ラットチトクロームP4501A1を発現したトランスジェニックバレイショの除草剤代謝と耐性

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  • Herbicide Metabolism and Resistance of Transgenic Potato Plants Expressing Rat Cytochrome P4501A1.
  • Herbicide metabolism and resistance of

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It was attempted to generate transgenic potato plants expressing rat P4501A1 and rat P4501A1/yeast reductase (YR) fused enzyme by the use of Agrobacterium-transformation system. Six regenerated plants for P4501A1 (GC)and twenty plants for P4501A1/YR fused enzyme (GFC) were selected by kanamycin resistance. Southern blot analysis revealed that one of GC plants and ten of GFC plants showed several bands of P450 cDNA or its fused enzyme gene. The GC plant N0.1160 was found to produce a detectable amount of mRNA corresponding to 1.6 kb-P450 cDNA, whereas the GFC plants gave a much less amount of mRNA corresponding to 3.5 kb-P450/YR fused enzyme gene. Western blot analysis showed that the GC plant N0.1160 produced a large amount of a 59kDa protein corresponding to rat P4501A1. No GFC plants gave a detectable amount of a 130kDa fused protein corresponding to rat P4501A1/YR fused enzyme. 7-Ethoxycoumarin O-deethylase and cytochrome c oxidoreductase activities of the transgenic plants were 1.4 to 3.5 and 1.3 to 3.5 times higher than those of the control plants, respectively. [ˆ14C]-Labeled chlortoluron(CT) was more rapidly metabolized through N-demethylation and p-methyl hydroxylation to none or less phytotoxic metabolites in the GC plant N0.1160, while it was also metabolized through N-demethylation in the control plants. In vivo herbicide-tolerant tests revealed that the GC plant N0.1160 was tolerant against the spray of the CT at 10μmol per pot and another herbicide DCMU, [3-(3, 4-dichlorophenyl)-1, 1-dimethylurea], at 2μmol per pot, although the control plant completely died under the same concentrations of both herbicides. Thus, it was found that expression of rat P4501A1 cDNA confer tolerance to the phenylurea herbicides in potato plants.

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