This study tested the effect of hydrothermal treatment (HTT) at different peak temperatures (150 degrees C and 210 degrees C, i.e. HTT150 and HTT210, respectively) and holding time (0-30 min) on the solubilization of rice straw at total solids (TS) of 20% and then subsequent H-2 production from resultant substrates. No obvious degradation was detected in lignin content under all tested HTT conditions which did open up the surface structure and have efficient solubilization effect on rice straw. Soluble carbohydrates produced from straw particles during HTT210 was found to have strongly (r = 0.9987) positive correlation with the subsequent H-2 yield. The maximum soluble carbohydrates, 80 mg per gram of volatile solids (VS) was achieved under HTT210 and 0 min of holding condition, correspondingly yielding the highest hydrogen production (28 ml/g-VS), about 93-fold higher than the control. Results suggest that holding time is crucial for HTT pretreatment when taking subsequent H-2 production into consideration. (C) 2014 Elsevier Ltd. All rights reserved.