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Articles by Yuuri Tsuboi in JoVE
JoVE General
Nükleer Manyetik Rezonans Spektroskopisi ile Çevre metabolomik için düşük yoğunluklu Planktonik topluluklardan Metabolitlerinin Konsantrasyon
1Biosphere Oriented Biology Research Unit, RIKEN Advanced Science Institute, 2Graduate School of Nanobioscience, Yokohama City University, 3Advanced NMR Metabomics Research Team, RIKEN Plant Science Center, 4Graduate School of Bioagricultural Science, Nagoya University
Mikrobiyal planktonik toplulukların metaboliti çıkarımı için bir yöntem sunulmaktadır. Tüm topluluk örnekleme özel hazırlanmış filtreler üzerine filtrasyon sağlanır. Liyofilizasyon sonra, sulu çözünür metabolitleri ekstre edilir. Bu yaklaşım doğal ya da deneysel mikrobiyal toplulukların trans-omik araştırmalar çevre metabolomik uygulaması için izin verir.
Other articles by Yuuri Tsuboi on PubMed
The Ectopic Expression of Phenylalanine Ammonia Lyase with Ectopic Accumulation of Polysaccharide-linked Hydroxycinnamoyl Esters in Internode Parenchyma of Rice Mutant Fukei 71
Both polysaccharide-linked hydroxycinnamoyl esters (PHEs) and lignin are biosynthesized via the phenylpropanoid pathway. In the abnormal internode parenchyma of the rice (Oryza sativa L.) mutant Fukei 71, which has a defective recessive gene (d50), the biosynthesis of lignin and PHEs differs. . The polysaccharide-linked ferulate and p-coumarate have been shown to accumulate to high levels in the irregularly shaped and collapsed internode parenchyma cells of Fukei 71 without an accompanying overaccumulation of lignin as a result of the defective d50 gene. In the present study we demonstrated that in this abnormal parenchyma tissue of Fukei 71 the expression of phenylalanine ammonia lyase (PAL) and glutamine synthetase (GS) were ectopically induced with the ectopic accumulation of PHEs, suggesting that the d50 gene may play a role as a controlling element in the biosynthesis of PHEs during cell-wall formation in the grasses.
3-Deoxy-D-arabino-heptulosonate 7-phosphate Synthase is Regulated for the Accumulation of Polysaccharide-linked Hydroxycinnamoyl Esters in Rice (Oryza Sativa L.) Internode Cell Walls
Polysaccharide-linked hydroxycinnamoyl esters (PHEs) over-accumulate in the internodes of a rice (Oryza sativa L.) mutant, Fukei 71 (F71). This accumulation is accompanied by over-expression of phenylalanine ammonialyase (PAL). In this study, we show that only one member of the 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase (DAHPS) family expresses in close correlation with PAL. Furthermore, substrate availability to DAHPS is promoted by down-regulating the expression of plastidic pyruvate kinase (PKp), a competitor of DAHPS. Since the over-production of PHEs is caused by D50 gene disruption, these results suggest that specific enzymes in the phenylpropanoid and shikimate pathways are coordinately up-regulated. In addition, the results indicate that carbon-flow into the shikimate pathway is modified for the synthesis of PHEs, and is probably controlled by D50.
Top-down Phenomics of Arabidopsis Thaliana: Metabolic Profiling by One- and Two-dimensional Nuclear Magnetic Resonance Spectroscopy and Transcriptome Analysis of Albino Mutants
Elucidating the function of each gene in a genome is important for understanding the whole organism. We previously constructed 4000 disruptant mutants of Arabidopsis by insertion of Ds transposons. Here, we describe a top-down phenomics approach based on metabolic profiling that uses one-dimensional 1H and two-dimensional 1H,13C NMR analyses and transcriptome analysis of albino mutant lines of Arabidopsis. One-dimensional 1H NMR metabolic fingerprinting revealed global metabolic changes in the albino mutants, notably a decrease in aromatic metabolites and changes in aliphatic metabolites. NMR measurements of plants fed with 13C6-glucose showed that the albino lines had dramatically different 13C-labeling patterns and increased levels of several amino acids, especially Asn and Gln. Microarray analysis of one of the albino lines revealed a unique expression profile and showed that changes in the expression of genes encoding metabolic enzymes did not correspond with changes in the levels of metabolites. Collectively, these results suggest that albino mutants lose the normal carbon/nitrogen balance, presumably mainly through lack of photosynthesis. Our study offers an idea of how much the metabolite network is affected by chloroplast function in plants and shows the effectiveness of NMR-based metabolic analysis for metabolite profiling. On the basis of these findings, we propose that future investigations of plant systems biology combine transcriptomic, metabolomic, and phenomic analyses of gene disruptant lines.
Statistical Indices for Simultaneous Large-scale Metabolite Detections for a Single NMR Spectrum
NMR-based metabolomics has become a practical and analytical methodology for discovering novel genes, biomarkers, metabolic phenotypes, and dynamic cell behaviors in organisms. Recent developments in NMR-based metabolomics, however, have not concentrated on improvements of comprehensiveness in terms of simultaneous large-scale metabolite detections. To resolve this, we have devised and implemented a statistical index, the SpinAssign p-value, in NMR-based metabolomics for large-scale metabolite annotation and publicized this information. It enables simultaneous annotation of more than 200 candidate metabolites from the single (13)C-HSQC (heteronuclear single quantum coherence) NMR spectrum of a single sample of cell extract.
