DEVELOPMENT OF PRODUCTION PROCESS OF ARTIFICIAL SEEDS USING CELL AGGREGATES DERIVED FROM PLANT HAIRY ROOTS

利用植物毛状根细胞聚集体开发人工种子生产工艺

• 批准号: 06555246
• 负责人: TONE Setsuji
• 金额: $ 3.97万
• 依托单位: Osaka 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: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06555246/
• 关键词: BIOCHEMICAL ENGINEERING; ARTIFICIAL SEEDS; LONG-TERM PRESERVATION; HORSERADISH HAIRY ROOTS; TOTIPOTENCY; PLANT CELL CULTURE; MASS PROPAGATION; ARTIFICIAL EMBRYOS; 育種; 植物毛状根; 形態変化

The aim of the present research is to develop artificial seeds for the mass propagation of useful plant variants. This research is comprised of the following parts : 1) process of cell aggregate (referred to as CA) production from hairy roots, 2) preparation of artificial seeds by CA encapsulation, and 3) process of root emergence and plant development from artificial seeds. In each process, the hairy root cells were used because of their genetic stability and fast growth rate, and the following results were obtained :1. The medium with phytohormones, namely, BAP (6-benzylaminopurine) and NAA (1-naphthaleneacetic acid) was effective for the formation of CA from horseradish hairy roots. It was demonstrated that these CA were useful as artificial embryos. The synergistic effects of BAP and NAA on the CA formation were also clarified with a kinetic model.2. During encapsulation of CA in alginate gel beads, it was shown that the regenetation potential of CA could be enhanced by NAA treatment. To improve the storage potential of the artificial seeds, the application of coats on the alginate gel surface was considered, and paraffin was found to be the most suitable among the various materials tested.3. It was observed that the root emergence from CA could be expressed as a normal distribution function of time, and the kinetics of CA regeneration was analyzed. Moreover, it was shown that the regenerating CA subsequently developed into whole plants upon transfer under light condition. On the other hand, the relation between regeneration effeciency and permeability of paraffin coat membrane to oxygen was elucidated, and the regeneration potential of CAA after 60 days of storage was verified.It is considered that the above findings could contribute to the advancement of artificial seed technology for the preservation and mass propagation of useful plants.

本研究的目的是开发人工种子用于大量繁殖有用的植物变体。本研究包括以下几个部分：1）毛状根细胞团（简称CA）的产生过程; 2）CA包埋人工种子的制备; 3）人工种子的根萌发和植株发育过程。由于毛状根细胞具有遗传稳定性好、生长速度快等特点，在各工艺中均采用了毛状根细胞，并取得了以下结果：1。在含有植物激素BAP（6-苄氨基嘌呤）和NAA（1-萘乙酸）的培养基上，辣根毛状根CA的形成是有效的。结果表明，这些CA是有用的人工胚胎。通过动力学模型研究了BAP和NAA对CA形成的协同作用.在海藻酸钠凝胶微球包埋CA的过程中，NAA处理可提高CA的再生能力。为了提高人工种子的贮藏能力，在海藻酸盐凝胶表面进行包衣处理，发现石蜡是最合适的包衣材料.结果表明，CA根的出现可以用时间的正态分布函数表示，并对CA再生动力学进行了分析。此外，研究表明，再生CA随后在光照条件下转移后发育成完整植株。另外，还阐明了再生效率与石蜡包被膜透氧性的关系，并验证了CAA保存60天后的再生潜力，认为上述研究结果有助于促进有用植物保存和大量繁殖的人工种子技术。

项目成果

Vez P.Repunte: "化学工学シンポジウムシリーズ37「植物機能および植物生産物利用技術の新展開」" 化学工学会, 150 (1994)

Vez P.Repunte：“化学工程研讨会系列 37 “植物功能和利用植物产品的技术的新发展””化学工程师学会，150 (1994)

Vez P.Repunte, Shigetaka Shimamura, Masahito Taya and Setsuji Tone: "Kinetic Analysis of the Process of Hormone-Induced Cell Aggregate Formation from Horseradish Hairy Roots" J.Chem.Eng.Japan. 28. 847-850 (1995)

Vez P.Repunte、Shigetaka Shimamura、Masahito Taya 和 Setsuji Tone：“辣根毛状根激素诱导细胞聚集体形成过程的动力学分析”J.Chem.Eng.Japan。

嶋村重孝: "化学工学シンポジウムシリーズ37「植物機能および植物生産物利用技術の新展開」" 化学工学会, 150 (1994)

Shigetaka Shimamura：“化学工程研讨会系列 37 ``植物功能和植物生产利用技术的新发展”化学工程师协会，150（1994）

Vez P.Repunte: "セイヨウワサビ毛状根から誘導された細胞塊の保存活性に及ぼす環境因子の影響" 化学工学会第60年会講演要旨集. 3. 136 (1995)

Vez P. Repunte：“环境因素对辣根毛状根细胞团保存活性的影响”第 60 届化学工程师学会年会记录（1995 年）。

Vez P.Repunte: "パラフィンで被膜された人工種子からの発芽効率に及ぼす保存条件の効果" 化学工学会第61年次講演要旨集. 2. 9 (1996)

Vez P.Repunte：“储存条件对石蜡包被的人造种子发芽效率的影响”化学工程师学会第 61 届年会记录（1996 年）。