Department of Biotechnological Science, Faculty of Biology Oriented Science and Technology, Kinki University, Uchita, 649-64, Wakayama, Japan.
Plant Cell Rep. 1996 Mar;15(7):470-3. doi: 10.1007/BF00232976.
Dodecaploid plants of Japanese persimmon (Diospyros kaki L.) were obtained by colchicine treatment of protoplasts. Callus protoplasts of 'Jiro' (2n=90, x=15) were cultured in modified KM8p medium with 0.1% colchicine for 3-9 days. After colchicine treatment, they were cultured using agarose bead culture. Microcalli were recovered from the protoplasts after 3 months. Flow cytometric measurement showed that nine of 31 callus lines obtained from 6 days of colchicine treatment had twice the nuclear DNA content as non-treated controls. Plantlets were regenerated from the calli with twice the nuclear DNA content. Microscopic observation of root tip cells showed that their somatic chromosome number was 2n=180 (x=15). Compared with 'Jiro', dodecaploid plants had longer stomatal guard cells and lower stomatal densities, consistent with increased ploidy.
日本柿子（Diospyros kaki L.）的十二倍体植物是通过原生质体秋水仙素处理获得的。“次郎”（2n=90，x=15）的愈伤组织原生质体在改良的 KM8p 培养基中用 0.1%秋水仙素培养 3-9 天。秋水仙素处理后，采用琼脂珠培养进行培养。3 个月后从原生质体中回收微愈伤组织。流式细胞术测量表明，在 6 天的秋水仙素处理中获得的 31 个愈伤组织系中有 9 个的核 DNA 含量是未处理对照的两倍。具有两倍核 DNA 含量的愈伤组织再生出植株。根尖细胞的显微镜观察表明，其体细胞染色体数为 2n=180（x=15）。与“次郎”相比，十二倍体植物的气孔保卫细胞更长，气孔密度更低，这与倍性增加一致。
