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PJB-2002-42
OPTIMIZATION OF CONDITIONS FOR ELECTROFUSION IN SUGARCANE PROTOPLASTS
Abstract
Somatic hybridization studies were performed using leaf mesophyll as well as homogeneous cell suspension-derived protoplasts in sugarcane (Saccharum spp. hybrid cvs. CoL-54 and CP-43/33). The conditions standardized for electrofusion were 40 V of primary voltage for 30 seconds of AC for pearl chain formation and 2.0 KV cm-1 for 20 mS in DC, 4 times with repeat intervals of 5S for fusion. About 60% of the protoplasts underwent fusion event (pearl chain formation) and single pair fusion was observed in about 21% of the protoplasts. Microcalluses from such protoplasts were successfully achieved on KM8P-K8P medium after 20 days of culture.
