Micropropagation protocol for pineapple

A technique to rapidly multiply promising hybrids and clones of pineapple, which can serve as back-up copies of the hybrids in culture was developed. Though pineapple can be easily propagated asexually through slips, suckers, and crowns, the rate of increase over time is slow. This is especially true where the starting mother plant is a hybrid derived from a single plant selection. The micropropagation protocol and the media used are as follows: A Protocol: Explant preparation-wash the healthy suckers or slips with running water to remove soil particles; cut the basal portion of the explant to facilitate leaf removal; trim the leaf and the base to about 7.6 cm long; Explant sterilization-place the explant in large 2L capacity beakers and wash with soap and water; Add two to three drops of Tween 20.; surface sterilize explant with 10% commercial calcium hypochlorite or sodium hypochlorite solution for 20 minutes; rinse three times with sterile distilled water. Culture procedure-dissect the explant inside a sterile laminar flow hood by trimming the basal portion and removing excess leaves. Leave only 1 cm of the base and 2-3 leaves; Culture the dissected explant in shoot induction medium; Subculture every three weeks; Place the culture in shoot regulation media when the shoot has produced a new set of leaves; subculture every three weeks; When the shoot has produced 10-15 leaves, divide the plantlet vertically into two sections and place in liquid proliferation media. Subculture every two weeks; Separate new plantlets having at least five leaves from proliferating culture and place in shoot regulation media. Subculture every three weeks; Proliferate or root the plantlet and then pot out in single pots. Place the plantlets in an elevated platform inside the screenhouse; Acclimatize the culture while they are still inside the vessel. Loosen the vessel's lid in order to expose the plantlet gradually to room temperature for two days. Remove the plantlet from the vessel, plant in sterile potting medium, and then place in the mist bed. If the mist bed is not available, use an inverted bottle to protect the young plantlet. Remove the inverted bottle after two days