Journal
PLANT SCIENCE
Volume 210, Issue -, Pages 118-127Publisher
ELSEVIER IRELAND LTD
DOI: 10.1016/j.plantsci.2013.05.021
Keywords
Oil palm; Suspension cultures; Protoplasts; Plant regeneration
Categories
Funding
- MPOB's Board approved collaborative project Establishment of Transgenic Oil Palm with High Added Value for commercial exploitation [T0003050000-RB01-J]
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Oil palm protoplasts are suitable as a starting material for the production of oil palm plants with new traits using approaches such as somatic hybridization, but attempts to regenerate viable plants from protoplasts have failed thus far. Here we demonstrate, for the first time, the regeneration of viable plants from protoplasts isolated from cell suspension cultures. We achieved a protoplast yield of 1.14 x 10(6) per gram fresh weight with a viability of 82% by incubating the callus in a digestion solution comprising 2% cellulase, 1% pectinase, 0.5% cellulase onuzuka R10, 0.1% pectolyase Y23, 3% KCl, 0.5% CaCl2 and 3.6% mannitol. The regeneration of protoplasts into viable plants required media optimization, the inclusion of plant growth regulators and the correct culture technique. Microcalli derived from protoplasts were obtained by establishing agarose bead cultures using Y3A medium supplemented with 10 mu M naphthalene acetic acid, 2 mu M 2,4-dichlorophenoxyacetic acid, 2 mu M indole-3-butyric acid, 2 mu M gibberellic acid and 2 mu M 2-gamma-dimethylallylaminopurine. Small plantlets were regenerated from microcalli by somatic embryogenesis after successive subculturing steps in medium with limiting amounts of growth regulators supplemented with 200 mg/l ascorbic acid. (C) 2013 Elsevier Ireland Ltd. All rights reserved.
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