Rice straw can be used for biochemical conversion for energy production. The use of near infrared spectroscopy was explored to estimate the contents of cellulose, hemicellulose, and lignin in this study. A total of 172 samples from 17 provinces of China were collected and scanned using NIRSystem 6500 spectrophotometer. Different spectra pretreatments and regression methods were adopted to optimize the models. When evaluated on independent validations, the standard errors of validation and determination coefficient of validation were 1.49%, 1.59%, 0.65% and 0.82, 0.71, 0.78, respectively. It was concluded that near infrared spectroscopy can be used to roughly analyze the contents of cellulose and lignin, and can only distinguish high contents and low contents of hemicellulose in rice straw.