Development of artificial seed using plant hairy root

利用植物毛状根开发人工种子

• 批准号: 03556012
• 负责人: KOBAYASHI Takeshi
• 金额: $ 4.93万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03556012/
• 关键词: artificial seed; hairy root; regeneration; horseradish; adventitious shoot; Agrobacterium rhizogenes; phytohormone; adventitious shoot primordium; オ-キシン; 大量培養

We focused on the development of micropropagation procedure of horseradish hairy root by using artificial seed system from the standpoint of bioengineering. 1. Auxin supplementation stimulated branching, which resulted in the increment of the number of apical meristem and cell growth rate. The kinetic growth model was developed to simulate the hairy root growth. 2. The root fragments with branch or apical meristem excised by blade were highly regenerated under dark conditions. 3 The adventitious shoot primordium on the horseradish hairy root by microscopic morphological observation and alteration to plantlet at the exposure to light is identified. The adventitious shoot primordium offers an advantage for relatively higher synchrony in development at exposure to light, compared with two other cells. It is necessary to increase the number of the adventitious shoot primordium formation in an artificial seed system. 4. Plantlet developed from hairy root were obtained by treatment cytokinin supplementation. To cut the root into fragments, the blender was used. The mechanical excision improved the bioprocess to produce the competent root fragments as artificial seed. the bioreactor, the regeneration frequency was similar to that of that in shake flask. The candidate hairy root cells included in beads were classified into three ; 1, the root fragment cultured under dark condition. 2, the adventitious shoot primordium formed under dark condition. 3, plantlet developed from the hairy root which was formed after transferred into light. The above results show the possibility of application of hairy root for artificial seed production.

从生物工程的角度出发，研究了辣根毛状根的人工种子快繁技术。1.生长素的添加促进了分枝的形成，从而导致顶端分生组织数目的增加和细胞生长速率的提高。建立了毛状根生长的动力学模型。2.用叶片切下带有分支或顶端分生组织的根片段，在黑暗条件下再生率较高。3.通过显微形态观察和光照对植株的影响，对辣根毛状根上的不定芽原基进行了鉴定。与其他两种细胞相比，不定芽原基在光照下的发育具有相对较高的同步性。在人工种子系统中，增加不定芽原基的形成是十分必要的。4.用细胞分裂素处理毛状根，获得毛状根分化植株。为了把根切成碎片，使用了搅拌机。机械切除改善了生物过程，产生了作为人工种子的感受态根片段。在生物反应器中，再生频率与摇瓶相似。珠子中包含的候选毛状根细胞分为三种：1，在黑暗条件下培养的根片段。2、黑暗条件下不定芽原基形成。3.转光后形成的毛状根发育成植株。上述结果表明，毛状根在人工种子生产中应用的可能性。

项目成果

N.Uozumi: "Fed-batch culture of hairy root cells using fructose as a carbon source" J.Ferment.Bioeng.72. 457-460 (1991)

N.Uozumi：“使用果糖作为碳源的毛根细胞补料分批培养”J.Ferment.Bioeng.72。

N.Uozumi: "Production of artificial seed from horseradish hairy root" J.Ferment.Bioeng.

N.Uozumi：“用辣根毛状根生产人工种子”J.Ferment.Bioeng。

Nobuyuki UOZUMI: "Production of artificial seed from horseradish hairy root" Journal of Fermentation and Bioengineering. 74. 21-26 (1992)

Nobuyuki UOZUMI：“用辣根毛状根生产人造种子”《发酵与生物工程杂志》。