イネ萎縮病に対する化学療法剤のスクリーニング : I. 技法と応用例
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It has already been reported that rice dwarf virus (RDV) contains double-stranded RNA and that RDV-RNA is easily separated from host cell nucleic acids by methylated albumine-kieselguhr column chromatography and also by cellulose column chromatography. These procedures make it possible to screen such substances which inhibut the incorporation of ^<32>P into RDV-RNA but do not inhibit the ^<32>P-incorporation into ribosomal RNA (rRNA). Two grams of uppermost and second leaves detached from RDV-infected rice plants were placed in a test tube containing 2 ml of aqueous solution of an antibiotic and 60 μc of ^<32>P-H_3PO_4 and kept under greenhouse conditions. After whole the solution was almost absorbed by the leaf sample, 3 ml of water was added to the tube. After 24 hours, nucleic acid was extracted by phenol method any one fourth of the nucleic acid preparation was subjected to cellulose column chromatography. Optical densities at 260 nm of rRNA and RDV-RNA and radioactivities of both RNA's were measured. Inhibitory effect of an antibiotic for the syntheses of both RNA's was evaluated by the decrease of cpm/OD_<260> values. Cycloheximide at 2 μg/g fresh weight inhibited the incorporation of ^<32>P into both rRNA and RDV-RNA equally suggesting that the antibiotic is phytotoxic. This was indeed the case when the antibiotic was applied to RDV-infected rice plants at 2 ppm through culture solution, i. e., the plants died within 2 weeks. Blasticidin S at 3μg/g and kasugamycin at 160 μg/g fresh weight rather stimulated the ^<32>P-incorporation into both rRNA and RDV-RNA, whereas at higher dosages, 6 μg/g and 320μg/g respectively, they inhibited the ^<32>P-incorporation into both RNA's but only very slightly. When these were applied to infected plants through culture solutions at liminal concentrations, i. e., 2 and 40 ppm respectively, throughout the whole life period, there was no sign of the recovery of normal growth.
- 日本植物病理学会の論文
- 1973-01-01
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