Green (GV) and non-photosynthetic vegetation (NPV) cover are both important biophysical parameters for grassland research. The current methodology for cover estimation, including subjective visual estimation and digital image analysis, requires human intervention, lacks automation, batch processing capabilities and extraction accuracy. Therefore, this study proposed to develop a method to quantify both GV and standing dead matter (SDM) fraction cover from field-taken digital RGB images with semi-automated batch processing capabilities (i.e., written as a python script) for mixed grasslands with more complex background information including litter, moss, lichen, rocks and soil. The results show that the GV cover extracted by the method developed in this study is superior to that by subjective visual estimation based on the linear relation with normalized vegetation index (NDVI) calculated from field measured hyper-spectra (R2 = 0.846, p < 0.001 for GV cover estimated from RGB images; R2 = 0.711, p < 0.001 for subjective visual estimated GV cover). The results also show that the developed method has great potential to estimate SDM cover with limited effects of light colored understory components including litter, soil crust and bare soil. In addition, the results of this study indicate that subjective visual estimation tends to estimate higher cover for both GV and SDM compared to that estimated from RGB images.