The efficient conversion of lignocellulose relies on the implementation of an effective pretreatment strategy. Cationic surfactants combined with deep eutectic solvent ([Betaine][LA] was employed as a novel pretreatment strategy for treating corn stalks. Valuable insights were provided on the impact of the surfactant hydrophobic segment length on pretreatment efficacy. The specific pretreatment conditions were optimized by single factor and orthogonal experiment. Octadecyl trimethyl ammonium bromide (OTAB, 1.5 wt%) with the longest hydrophobic part combined with [Betaine][LA] (Betaine-to-LA molar ratio 1:4) achieved the best pretreatment effect (delignification 92.8 %, xylan elimination 91.1 %) when severity factor reached 4.26, meanwhile, 1.7 g/L xylo-oligosaccharides and 4.4 g/L furfural were detected in pretreatment liquid due to the hydrolysis of hemicellulose in corn stalks with acidic deep eutectic solvent [Betaine][LA]. The relative saccharification activity reached 2.7 times of raw material, while biomass crystallinity, lignin surface area and lignin hydrophobicity significantly decreased, leading to enhanced cellulose accessibility. Additionally, molecular perspective provided by molecular dynamics showed the elimination of lignin and hemicellulose was facilitated by strong interaction of hydrogen-bond and van der Waals force between lignin and hemicellulose with [Betaine][LA] + OTAB. Overall, the effectiveness and potential of cationic surfactant combined deep eutectic solvent pretreatment strategy for lignocellulose pretreatment was revealed.