At present there are many methods to discuss the crystallinity of lignocellulose but scholars experts have different opinions about them. This study, with bamboo under the different pretreatment conditions, and with help of acid, alkali and glycerol uses X-ray diffraction and CP/MAS13NMR in order to make clear of the crystallinity of bamboo change characteristics, and measured the area length, spacing, crystal plane diffraction peak position, the crystallinity index and parameters and complete the comparison of crystalline cellulose and Gaussian function curve C - 4 area signal area. Results showed that crystalline overall increased significantly after pretreatment, 002 peak become sharp and offset to the direction of large angle for alkali pretreatment, 002 crystal width increases, crystalline spacing is smaller. CP/MAS13NMR is consisting with XRD, but calculated value is smaller. It proved that effective processing can effectively disrupted the recalcitrance of bamboo, and produce highly active cellulose in lignocelluloses’ biomass ethanol production, that provide guidance to realize the lignocellulose bioconversion and effective utilization.