Abstract: Root-controlling containers are used to cultivate seedlings in this research, and 13 mediums made from plant straw, orchard soil, peat soil, perlite and cow dung are added in the containers. Effects of different mediums on growth, rooting and leaf photosynthesis of 2-year-old apple seedlings are studied compared with seedlings with orchard soil. Results showed that the seedling height, stem diameter, length and diameter of new shoots, dry matter weight of the aerial part and root development of seedlings grown in the 13 mediums were all better than those of the control. Furthermore, net photosynthetic rate, transpiration rate, stomatal conductance and intercellular CO2 concentration were also higher than those of the control. On the other hand, the rootshoot ratio, the average diameter of root and the water use efficiency were lower than those of the control. It was also found that the aerial part growth, rooting and root growth, as well as leaf photosynthesis of seedlings grown in medium No.6 (orchard soil: peat soil: cow dung: perlite=2:3:1:1) and No.7 (orchard soil: corn straw: cow dung: perlite=2:3:1:1) were obviously better than those of seedlings grown in other mediums. All the above results reveal that medium No.6 and No.7 could meet the nutrient requirement of young apple trees during growth and development. In particular, medium No.7 is more economical and practical, which can be used as an excellent culture substrate for apple seedlings.