Production of intergeneric hybrids of wasabi and horseradish by somatic cell fusion.

通过体细胞融合生产芥末和辣根的属间杂种。

• 批准号: 04660025
• 负责人: OHI Michio
• 金额: $ 1.54万
• 依托单位: Shinshu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04660025/
• 关键词: Wasabi; Horseradish; Electric cell fusion; Asymmetrical hybridization; Beads culture; ビーズ培養; プロトプラスト; サイトプラスト; アルギン酸; FDA染色; 細胞融合; PEG法; 電気融合法

Mature leaves of both species obtained from the plants in vitro were used for this experiment. Percentage of formation of cell fusion and single pair was raised 17% and 8% by increase of pulse number, respectively.Removal of nucleus for asymmetrical hybridization was carried out by using Percoll. The vigorous miniprotoplast and cytoplast were obtained. However, the protoplasts which were not removed the nuclei remained, and subprotoplast was litte obtained. reciprocal fusion of protoplast and miniprotoplast, miniprotoplast and cytoplast, and protoplast and cytoplast was carried out. Cytoplasm of one parental cell was introduced smoothly another parental cell by the fusion of protoplast and cytoplast. In contrast, Introsuction and substitution of nuclear was difficult by electric cell fusion method. The beads of alginic acid gel which contained the cells substituted cytoplasm were cultured. The beads culture was suitable for this experimental purpose comparing with liquid culture. We are, presently, attempting search of culture conditions induced the formation of colony.

本实验采用两种植物离体的成熟叶片。增加脉冲数可使细胞融合率提高17%，单双对率提高8%。用Percoll法去除细胞核进行不对称杂交。获得了充满活力的小原生质体和细胞质。而未除去细胞核的原生质体保留了下来，亚原生质体很少得到。原生质体与小原生质体、小原生质体与细胞质、原生质体与细胞质相互融合。原生质体与细胞质的融合使一个亲本细胞的细胞质顺利引入另一个亲本细胞。相比之下，电细胞融合法很难实现核的内吸和取代。培养含有细胞的海藻酸凝胶珠代替细胞质。与液体培养相比，微球培养更适合于实验目的。目前，我们正试图寻找诱导群体形成的培养条件。