Plant Tissue Culture 1 (1) : 1-8, June (1991)

ANALYSIS OF THE FUSION PROCESS OF PLANT PROTOPLASTS BY FLUORENCE MICROSCOPY

Takeshi Taniguchi and Eizo Maeda

Faculty of Agriculture, Nagoya University, Chikusa, Nagoya-464,Japan

Key words:

Abastract

When protoplasts from rice callus and lettuce cotyledons were stained with rhodamine 123 (R123), many yellow spots (mitochondria) were observed. After incubation in 0.5M mannitol solution for one day, the fluorescence intensity of mitochondria diminished or disappeared. By the viability test with Evans blue or fluorescein diacetate, almost all of the protoplasts of rice and lettuce were viable immediately after their isolation but less than 50% after one day culture in 0.5M mannitol. When FITC (fluorescein isothiocyanate) ? stained lettuce protoplasts were fused with the non-stained protoplasts, long-shaped fusion bodies were formed, and finally some became round-shaped ones. During the process, FITC moved into non-stained areas. FITC-, R123- or phenosafranine ?stained rice callus protoplasts were fused with rice callus or shoot protoplasts. The fusion process proceeded very rapidly and sometimes produce very big fusion bodies. The inside of the fusion bodies was not homogeneous and some round-shaped particles were observed. FITC-stained rice callus protoplasts were fused with lettuce protoplasts. Chloroplasts of lettuce moved into lettuce protoplast areas. The speed of the fusion was higher than that between lettuce protoplasts.

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