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An enhanced embryo culture methodology for coconut (Cocos nucifera l.)

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Pablito M. Magdalita, Olivia P. Damasco, Yohannes M.S. Samosir, Nurhaini Mashud, Hengky Novariento, Erlinda P. Rillo, Vu Thi My Lien, Alfred Kembu, Mathias Fauare and Stephen W. Adkins

Improvements to the conventional embryo culture protocol have been initiated. These improvements, among several, include the use of a CO2-enrichment step within a photoautotrophic culture system and the  application of 100 µM of either α-napthalene acetic acid (NAA) or indolebutyric acid (IBA) to promote rooting of seedling. To obtain uniform germination of embryos of different ages, abscisic acid (ABA) at 0.97-7.67 µM is added to the germination medium.  ABA could be replaced with gibberelic acid (GA3) at 10.00 µM when handling varieties that are difficult to germinate.  Following the in vitro acclimatization using the CO2 enrichment system, the hardening-off process ex vitro in the screenhouse is done by placing the seedlings inside the acclimatization box that provides high relative humidity and sufficient headspace for growth.  Alternatively, a clear plastic covered tent could be used instead when dealing with large numbers of seedlings.  By using this enhanced methodology on the embryos of normal coconuts the in vitro stage can now be reduced from 10 to 12, to 3 to 4 months and the success rate in transferring embryos to seedlings in the field was improved from 50 to 100%. The enhanced methodology needs to be further refined and applied to the different mutant coconut types that are found around the world.

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