| Project/Area Number |
18380066
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied biochemistry
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| Research Institution | Kyoto University |
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Principal Investigator |
NISHIOKA Takaaki Kyoto University, GRADUATE SCHOOL OF AGRICULTURE, PROFESSOR. (80026559)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIHARA Atsushi KYOTO UNIVERSITY, GRADUATE SCHOOL OF AGRICULTURE, ASSISTANT PROFESSOR (80281103)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥16,480,000 (Direct Cost: ¥14,800,000、Indirect Cost: ¥1,680,000)
Fiscal Year 2007: ¥7,280,000 (Direct Cost: ¥5,600,000、Indirect Cost: ¥1,680,000)
Fiscal Year 2006: ¥9,200,000 (Direct Cost: ¥9,200,000)
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| Keywords | metabolic flux analysis / metabolic control analysis / Monte Carlo simulations / phenvlpropanoid pathway / elicitor / potato / kinetic network modelling / pulse-feeding experiment / 2次代謝 / 抗菌性物質 / メタボローム解析 / シロイヌナズナ |
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| Research Abstract |
In the present study we have developed a method that allows dynamic flux analysis without a priori kinetic knowledge in the secondary metabolism of plants. This method was developed and validated using the pulse-feeding experimental data, in which incorporation of exogenously applied L-phenylalanine-d5 into seven phenylpropanoid metabolites in potato tubers was determined. After identification of the topology of the metabolic network of these biosynthetic pathways, the system was described by dynamic mass balances in combination with power-law kinetics. As a next step, the exponents of the power-law kinetics were identified and then kept at fixed values during further analysis. The model was tested for statistical reliability using Monte Carlo simulations. Most fluxes could be identified with high accuracy. The two test cases, control and after elicitation, were clearly distinguished, and with elicitation fluxes to N-p-coumaroyloctopamine (pCO) and N-p-coumaroyltyramine (pCT) increased significantly, whereas those for chlorogenic acid (CGA) and p-coumaroylshikimate decreased significantly. According to the model, increases in the first two fluxes were caused by induction/derepression mechanisms. The decreases in the latter two fluxes were caused by decreased concentrations of their substrates, which in turn were caused by increased activity of the pCO- and pCT-producing enzymes. Flux-control analysis showed that, in most cases, flux control was changed after application of elicitor. Thus the results revealed potential targets for improving actions against tissue wounding and pathogen attack.
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