The aim of this work was to study the ability of NIR spectroscopy to determine holocellulose and alpha-cellulose contents in triploid clones of Populus tomentosa. For this purpose 90 trees with five families in different planting density grown in Shandong were analyzed. Calibration models were developed from wet chemical and NIR data using partial least squares (PLS) regression. High coefficients of determination (R2) and low root mean square errors of cross-validation (RMSECV) were obtained for holocellulose (R2 = 0.7805, RMSECV =0.652) and alpha-cellulose (R2 = 0.8702, RMSECV = 0.548) from wood meal. Prediction produced high correlation coefficients between laboratory and predicted values for holocellulose and alpha-cellulose contents, which R2 values are 0.7467 and 0.8505, and RMSEP are 0.993 and 0.553, respectively. This study showed that NIR analysis can be reliably used to predict holocellulose and alpha-cellulose contents in triploid clones of Populus tomentosa.