Cornstalk is a typical cellulose material, which can be used by photo-fermentative H-2 production after pretreatment. However, the pretreatment methods have different influence on photo fermentation. In this study, 25.0 g cornstalk was pretreated by HCl/NaOH/cellusase. The hydrolysis rates increased from 45.51% by ddH(2)O-treatment to 60.79% by diluted HCl-treatment and 51.6% by NaOH-treatment. The corresponding reducing sugar yields were 0.13 g/g, 0.42 g/g and 0.01 gig, respectively. Enzymatic treatment enhanced the corresponding cornstalk hydrolysis rates to 50.81%, 67.60% and 64.10% with reducing sugar yields of 0.22 g/g, 0.62 g/g and 0.26 g/g. The sorts and concentrations of carbon source for H-2 production vary among different hydrolysates. Photo-fermentative H-2 production of strain R. capsulatus JL1 and mutant JL1601 (cheR2(-)) with hydrolysates were investigated. The maximum H-2 yield of 123.8 +/- 14.2 mL/g by strain JL1 was obtained from alkali-enzyme pretreated cornstalk, while the H2 yield of 224.9 +/- 5.2 mL/g by mutant JL1601 (cheR2(-)) was obtained with acid-enzyme hydrolysate as the substrates. Meanwhile, the alkali pretreated cornstalk was the worst for photo-fermentation of both strain JL1 and mutant JL1601 (cheR2(-)). Nevertheless, the highest substrate conversion efficiencies for both strains were obtained from ddH(2)O-pretreated hydrolysate. Two-step pretreated hydrolysates were more beneficial to H-2 production for mutant JL1601 (cheR2(-)) but not for strain JL1. (C) 2019 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.