The invention discloses a Decaisnea insignis isolated culture method. The method comprises that an immature fruit seed embryo as an explant is orderly subjected to callus induction, adventitious bud differentiation induction, adventitious bud enrichment culture and rooting induced culture so that a Decaisnea insignis regeneration plant is obtained. Before callus induction, the method also comprises explant disinfection: taking tender Decaisnea insignis fruits, immersing the tender Decaisnea insignis fruits in a cleaning solution for 20-30min, wherein each 100ml of the cleaning solution contains 6-10 droplets of a cleaning agent, then scrubbing the surfaces of the Decaisnea insignis fruits, washing the Decaisnea insignis fruits through water for 1-2h, carrying out disinfection treatment on the tender Decaisnea insignis fruits through alcohol with a concentration of 75% in an aseptic environment for 30-60s, carrying out disinfection on the Decaisnea insignis fruits through a HgCl2 solution with a concentration of 0.1% for 15min, wherein each 100ml of the HgCl2 solution contains 3-4 droplets of Tween-40, cleaning the Decaisnea insignis fruits through sterile water 4-6 times, for 1-3min every time, taking out tender seeds, extruding tender seed embryos and rinsing the tender seed embryos through sterile water to obtain explants. The method realizes high frequency induction of a lot of Decaisnea insignis clustered adventitious buds and has a high transplanting survival rate. The method lays a good foundation of Decaisnea insignis propagation coefficient improvement and specie germplasm preservation, promotion and use.