Although biological pretreatment has the advantages of being environmentally friendly and having low-energy consumption, it usually requires a relatively long incubation time. In this study, a novel combined pretreatment with white-rot fungus and alkali at near room-temperature for saccharification of corn stalks was investigated to speed up the biological process. Biological pretreatment with Irpex lacteus or Echinodontium taxodii can improve enzymatic hydrolysis of corn stalk greatly, but the process requires a long time (60 days) to achieve a satisfactory sugar yield. The combination processes with the fungi were compared with the sole pretreatments. The results showed that the time of the biological process could be shortened to 15 days when the bio-treatment with I. lacteus was combined with alkali pretreatment. The efficiency of alkali pretreatment can be also enhanced significantly by biological treatment. 271.1mg/g of final glucose yield was obtained for the combination pretreatment, which was an improvement of 50.4% and 28.3% in comparison with the sole alkali pretreatment at the same and optimum reaction time, respectively. In conclusion, the combination of biological pretreatment with alkali processes not only speeded up the biological process, but also improved the sugar yield in comparison to the sole pretreatment and is favorable for the development of biological pretreatment.