Embryo culture of coconut: the CPCRI protocol

The field collection of embryos from mature nuts (9 months onwards), which can be easily identified for its hard eye and thick kernel, is done by means of a surface-sterilized cork borer. From the endosperm plug thus scooped out, the embryo is separated by using knife. The collected embryos are placed in distilled water. Once all the embryos are extracted, they are subjected to surface sterilization with 50% chlorine water for 20 minutes. The embryos are washed thoroughly in sterile water 4–5 times and inoculated individually into screw-cap bottles containing 2–5 ml sterile water (pH 5.7). The entire operation is to be done in surface-sterilized (absolute alcohol) inoculation hood (portable). The field-collected embryos are then transferred to Y3 solid medium within 2 months. The medium is supplemented with 30 g/l sucrose and 1 g/l charcoal for embryos of tall types and 60 g/l sucrose and 2.5 g/l charcoal for dwarf types.