Molecular ecology for the purpose of terminating toxic dinoflagellate by gene analysis of some enzymes relating to PSP toxin synethesis.

分子生态学，目的是通过对一些与 PSP 毒素合成相关的酶进行基因分析来终止有毒甲藻。

基本信息

批准号：
04404014
负责人：
ISHIDA Yuzaburo
金额：
$ 18.56万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1994
项目状态：
已结题

项目摘要

We investigated the stability of paralytic shellfish poisoning (PSP) toxin composition in the dinoflagellate Alexandrium catenella and A.tamarense, and the genetic system producing it. Using clonal axnic cultures, toxin compositions were determined in cell material from different growth phases of many populations and F_1progenies. In all isolates used, the mole percentage of toxin composition did not significantly change in any growth phase. Toxin composition analysis of over 70 strains of both species from 8 different waters indicate the occurrence of inter-and intra-specific indigenous populations from distant localities. Toxin compositions of many F_1progenies from crosses between algal strains having different toxin profiles were determined. Toxin compositions were inherited mostly in a 1 : 1 Mendelian pattern and sexuality (mating type) was not associated with toxin inheritance. These data strongly suggest that PSP toxin compositions of A.catenella and A.tamarense are stable and that Mendelian inheritance of toxin profiles occurs in both these heterothallic dinoflagellates.Furthermore we have tried to isolate some enzymes relating to PSP biosynthesis in A.tamarense and G.catenatum. Firstly, N-sulfotransferase from STX or GTX2+3 to GTX5 or C1+2 was purified from G.catenatum through several columns. The enzyme consisted of two identical subunits (63kDa) and was optmally active at pH6.0 and 25ﾟC.The activity was stimulated by Mg^<2+> and inhibited by PAPS analogue. Secondly, O-sulfotransferase which converted 11-alpha, beta -hydroxy STX into GTX2+3 was partially purified. DNA sequence of these enzymes will be analyzed in the coming year.

我们研究了麻痹贝类中毒（PSP）毒素组成在鞭毛鞭毛脂蛋白Alexandrium catenella和a.tamarense中的稳定性，以及产生它的遗传系统。利用克隆轴向培养物，在许多种群和F_1progenies的不同生长阶段的细胞材料中确定毒素成分。在所有使用的分离株中，毒素成分的摩尔百分比在任何生长阶段都没有显着变化。对来自8种不同水域的两种物种的70种菌株的毒素组成分析表明，遥远地区的特异性本地种群发生。从具有不同毒素谱的藻类菌株之间的杂交中的许多F_1肌基因组成。毒素成分主要以1：1的孟德尔模式和性（交配类型）与毒素遗传无关。 These data strongly suggest that PSP toxin compositions of A.catenella and A.tamarense are stable and that Mendelian inheritance of toxin profiles occurs in both these heterothallic dinoflagellates.Furthermore we have tried to isolate some enzymes relating to PSP biosynthesis in A.tamarense and G.catenatum.首先，通过几列从G.catenatum纯化了从STX或GTX2+3到GTX5或C1+2的N-硫代转移酶。该酶由两个相同的亚基（63KDA）组成，在PH6.0和25°C上具有最佳活性C。其次，将11-α，β-羟基STX转换为GTX2+3的O-硫代转移酶被部分纯化。这些酶的DNA序列将在来年进行分析。