The invention discloses a preparation method of cellulosic ethanol, which comprises the following steps of pulverizing lignocellulosic biomass, carrying out hydrothermal reaction, cooling to obtain a mixed solution, regulating the pH value, adding cellulase and xylanase, carrying out enzymolysis, inoculating yeast, and fermenting to prepare cellulosic ethanol. According to the method, hydrothermal short-time depolymerization pretreatment is utilized to obtain the low-toxicity pretreatment liquid, and the cellulosic ethanol is prepared by a one-pot method, so that the operation steps are simplified, xylan in the pretreatment liquid is retained, pentose and hexose co-fermentation is realized, the yield of the cellulosic ethanol is remarkably increased, and the method is low in cost, low in energy consumption, free of wastewater discharge and effective sugar components in lignocellulose can be comprehensively utilized.