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Anthers of Japanese persimmon (Diospyros kaki L. cv. Meotogaki) were cultured on MS medium containing zeatin (0~10 ƒÊM) in combination with IAA, NAA, or 2,4-D (0~10 ƒÊM). Prolific callus formation was induced in the dark on MS medium supplemented with zeatin (1~10 ƒÊM)+ IAA or NAA (1~10 ƒÊM), or MS medium with zeatin 10 ƒÊM +2,4-D (0.001 ~ 0.1 ƒÊM). Those cultured under a 16-hr•Eday-1 photoperiod formed few calli. The calli formed in the dark on MS medium supplemented with 10 ƒÊM zeatin +1 ƒÊM IAA, 0.1 ƒÊM NAA, or 0.1 ƒÊM 2,4-D were subcultured in the dark on the same fresh medium. After 3~5 subcultures, the call were subdivided and transferred onto three different kinds of differentiation medium, MS medium containing a) 10 AM zeatin + 0.1 AM IAA, b) 10 AM zeatin +0.01 ƒÊM NAA, or c) 10 ƒÊM zeatin +0.01 ƒÊM 2,4-D, and cultured under a 16-hr•Eday-1 photoperiod to induce adventitious buds. Organogenetic potential of the calli, which had been induced and subcultured on MS medium containing IAA or NAA, increased with increasing number of subcultures, whereas those which had been cultured in the presence of 2,4-D formed few adventitious buds. IAA or NAA in the differentiation medium was also more effective for adventitious bud formation than was 2,4-D. Both histological observations of the development of anther callus and chromosome counts of the root tip cells of regenerated plantlets revealed that the calli originated not from pollen grains but from vascular bundle in the anthers. Eight of 18 plantlets regenerated from the anther-derived calli showed different malate dehydrogenase isozyme banding patterns from the pattern of the 'Meotogaki' tree growing in the field. However, all 12 plantlets regenerated from mill derived from leaf primordia of dormant buds of 'Meotogaki', had the same banding patterns as 'Meotogaki'. The potential use of the somaclonal variation observed in the regenerated plantlets from anther-derived calli is discussed in relation to the cultivar improvement of Japanese persimmon.

Adventitious Bud Formation on Callus Derived form Anthers of Persimmon 'Meotogaki' and Isozyme Variations Observed in the Regenerated Plantlets.