| Budget Amount *help |
¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1988: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Research Abstract |
(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.
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