In this experiment, five substrate formulas with sand, sawdust, peat and bio–charcoal were conducted to study the effect of bio–charcoal on the growth of Poncirus Trifoliata (L.) Raf. with container nursery. The results showed that the bio–charcoal was alkaline, its pH was as high as 8.95 and could be used to improve pH of mix substrates. It was rich in P and K and lack of N. The bio–charcoal could effectively improve the water holding capacity, pH and mineral elements content, reduce bulk density and increase the capillary porosity in mix substrates of container nursery, but the impact on conductivity and cation exchange capacity were not obvious. The substrate formula with bio–charcoal had obvious effect on Poncirus trifoliata growth, its plant height was higher than CK1, CK2, but the difference between them was no significance. The root fresh weight and dry weight were higher than CK2 as well.