| Abstract | Protoplasts were isolated from shoot tips of 3-day-old seedlings of Pisum sativum L. var. Century by enzymatic degradation of the cell walls. The protoplasts originated from cells of the meristematic domes and parenchymatous tissues of 2- 3-mm tip sections using two consecutive steps of enzyme treatment in the presence of hexitols and calcium salts. Upon culturing, wall regeneration had occurred after 24 h, and cell division was observed within 3–5 days. Supplementing the culture medium with 2−5 mMl-glutamine, 5−10 μM kinetin, 1−5 μM dichlorophenoxyacetic acid (2,4-D) and 0.1−0.5 μM naphthaleneacetic acid (NAA) resulted in sustained cell division and callus formation in five weeks. Glucose at 0.3 M was superior to sorbitol and mannitol as osmotic stabilizers. |
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