Adventitious Bud Formation on Callus Derived from Anthers of Persimmon 'Meotogaki' and Isozyme Variations Observed in the Regenerated Plantlets
1992 Volume 61 Issue 3 Pages 527-533
Details
1992 Volume 61 Issue 3 Pages 527-533
Anthers of Japanese persimmon (Diospyros kaki L. cv. Meotogaki) were cultured on MS medium containing zeatin (010 μM) in combination with IAA, NAA, or 2, 4-D (010 μM).
Prolific callus formation was induced in the dark on MS medium supplemented with zea-tin (110 μM) + IAA or NAA (110 μM), or MS medium with zeatin 10 μM +2, 4-D (0.0010.1 μM). Those cultured under a 16-hr•day-1 photoperiod formed few calli.
The calli formed in the dark on MS medium supplemented with 10 μM zeatin +1 μM IAA, 0.1 μM NAA, or 0.1 μM 2, 4-D were subcultured in the dark on the same fresh medium. After 35 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 μM NAA, or c) 10 μM zeatin +0.01 μM 2, 4-D, and cultured under a 16-hr•day-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 de-hydrogenase 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.
