Abstract: In order to study influence of drought stress on physiological, biochemical and yield of soybean, 22 different eco-types soybean cultivars were used as materials to test drought-resistant characteristics and physiological response in the conditions of pot experiments and field tests with normal water supply and water stress. The result showed that, among the 22 cultivars, ‘Jinda74’ had the strongest drought resistance and ‘Bianjing’ had the weakest drought resistance. Under water stress, relative water content, activity of POD and SOD, chlorophyll a content, total chlorophyll content and net photosynthetic rate declined. The cultivars with strong drought resistance had a smaller decline ranges than the weak. The indexes were positively correlated with drought resistance, the correlation coefficient were respectively r=0.8455, r=0.9764, r=0.8776, r=0.9118, r=0.8803, r=0.8312. MDA content, relative conductivity and soluble sugar content increased. The cultivars with strong drought resistance had smaller increasing rages while the cultivars with weak drought resistance had greater increasing rage. These indexes were negatively correlated with drought resistance, the correlation coefficient were respectively r=-0.7847, r=-0.8726, r=-0.7025. Under drought stress, Pro has a very significant positive correlation (r=0.8117) with drought resistance, it had a larger increase in stronger drought resistance varieties. The results indicated that water deficit caused RWC in soybean leaves reduced, lipid peroxidation damage increased, MDA accumulation, RC increased, Pro and SS increased. MDA content was responsible for declining activity of protecting enzymes. At the same time, water deficit resulted in declining content of chlorophyll, photosynthetic rate and biomass. The stronger drought-resistant cultivars not only have higher yield under normal water supply but also get a stable yield under drought stress.