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Production of Valuable Metabolites by Plant Hairy Root Cultures and Establishment of System to Harvest the Products

植物毛状根培养物生产有价值的代谢物及产品收获系统的建立

基本信息

批准号：
01550763
负责人：
TAYA Masahito
金额：
$ 1.34万
依托单位：
Osaka 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：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

项目摘要

The induction of hairy roots was tried from various plants and their culture characteristics were investigated in biotechnological a spaects. The following results were obtained.1. By infecting the speciments of carrot, horseradish, puk-bung and red beet with Agrobacteria rhizogenes, the hairy root clones were induced from these plants. Especially, puk-bung hairy root had high activity of peroxidase (valuable as diagnostic enzymes) and its activity was about 5 time that of original plant.2. The growth rates of hairy roots were expressed in terms of branching frequency, elongation rate and decay rate of growing point (meristem) at the tips. It was possible to evaluate the culture properties of hairy roots based on this kinetic growth model. Moreover, the amounts of cell mass and main inorganic nutrients (K^+, NH<_4^+ and NO_3^- could be estimated during the hairy root cultures by the monitoring of medium conductivity.3. The hairy roots from pak-bung and horseradish had high activities of Super-Oxide Dismutase (SOD). A two-step culture of pak-bung hairy root was performed on the basis of the nitrogen source effect on its growth. Namely, medium with NO_3^- was used in cell growth phase and the medium was replaced with NH_4^+ medium in SOD production phase. In this culture system, the SOD productivity was 5-7 times as high as that in the culture with medium containing only NO_3^- as a nitrogen source.4. In the culture of red beet hairy root, significant amounts of intracellular pigments were secreted into medium by the interrupted limitation of oxygen supply. The repeated-batch culture of beet hairy root was carried out with the temporary oxygen starvation, and 20-25 % of the total pigments in cells was recovered from culture broth. Moreover, it was found that styrene-divinylbenzen resin was applicable to the pigment recovery from culture broth as an adsorbent.

从生物技术角度研究了不同植物毛状根的诱导及其培养特性。得到以下结果.用发根农杆菌（Agrobacteriumrhizogenes）感染胡萝卜、山葵、甜菜和红甜菜的种子，诱导出毛状根无性系。特别是北豆根毛状根具有较高的过氧化物酶活性，其活性约为原植物的5倍.毛状根的生长速率用顶端生长点（分生组织）的分枝频率、伸长速率和衰减速率来表示。基于该动力学生长模型可以评价毛状根的培养特性。此外，通过培养基电导率的测定，还可以估算毛状根培养过程中细胞量和主要无机营养盐（K^+、NH_4 ^+和NO_3^-）的含量.白萝卜和辣根毛状根具有较高的超氧化物歧化酶（SOD）活性。根据氮源对其生长的影响，对白棒毛状根进行了两步培养。即在细胞生长期用含NO_3^-的培养基培养，在SOD产生期用含NH_4^+的培养基培养。在此培养体系中，SOD的产量比仅以NO_3^-为氮源的培养体系高5-7倍.在红甜菜毛状根的培养中，通过中断供氧限制，大量的细胞内色素分泌到培养基中。用间歇缺氧法进行甜菜毛状根的重复分批培养，培养液中色素回收率可达20- 25%。苯乙烯-二乙烯基苯树脂作为吸附剂可用于从发酵液中回收色素。