Metabolite fingerprinting in transgenic Nicotiana tabacum altered by the Escherichia coli glutamate dehydrogenase gene.
Metabolite fingerprinting in transgenic Nicotiana tabacum altered by the Escherichia coli glutamate dehydrogenase gene.
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DOI：
10.1155/jbb.2005.198
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发表时间：
2005-06-30
影响因子：
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通讯作者：
Lightfoot DA
中科院分区：
文献类型：
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作者：
Mungur R;Glass AD;Goodenow DB;Lightfoot DA
由于存在大约20万种植物化学物质，识别那些具有生物医学意义的植物化学物质是一项猛犸的任务。在后基因组时代，将代谢物指纹、丰度和图谱与基因型相关联也是一项艰巨的任务。使用傅立叶变换离子回旋共振质谱（FT-ICR-MS）的离子分析可以提供一种高通量的方法来测量推断的代谢组的基因型依赖性，如果可以建立可重复的技术。离子图谱推断代谢物指纹图谱为副产物。我们使用FT-ICR-MS衍生的离子分析来检查gdhA（谷氨酸脱氢酶（GDH; EC 1.4.1.1））转基因烟草（烟草）进行改变的谷氨酸、氨基酸和碳代谢，这从根本上改变植物生产力。通过标记氨基酸组分和FT-ICR-MS分析代谢产物，分析了gdhA表达、谷氨酸代谢和植物表型之间的因果关系。转gdhA基因植株13 N标记的谷氨酸和谷氨酰胺显著增加。 FT-ICR-MS检测到2 012个离子，在2至4个电离方案中重现。根中有283种离子，叶中有98种离子，由于测量的GDH活性，它们的丰度似乎发生了显着变化。 约58%的离子不能用于推断相应的代谢产物。从42%的离子推断已知的代谢产物，我们发现某些氨基酸，有机酸和糖增加，一些脂肪酸减少。转基因引起增加铵同化和可检测的离子变异。32个具有生物医学意义的化合物被GDH大量改变，包括9个已知的致癌物和14个潜在的药物。因此，GDH转基因可能会为烟草等作物带来新的用途。
With about 200 000 phytochemicals in existence, identifying those of biomedical significance is a mammoth task. In the postgenomic era, relating metabolite fingerprints, abundances, and profiles to genotype is also a large task. Ion analysis using Fourier transformed ion cyclotron resonance mass spectrometry (FT-ICR-MS) may provide a high-throughput approach to measure genotype dependency of the inferred metabolome if reproducible techniques can be established. Ion profile inferred metabolite fingerprints are coproducts. We used FT-ICR-MS-derived ion analysis to examine gdhA (glutamate dehydrogenase (GDH; EC 1.4.1.1)) transgenic Nicotiana tabacum (tobacco) carrying out altered glutamate, amino acid, and carbon metabolisms, that fundamentally alter plant productivity. Cause and effect between gdhA expression, glutamate metabolism, and plant phenotypes was analyzed by labeling of amino acid fractions, and by FT-ICR-MS analysis of metabolites. The gdhA transgenic plants increased 13N labeling of glutamate and glutamine significantly. FT-ICR-MS detected 2 012 ions reproducible in 2 to 4 ionization protocols. There were 283 ions in roots and 98 ions in leaves that appeared to significantly change abundance due to the measured GDH activity. About 58% percent of ions could not be used to infer a corresponding metabolite. From the 42% of ions that inferred known metabolites we found that certain amino acids, organic acids, and sugars increased and some fatty acids decreased. The transgene caused increased ammonium assimilation and detectable ion variation. Thirty-two compounds with biomedical significance were altered in abundance by GDH including 9 known carcinogens and 14 potential drugs. Therefore, the GDH transgene may lead to new uses for crops like tobacco.