation vacancies,modulated the electronic properties of Co and Ni atoms,and raised the oxidation state of Ni species.The NiO-Co3O4 catalysts show superb HMF oxidation activities with a low onset potential of 1.28 VRHE.Meanwhile,in-situ surface-selective vibrational spectroscopy of sum-frequency generation was performed to study the reaction pathway during the oxidation process on the electrocatalysts.The current study offers an efficient way to create cation vacancies and proves the decisive role of cation vacancies in catalyzing the HMF electro-oxidation.