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

试验了多种植物毛状根的诱导，并从生物技术角度研究了毛状根的培养特性。得到了以下结果：用根际农杆菌侵染胡萝卜、辣根、南瓜和红甜菜，诱导出毛状根无性系。其中，pukbung毛状根的过氧化物酶活性较高，是原植物的5倍。毛状根的生长速率以分枝频率、顶端生长点（分生组织）的伸长率和腐烂率来表示。利用该模型可以对毛状根的培养特性进行评价。此外，通过对培养基电导率的监测，还可以估计毛状根培养过程中细胞质量和主要无机营养物质（K^+、NH<_4^+和NO_3^-）的数量。毛根和辣根的超氧化物歧化酶（SOD）活性较高。根据氮源对毛状根生长的影响，对毛状根进行了两步培养。即在细胞生长期用NO_3^-培养基，在SOD产生期用NH_4^+培养基代替。在该培养体系中，SOD的产量是仅含NO_3^-作为氮源培养基的5-7倍。在红甜菜毛状根培养过程中，由于缺氧中断，细胞内大量色素分泌到培养基中。在临时缺氧条件下对甜菜毛状根进行重复分批培养，从培养液中回收细胞总色素的20- 25%。此外，还发现苯乙烯-二乙烯苯树脂作为吸附剂适用于从发酵液中回收色素。

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.