The invention relates to a tissue culture and rapid propagation method of acer palmatum. The method comprises the following steps of: after a series of sterilization processes, carrying out isolated culture on acer palmatum stems by adopting NN69 as a basic culture medium; and establishing a rapid propagation system of the acer palmatum by utilizing hormone regulation and control to successfully culture an integrated rooting regeneration plant with a transplant survival rate of about 90 percent, wherein a starting culture medium for inoculating a primary generation comprises the NN69, 0.1mg/L of NAA (Naphthylacetic Acid), 0.3mg/L of TDZ (Thidiazuron) and 30g/L of sucrose with a pH value of 5.8; an enrichment culture medium comprises the NN69, 30g/L of sucrose, 0.1mg/L of NAA and 0.8mg/L of KT-30; and a rooting culture medium comprises 1/2 NN69, 0.2mg/L of NAA, 20g/L of sucrose and 0.2g/L of active carbon. The method overcomes the defects of long conventional seedling culturing period of the acer palmatum, low cuttage propagation coefficient, and the like and realizes the industrial production of acer palmatum nursery stocks.