Branches from three transgenic poplar lines and their wild type line 107 were used to study the effect of lignocellulosic composition and structure on the production of glucose and ethanol. Experimental results showed that the transgenic line 18-1 had the high cellulose content and amorphous fibril structure. After poplar meals were pretreated with 10% NaOH and a mixture of hydrogen peroxide and acetic acid, their lateral order index decreased significantly. The highest glucose yield in enzymatic hydrolysis and ethanol yield from the substrate of 18-1 was much higher than that from feedstock of 107 by 192.7% and 108.7%, respectively. Scanning electron microscopy images confirmed that lignocellulose from the 18-1 could be destroyed by chemicals more easily than those from other lines. These results demonstrated that changing lignocellulose structure could be more effective on improving the digestibility and enzymatic hydrolysis of poplar biomass than increasing the cellulose content in biomass.