Abstract
Proroplasts were enzymatically isolated from protonema of Funaria hygrometrica L. SIBTH., cultured from the spores on Knop's solid medium. The protoplasts were incubated in hanging drops in liquid Knop's medium with 0.3 M mannitol and 0.5% glucose. Isolation of protoplasts was obtained successfully, through the action of Macerozyme®, cellulase and Rohament P® mixed with mannitol. Cultured protoplasts began to regenerate walls within 24 h, followed by the enlargement and divisions within 48 h. These protoplasts started producing active and long strands of protonema within 5 days, while well-differentiated leafy gametophytes were developed just in 20–23 days.
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