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Development of safer and higher activity herbicides which inhibit acetolactate synthase

开发更安全、活性更高的抑制乙酰乳酸合酶的除草剂

基本信息

批准号：
61470136
负责人：
MATSUNAKA Shooichi
金额：
$ 3.84万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1988
项目状态：
已结题

项目摘要

(1) Selection of plant materials for acetolactate synthase: In spring and early summer, barley, komatsuna, santo-sai and spinach were selected as good materials. In summer, Chinese cabbage and rice plant and in fall spinach and barley were recommended.(2) Extraction and partial purification methods: In a mixture consisting of 0.1 M phosphate buffer(pH 7.0), 1mM pyruvate, 0.5 mM MgCl_2, 0.5 mM TPP, and 10 M FAD, the plant material is homogenized. The precipitate of the homogenate from 20 - 40 % saturated ammonium sulfate-salting out is solved the phosphate buffer, and de-salted using sephdex G-25 column.(3) Assay of the enzyme activity: The mixture of the enzyme, 20mM Na-pyruvate, 0.5 mM NgCl_2 and 0.5 mM TPP is incubated at 30゜C for 30 min.. After the incubation, 6N H_2SO_4 is added to the reaction mixture,which is kept at 60゜C for 15 min..During the process produced acetolactate is decarboxylated into acetoin, which will produce color with the addition of 0.5 % creatin and 5% naphthol and heating again at 60゜C. The optical density of the color is measured at 525 nm.(4) Also effect of chlorsulfuron on the growth of plant and animal tissue cultures, properties of plant acetolactate synthase, the mode of inhibition of chlorsulfuron on ALS, and using 36 reagents and 3 semi-synthesized chemicals evaluation as ALS inhibitors are examined.

(1)乙酰乳酸合成酶植物材料的选择：在春季和初夏，选择大麦、小松菜、山藤菜和菠菜为良好材料。在夏季，建议种植大白菜和水稻，在秋季，建议种植菠菜和大麦。(2)提取和部分纯化方法：在由0.1 M磷酸盐缓冲液（pH 7.0）、1 mM丙酮酸盐、0.5 mM MgCl_2、0.5 mM TPP和10 M FAD组成的混合物中，将植物材料均质化。匀浆经20 - 40%饱和硫酸铵盐析后，用磷酸盐缓冲液溶解沉淀，用Sephadex G-25柱脱盐。(3)酶活性的测定：将酶、20 mM丙酮酸钠、0.5mM NgCl_2和0.5mM TPP的混合物在30 ℃下孵育30分钟。温育后，向反应混合物中加入6 N H_2SO_4，在60 ℃下保持15分钟。在此过程中，产生的乙酰乳酸脱羧为乙偶姻，加入0.5%肌酐和5%萘酚并在60 ℃下再次加热，乙偶姻将产生颜色。在525 nm处测量颜色的光密度。(4)本文还研究了绿磺隆对植物和动物组织培养物生长的影响、植物乙酰乳酸合成酶的性质、绿磺隆对ALS的抑制方式以及用36种试剂和3种半合成化合物作为ALS抑制剂的评价。