| Popis: |
An optimized protocol was established to isolate large numbers of mature living generative cells from Nicotiana tabacum, Vicia faba L., and Iris tectorium Maxim. Fresh pollen grains on the day of anthesis were incubated in a solution containing 9% mannitol. The optimal pH and temperature for the reaction were 5.8 and 24°C, respectively. For a short time, large amount of generative cells were subsequently released into the solution and were immediately collected for preparation. The isolated pollen protoplasts were demonstrated to be living by microscopy in a fluorochromatic reaction with fluorescein diacetate (FDA). The isolation rate was above 95%. |
