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Makapuno and the Embryo Culture Technology

Makapuno, a rich ingredient for ice creams, pastries, and sweet preserves, is a mutant coconut where, instead of coconut water, it contains a soft white jelly-like meat that almost fills the nut. This is why it is called makapuno; in Tagalog, it means “to fill”. Traditional coconut palms give only about 10-15% makapuno nuts and their occurrences appear to be a chance event only. Although the makapuno nut contains a normal embryo, it fails to develop because of the possibility that as the nut matures, the meat fills the nut; or that it lacks the milky fluid; or that the over-growing embryo lacks the sufficient nutrients which it needs, that eventually the nut dies making it incapable of reproducing itself. When food manufacturers increasingly demanded for more makapuno, the problem for its reproduction came about. Fortunately, in the early sixties, a female botanist came to its rescue. Born on September 22, 1929 in Binmaley, Pangasinan, Dr. Emerita V. de Guzman (19291981) was a sickly yet an intelligent student. She graduated from elementary and high school as class valedictorian. She was said to be inclined to botany and field work. She acquired her degree in Botany in 1953 as magna cum laude at the University of the Philippines in Diliman. She took a teaching job at the University of the Philippines in Los Baños (UPLB) until she was sent to Cornell University to pursue her masters and doctoral degrees in plant physiology. Knowing that producing makapuno is only a chance event, Dr. de Guzman was convinced that she can do something to improve its low percentage and grow a tree that will produce 100% makapuno nuts. She took the challenge of saving the makapuno embryo and successfully isolated and grew it to be a true makapunobreeding tree. This started the production of embryo cultured makapuno (ECM) which used the embryo culture technology.

The embryo culture technology refers to the artificial growing of the embryo by supplying it with essential nutrients as well as placing it in favorable conditions for its growth and development into a seedling that may successfully be transplanted on the field later on. The University of the Philippines, supported by the National Research Council of the Philippines, initiated the development of the makapuno embryo culture to solve the problem on makapuno’s inability to reproduce. The makapuno embryo culture technology can be simplified into eight steps, namely; harvesting, dehusking, splitting of the nuts, extraction of the endosperm, sterilization, media preparation, decontamination and the transferring of the culture to screenhouses. To start the makapuno embryo culture, 10 to 11 months old coconuts are harvested. Younger coconuts do not respond very well with the conditions of embryo culture. The nuts are then dehusked and splitted into halves by striking the longitudinal vein that runs around the nut. The embryo enclosed in the solid meat called the ‘endosperm’ is located under one of the three “eyes” of the coconut. The inability of this eye to harden, or its non-lignification cells makes it soft, thereby, allowing the embryos easier fertilization. Using a cork borer, the embryos are then extracted. The endosperm cylinders, the ones left after the embryos are removed, are pushed out of the cork borer using a piece of stick. These are placed in a clean container with coconut or plain water. Inside the laboratory, the endosperm cores that were once surrounding the embryo, which have been extracted, are decontaminated with commercial bleach and rinsed with sterile water. Under sterile condition, the embryos that were removed from the endosperm are planted into a prepared medium. The cultures are then incubated under light condition.

The cultures go through a series of extreme exclusion from microorganisms that might contaminate them. This is known as the aseptic technique. Instruments being used in the culture as well as the plant material itself must be sterile. To reduce the risk of contamination, cleanliness, efficient organization and routine sterilization of all materials are observed. Finally, those seedlings that have at least one primary root and one expanded leaf are ready for soil establishment. From the laboratory, they are brought out so that they may adapt to natural light and temperature conditions. They are placed in a screenhouse for three to five days while still inside their culture vessels. They are then taken out from these vessels, rinsed with water, and dipped in fungicide solution. They are planted to soil medium containing one part garden soil and one part coconut coir dust and incubated inside a humidity tent. After at least one month, the established seedlings are transferred to another screenhouse for hardening. Hardened seedlings are transferred to a nursery for further growth and development until ready to be planted on the field. Indeed, what overturned the makapuno production in the Philippines is the success of the makapuno embryo culture technique where Dr. de Guzman painstakingly developed. However, the optimization of this technique for commercial application is through the initiative of Mrs. Erlinda P. Rillo of the Philippine Coconut Authority (PCA) Albay Research Center. Now, pure makapuno seedlings are made affordable to all – food manufacturers and Filipino coconut farmers alike. (Pauline Mae P. Araneta)