In this study, three cuttings, 1-year-old with terminal bud, 1-year-old without terminal bud and 2-year- old hard branches of Acer rubrum ‘October Glory’, each were applied four types of cutting substrates, named M1 (equal volume of sand and perlite), M2 (equal volume of sand and vermiculite), M3 (equal volume of perlite and vermiculite) and M4 (equal volume of sand and perlite and vermiculite) separately, as well as applied IBA and NAA at different mass concentration ratios. The effects of cutting type, substrate type and exogenous hormones on rooting, the histological observation of adventitious root formation during cutting process were investigated. Results showed that M4 matrix was the most suitable cutting substrate showing the best rooting rate (69.44%), average rooting number (14.31) and average root length (5.40 cm). The 1-year-old cuttings showed higher rooting rate and average number of roots but lower average root length than the 2-years-old cuttings. And, the difference of rooting rate between 1-year-old with terminal bud and 1-year-old without terminal bud cuttings was not significant (P<0.05). The combination of 500 mg/L IBA+500 mg/L NAA was the best application in rooting of cuttings,with the rooting rate 75.56%, the average rooting number 15.68, and the average root length 6.48 cm. The adventitious roots of Acer rubrum are derived from the cross areas of the xylem rays with the vascular cambium and phloem parenchyma, which belongs to the induced root primordia of multilocus type.