Microcallus formation from Hevea brasiliensis protoplasts isolated from embryogenic callus

• Published in: Plant cell reports Vol. 13; no. 5; pp. 272 - 276
• Main Authors: CAZAUX, E, D'AUZAC, J
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01-02-1994
• Subjects: Biological and medical sciences; Biotechnology; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; Hevea brasiliensis; Methods. Procedures. Technologies; Plant cells and fungal cells; Protoplast preparation, cell fusion and regeneration; Culture medium; Nurse cell; Cell culture; Hevea brasiliensis; Callus; Vegetals; Laticiferous plant; Regeneration; Dicotyledones; Protoplast; Angiospermae; Development; Euphorbiaceae; Kinetin; Plant growth substance; Spermatophyta; Hardwood forest tree
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/BF00233318

Conditions for successful culture of rubber tree (Hevea brasiliensis) protoplasts were investigated. Protoplasts, derived from embryogenic callus, regenerated cell walls then underwent division when embedded in alginate and cultivated on a modified Murashige and Sook medium (9 μM 2,4-dichlorophenoxyacetic acid, 0.6 M glucose, 0.93 μM kinetin, NH 4 (+) reduced by half) in the presence of nurse cells (tobacco feeder cell layer). The presence of nurse cells was essential to maintain viability and sustain protoplast division. Several parameters which influenced the plating efficiency were analysed, such as the density of feeder cells and the duration of contact of the feeder layer.