As an important indicator of forage production, aboveground biomass will directly illustrate the growth of forage grass. Therefore, Real-time monitoring biomass of forage grass play a crucial role in performing suitable grazing and management in artificial and natural grassland. However, traditional sampling and measuring are time-consuming and labor-intensive. Recently, development of hyperspectral remote sensing provides the feasibility in timely and nondestructive deriving biomass of forage grass. In the present study, the main objectives were to explore the robustness of published and optimized spectral indices in estimating biomass of forage grass in natural and artificial pasture. The natural pasture with four grazing density (control, light grazing, moderate grazing and high grazing) was designed in desert steppe, and different forage cultivars with different N rate were conducted in artificial forage fields in Inner Mongolia. The canopy reflectance and biomass in each plot were measured during critical stages. The result showed that, due to the influence in canopy structure and biomass, the canopy reflectance have a great difference in different type of forage grass. The best performing spectral index varied in different species of forage grass with different treatments (R2=0.00~0.69). The predictive ability of spectral indices decreased under low biomass of desert steppe, while red band based spectral indices lost sensitivity under moderate-high biomass of forage maize. When band combinations of simple ratio and normalized difference spectral indices were optimized in combined datasets of natural and artificial grassland, optimized spectral indices significant increased predictive ability and the model between biomass and optimized spectral indices had the highest R2 (R2=0.72) compared to published spectral indices. Sensitive analysis further confirmed that the optimized index had the lowest noise equivalent and were the best performing index in estimating biomass. In conclusion, optimizing wavebands combination was a promising algorithm for improving prediction abilities of biomass for forage grass.