A study on isolation and culture of pollen protoplasts

Abstract

Protoplasts were isolated from mature pollen grains in Iris tectorium; Zephyranthes grandiflora and Hemerocallis fulva by an enzymatic technique. The protoplasts were purified by discontinuous gradient centrifugation and confirmed to viable by fluorochromatic reaction. When the protoplasts of H. fulva were cultured, the regeneration of cell wall, formation of various tube-like structures and sometimes division of the generative nucleus into 2–4 nuclei took place. Protoplasts from tetrads to mature pollen except early microscope stage were obtained in H. fulva as well.