D-44 YZI-1S細胞におけるProtoporphyrinogen oxidase阻害型除草剤抵抗性の分子解析

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Protoporphyrinogen (Protox) inhibiting herbicides specifically inhibit Protox which is the last enzyme of the common branch of the chlorophyll and heme synthesis pathway. We have previously selected S23142 resistant line of photomixotrophic tobacco cells, YZI-1S, by stepwise increase of the herbicide, and proporsed two different mechanisms of resistance to S23142: overproduction of the target enzyme and a change of Protox sensitivitiy to the herbicides by some mutation of the Protox encoding gene. In this study we report the sequences and expression levels of plastid and mitochondrial Protox in wild type and YZI-1S cells as well as morphological characters of these cells. The sequence analysis of the cloned Protox cDNAs showed that the deduced amino acid sequences of plastid and mitochondrial Protox in wild type and the herbicide-resistant YZI-1S cells are the same. The level of plastid Protox mRNA was the same in both wild type and YZI-1S cells, whereas the level of mitochondrial Protox mRNA YZI-1S cells was up to 10 times the level of wild type cells. Wild type cells observed by fluorescence microscopy, emitted strong autofluorescence from chlorophyll. Only a weak fluorescence signal was observed from chlorophyll in YZI-1S cells grown in Protox inhibitor S23142. Staining with DiOC6 indicated that there is no visible difference in the number or strength of fluorescence between wild type and YZI-1S mitochondria. Electron micrography of YZI-1S cells showed that in contrast to wild type cell, the chloroplasts of YZI-1S cells grown in the presence of S23142 had no grana stacking. These results suggest that the herbicide resistance of YZI-1S cells is due to the overproduction of mitochondrial Protox.
植物化学調節学会の論文
1998-10-01

D-44 YZI-1S細胞におけるProtoporphyrinogen oxidase阻害型除草剤抵抗性の分子解析

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