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拟南芥ispD和ispE白化突变体中4-磷酸甲基赤藓糖醇途径酶的叶绿体定位及线粒体基因调控

Chloroplast localization of methylerythritol 4-phosphate pathway enzymes and regulation of mitochondrial genes in ispD and ispE albino mutants in Arabidopsis.

作者信息

Hsieh Ming-Hsiun, Chang Chiung-Yun, Hsu Shih-Jui, Chen Ju-Jiun

机构信息

Institute of Plant and Microbial Biology, Academia Sinica, Taipei 11529, Taiwan.

出版信息

Plant Mol Biol. 2008 Apr;66(6):663-73. doi: 10.1007/s11103-008-9297-5. Epub 2008 Jan 31.

Abstract

Plant isoprenoids are derived from two independent pathways, the cytosolic mevalonate pathway and the plastid methylerythritol 4-phosphate (MEP) pathway. We used green fluorescent fusion protein assays to demonstrate that the Arabidopsis MEP pathway enzymes are localized to the chloroplast. We have also characterized three Arabidopsis albino mutants, ispD-1, ispD-2 and ispE-1, which have T-DNA insertions in the IspD and IspE genes of the MEP pathway. Levels of photosynthetic pigments are almost undetectable in these albino mutants. Instead of thylakoids, the ispD and ispE mutant chloroplasts are filled with large vesicles. Impairments in chloroplast development and functions may signal changes in the expression of nuclear, chloroplast and mitochondrial genes. We used northern blot analysis to examine the expression of photosynthetic and respiratory genes in the ispD and ispE albino mutants. Steady-state mRNA levels of nucleus- and chloroplast-encoded photosynthetic genes are significantly decreased in the albino mutants. In contrast, transcript levels of nuclear and mitochondrial genes encoding subunits of the mitochondrial electron transport chain are increased or not affected in these mutants. Genomic Southern blot analysis revealed that the DNA amounts of mitochondrial genes are not enhanced in the ispD and ispE albino mutants. These results support the notion that the functional state of chloroplasts may affect the expression of nuclear and mitochondrial genes. The up-regulation of mitochondrial genes in the albino mutants is not caused by changes of mitochondrial DNA copy number in Arabidopsis.

摘要

植物类异戊二烯来源于两条独立的途径,即胞质甲羟戊酸途径和质体甲基赤藓糖醇4-磷酸(MEP)途径。我们使用绿色荧光融合蛋白分析来证明拟南芥MEP途径的酶定位于叶绿体。我们还鉴定了三个拟南芥白化突变体ispD-1、ispD-2和ispE-1,它们在MEP途径的IspD和IspE基因中存在T-DNA插入。在这些白化突变体中,光合色素水平几乎检测不到。ispD和ispE突变体的叶绿体中没有类囊体,而是充满了大的囊泡。叶绿体发育和功能的受损可能标志着核基因、叶绿体基因和线粒体基因表达的变化。我们使用Northern印迹分析来检测ispD和ispE白化突变体中光合基因和呼吸基因的表达。白化突变体中核编码和叶绿体编码的光合基因的稳态mRNA水平显著降低。相反,编码线粒体电子传递链亚基的核基因和线粒体基因的转录水平在这些突变体中增加或不受影响。基因组Southern印迹分析表明,ispD和ispE白化突变体中线粒体基因的DNA量没有增加。这些结果支持叶绿体的功能状态可能影响核基因和线粒体基因表达的观点。白化突变体中线粒体基因的上调不是由拟南芥线粒体DNA拷贝数的变化引起的。

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