In situ regeneration of rice (Oryza sativa L.) callus immobilized in polyurethane foam

An in situ regeneration system for rice calli comprised of a callus growth stage and two regeneration stages was developed. After the first stage of regeneration, the medium is changed and the calli are immobilized in polyurethane foam supports, in each of which 3-5 regenerated plantlets develop from the immobilized calli during the second stage. While no significant change in callus size was observed during the first stage of regeneration, in the second stage callus enlargement and shoot regeneration predominated. In the light of these findings, calli were immobilized in the second stage after medium exchange. The use of 10-mm support cubes with an average pore size of 3.6 mm resulted in the most efficient immobilization and in situ regeneration. Medium exchange after 15 d gave the largest number of support cubes with shoots. When rice calli were cultivated in support cubes placed in 60 ml second-stage medium in a 500-ml flask, the immobilization ratio was 83%, and 82% of the support cubes contained 3-5 regenerated plantlets after 25 d. The shoot lengths of the regenerated plantlets obtained from the in situ regeneration culture were longer than those from a suspension culture. When support cubes with 3-5 regenerated plantlets were transferred from the flask to 1 4 MS solid medium supplemented with 10 g/l sorbitol and 5 g/l sucrose, the regenerated plantlets developed quickly into plants with a length above 10 cm after 10 d.