Abstract: Effects of sulphureous fertilization on photosynthesis, physiology and yield characteristics of two winter wheat (Triticum aestivum L.) cultivars (Yumai 49 with a multiple spike and Yumai 66 with a heavy spike) were systematically studied in two successive years in the field. The results show that the sulphur fertilization plays an important role in improving canopy apparent photosynthesis, flag leaf’s net photosynthesis, leaf’s chlorophyll content and flag leaf’s nitric acid reductase activity. However, there are differences among the different sulphur fertilizer treatments, and as well as between the two cultivars. In addition, the study also indicates that there are differences in increasing grain yield and improving yield characteristics of the two cultivars among the different sulphureous fertilizer application. On the whole, compared with photosynthesis, physiology and yield characteristics and input cost during wheat growth stage, the rate of S fertilizer recommended presently is S 120 kg/ha as base fertilizer application for the cultivars under the high-yield wheat field conditions in Henan province of China.