Hyperspectral imaging (HSI) technology has increasingly been applied as an analytical tool in fields of agricultural, food, and Traditional Chinese Medicine over the past few years. The HSI spectrum of a sample is typically achieved by a spectroradiometer at hundreds of wavelengths. In recent years, considerable effort has been made towards identifying wavelengths (variables) that contribute useful information. Wavelengths selection is a critical step in data analysis for Raman, NIRS, or HSI spectroscopy. In this study, the performances of 10 different wavelength selection methods for the discrimination of Ophiopogon japonicus of different origin were compared. The wavelength selection algorithms tested include successive projections algorithm (SPA), loading weights (LW), regression coefficients (RC), uninformative variable elimination (UVE), UVE-SPA, competitive adaptive reweighted sampling (CARS), interval partial least squares regression (iPLS), backward iPLS (BiPLS), forward iPLS (FiPLS), and genetic algorithms (GA-PLS). One linear technique (partial least squares-discriminant analysis) was established for the evaluation of identification. And a nonlinear calibration model, support vector machine (SVM), was also provided for comparison. The results indicate that wavelengths selection methods are tools to identify more concise and effective spectral data and play important roles in the multivariate analysis, which can be used for subsequent modeling analysis.