The invention discloses an ex-vitro rooting method for 
tissue culture seedlings of 
pinus massoniana. The ex-vitro rooting method comprises the following steps: taking terminal buds of aseptic seedlings with 0.5cm long hypocotyls as explants for inducing cluster buds in a culture medium I; carrying out elongation culture on the induced cluster buds in a culture medium II, individually 
cutting off the terminal buds, and inoculating the 
cut terminal buds into a culture medium III for propagation to obtain subculture buds I; repeatedly elongating the subculture buds I, individually 
cutting off the subculture buds I, dividing the 
cut subculture buds I into terminal buds and 
bud sections, continuously propagating in a subculture propagation culture medium III to obtain subculture buds II, and repeatedly elongating and propagating the subculture buds II to obtain new propagation subculture buds; and 
transplanting the individually 
cut 2.5-3.5cm high subculture buds into a 
seedling culture cup for ex-vitro rooting, and 
transplanting the ex-vitro subculture buds into a 
seedling nursery for culture to obtain the ex-vitro 
tissue culture seedlings. According to the method disclosed by the invention, the production program of test-tube seedlings is simplified, the production cost is reduced, the production efficiency is improved, the rooting 
treatment time is 25-30 days, the ex-vitro rooting rate reaches 92%-98%, the 
root system quality is higher, the 
transplanting survival rate reaches 90%-95%, and the method has better economic, social and ecologic benefits.