GENETIC-TRANSFORMATION OF JAPANESE PERSIMMON (DIOSPYROS-KAKI L) BY AGROBACTERIUM-RHIZOGENES WILD-TYPE STRAIN A4

Crown gall tumors were obtained from three cultivars, 'Fuyu', 'Jiro', and 'Nishimurawase', of Japanese persimmon incited with A. rhizogenes wild type strain A4 harboring agropine type Ri plasmid (pRiA4b). When the tumors were cultured on MS (1/2N) medium containing 10 muM zeatin, 10 muM IAA, and 500 mg.l-1 cefotaxime, calli were induced from the tumors. After the establishment of axemic cultures, more than 60% of the tumor-derived calli formed adventitious buds on MS medium containing 10 muM zeatin and 0.1 muM IAA. These buds could be developed into shoots on MS medium supplemented with 5 muM zeatin and 1 muM IBA. PCR and Southern blot analyses revealed that some of the regenerants from tumor-derived calli were Ri TL-DNA transformants but that non-transformants were regenerated also from the tumor-derived calli. Phenotypic alterations such as dwarfness and decrease in rooting ability were observed in the transformants.