Abstract: The effects of exogenous auxin（NAA and IBA） concentration and different substrate types on the rooting index and rooting quality of semi-lignified shoots and cuttings of Acer mono（A. mono） was studied to select the best hormone concentration and substrate to lay the foundation for breeding and promote cultivation of A. mono. The semi-lignified A. mono cuttings of the same year were used as materials, and the concentrations of five kinds of combined solutions of NAA and IBA（NAA+IBA） were set at 0, 500, 1 000, 1 500 and 2 000 mg/L. A single factor randomized block test was carried out on four substrates of vermiculite, peat soil, perlite, perlite+vermiculite. Results showed that: different NAA+IBA concentrations and substrate types had extremely significant effects on the measurement indexes（P＜0.01）, and all had the highest rooting rate on the 40 th day. Among different hormone concentrations, the rooting rate（62.50%）, callus induction rate（66.67%） and total length of new roots of 1 500 mg/L treatment were the best. Comparing the total number of roots, it was found that there was no significant difference between 1 500 and 2 000 mg/L, but both were significantly better than 0, 500, 1 000 mg/L treatments. Among the four matrices, perlite performed the best in the rooting rate（66.67%） and callus induction rate（70.84%）; perlite+vermiculite（1∶1） had the best performance in the total length and total number of new roots. The A. mono cuttings was significantly affected by the concentration of NAA+IBA and different substrate types: considering the rooting effect and rooting quality, 1 500 mg/L was the best concentration of NAA+IBA hormone; perlite was significantly better than other substrates in terms of rooting index, which was the best choice; in the later stage of rooted cuttings, if the quality of rooting was pursued, perlite+vermiculite should be used as the cultivation substrate. The cutting method established in this paper can be used for the expansion and reproduction of excellent germplasm resources.