Abstract:An osmotic shock method of isolating generative cells from Allemanda neriifolia was described. Fresh pollen grains were first placed ill a Brewbaker and Kwack‘s medium (BK medium) containing 50% sucrose, incubated at 28℃ for 2 hours. During this incubation period pollen grains germinated and produced pollen tubes measuring about 200 μm long. After this initial incubation period, a fixed amount of BK medium without sucrose was added thus diluting the original medium to a sucrose concentration of 30% – an optimum concentration for generative cell growth. The addition, of the BK medium without sucrose brought about an osmotic shock effect on the pollen tubes and caused most of the tubes to burst at the tip region thus releasing the contents together with the generative cell from the tube into the 30% sucrose + BK medium. After isolation and filtering into a fresh lot of 30% sucrose + BK medium, generative cells changed from spindle into spherical-shaped cells. In the 30% sucrose + BK medium, the generative cells divided and within a short period of 3 to 5 hours a laege number of cells at various stages of mitosis was obtained.