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Plants were regenerated from protoplasts isolated from hypocotyls of Iris japonica cv. 'Tsugaru'. With an initial rate of protoplast formation of 50% or higher, the transformation ratio was 100%. Shoot regeneration without callus formation was attained on MS medium containing 5 mg/l naphthaleneacetic acid. Shoot regeneration from protoplasts was possible only in the presence of naphthaleneacetic acid. In the regeneration medium containing 1-10 mg/l naphthaleneacetic acid, the highest percentage of shoots was attained with 5 mg/l (78%). Shoot regeneration from protoplasts was more frequent when the hypocotyls were older. The number of regenerated plants was the same whether the protoplasts were transferred to the regeneration medium containing 5 mg/l naphthaleneacetic acid from the protoplast isolation medium or directly transferred from the isolation medium. Morphologically, the regenerants could be divided into two types, small (i.e., about 1 mm) and large (about 10 mm) plants. Shoot elongation was the highest from 6th to 13th days after protoplast isolation, and then shoot growth was retarded. Within a regenerant population, however, the number of large plants was much larger than the number of small plants. The embryogenic nature of isolated protoplasts was considered by the presence of embryoid masses and subsequent shoot regenerations from protoplasts.Routine screening of 0b46394aab