The Effect of High pH and Calcium on Tobacco Leaf Protoplast Fusion

Abstract

Abstract High frequencies of fusion were induced between isolated tobacco leaf protoplasts when they were incubated at 37 °C in the presence of 0.05 M CaCl 2 in 0.4 ᴍ mannitol at pH 10.5. Subsequent to the fusion treatment the protoplasts were washed and cultured in a suitable medium and within two weeks, actively dividing cell colonies were observed. The fusion treatment had no irreversible, deleterious effects on the protoplasts