Culture of soybean mesophyll protoplasts in alginate beads

• Published in: Plant cell reports Vol. 5; no. 5; pp. 334 - 337
• Main Authors: Tricoli, D.M, Hein, M.B, Carnes, M.G
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01-10-1986
• Subjects: Agronomy. Soil science and plant productions; Biological and medical sciences; Biotechnology; cell culture; Cell cultures. Hybridization. Fusion; Economic plant physiology; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; Glycine max; growing media; In vitro culture; methodology; Methods. Procedures. Technologies; Molecular and cellular biology; Plant cells and fungal cells; Protoplast preparation, cell fusion and regeneration; protoplasts; Cell culture; Vegetals; Leguminosae; Dicotyledones; Protoplast; Angiospermae; Cell division; Spermatophyta; Alginates; Fodder crop; Glycine max; Oil plant(vegetal)
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/BF00268595

Mesophyll protoplasts were isolated from leaves of 10 day old aseptically grown soybean seedlings, or from surface disinfested leaves of 3 week old plants grown in environmental chambers. The protoplasts were encapsulated in 2mm diameter Ca alginate beads. Immobilized protoplasts were induced to divide by culturing in shaker flasks containing an actively growing soybean cell suspension. The feeder cell suspension supported the division of protoplasts independent of the protoplast density in the Ca alginate beads. At day 18 after encapsulation, the alginate matrix was dissolved, releasing viable callus colonies. The feeder cell suspension obviated plating of protoplasts at high density which is usually required for subsequent cell division and colony development. Since the protoplasts were embedded at low density, the cell colonies were derived from single cells.