MOLECULAR BIOLOGICAL AND ENBIRONMENT CHEMICAL APPROACHES FOR GERMINATION CONTROL OF RED-TIDE ALGEA

目前的研究资料表明，除了工业污染造成的高浓度营养因子外，赤潮藻囊萌发的波动可能是引发赤潮的主要因素。为寻找和开发控制赤潮藻包囊萌发的方法，进行了三年的分子生物学和环境化学研究。对濑户内海Harima Nada地区查顿菌囊分布谱的研究表明，从约1900年开始，这一百年来柱形沉淀物中查顿菌囊数量呈上升趋势，在1960-1970年达到峰值后迅速下降。数据表明，公元1600年，Harima Nada已经出现了红潮藻类，chatonella sp.。对不同处理后的包囊细胞的潜在萌发能力进行了分析。对包囊细胞施加100V，5min的电流消除了包囊的萌发能力。结果表明，该技术在控制包囊萌发方面具有较好的应用前景。制备了5种抗恙虫病斯克里普菌包囊的单克隆抗体，并对其反应性进行了观察。数据显示抗体只对囊肿的棘起反应。对营养性生长藻类无反应性。底土ELISA法成功测定了土壤中包囊的含量。用8 ~ 12分子量的singer引物对查顿菌、亚历山大菌、斯克里普菌和异源菌进行了DNA指纹图谱的测定。银染指纹图谱显示种内或种内特异性条带，可用于田间赤潮藻的鉴定。分离纯化了Gymunodium A3和Cocholodium polykrikoides的酸性多糖，并对其结构进行了研究。经电泳和分析性超离心鉴定，纯化后的部分为均质。它由焦糖、甘露糖、半乳糖、葡萄糖和半乳糖醛酸组成。多糖对培养细胞无明显的毒性作用。少

Current research data suggest that fluctuation of cyst germination of red-tide algae can be a major factor to start red tide, in addition to high concentration of nutritional factors by industrial polution. To find and develop the methods to control the germination of cysts of the red-tide algae, molecular biological and environment chemical studies were continued for three years.The study on profiles of cyst (Chattonella sp.) distribution in Harima Nada in Seto Inland Sea revieled that cyst number in cylidorial precipitate during these hundred years increased from ca.1900AD,and the number decreased rapidly then after a peak in 1960-1970. The data demonstrated that red-tide algae, Chattonella sp., was already present at Harima Nada in 1600AD.The potential ability of germination of the cyst cells were analyzed after various treatments. Electronic current application (100V,5min) to the cyst cells eliminated the ability of germination of the cysts. The data indicated the technique was use … More full for cyst germination control.Five kinds of monoclonal antibodies against cysts of Scrippsiella trochoidea were raised and the reactivity was surveyed. The data showed that the antibodies were reacted only against spines of the cysts. No reactivity was detected against nutritional growing algae. ELISA of the bottom soil successfully determined the contents of the cysts in the soil.DNA fingerprintings of Chattonella sp., Alexandrium sp., Scrippsiella sp.and Heterosigma sp.were surveyed with singer primers of 8 mer to 12 mer. The finger print visualized by silver stain showed species specific or intraspecies specific band patterns, and the technique can be adaptable to identify red tide algae in the fields.Acidic polysaccharides from Gymunodium A3 and Cocholodium polykrikoides were purified and the structures were surveyed. The purified fraction was identified to be homogenous by electrophoresis and by analytical ultracentrifugation. It was consisted of fucose, mannose, galactose, glucose and galacturonic acid. The polysaccharide did not show clear toxic effect to cultured cells. Less