SE141:/S1/M2
From Metabolonote
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Sample Set Information
ID | TSE1242 |
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Title | Impact of clock-associated Arabidopsis pseudo-response regulators in metabolic coordination. |
Description | In higher plants, the circadian clock controls a wide range of cellular processes such as photosynthesis and stress responses. Understanding metabolic changes in arrhythmic plants and determining output-related function of clock genes would help in elucidating circadian-clock mechanisms underlying plant growth and development. In this work, we investigated physiological relevance of PSEUDO-RESPONSE REGULATORS (PRR 9, 7, and 5) in Arabidopsis thaliana by transcriptomic and metabolomic analyses. Metabolite profiling using gas chromatography-time-of-flight mass spectrometry demonstrated well-differentiated metabolite phenotypes of seven mutants, including two arrhythmic plants with similar morphology, a PRR 9, 7, and 5 triple mutant and a CIRCADIAN CLOCK-ASSOCIATED 1 (CCA1)-overexpressor line. Despite different light and time conditions, the triple mutant exhibited a dramatic increase in intermediates in the tricarboxylic acid cycle. This suggests that proteins PRR 9, 7, and 5 are involved in maintaining mitochondrial homeostasis. Integrated analysis of transcriptomics and metabolomics revealed that PRR 9, 7, and 5 negatively regulate the biosynthetic pathways of chlorophyll, carotenoid and abscisic acid, and alpha-tocopherol, highlighting them as additional outputs of pseudo-response regulators. These findings indicated that mitochondrial functions are coupled with the circadian system in plants. |
Authors | Fukushima A, Kusano M, Nakamichi N, Kobayashi M, Hayashi N, Sakakibara H, Mizuno T, Saito K. |
Reference | Proc Natl Acad Sci U S A. 2009 Apr 28;106(17):7251-6. doi: 10.1073/pnas.0900952106. Epub 2009 Apr 9. Erratum in: Proc Natl Acad Sci U S A. 2009 May 26;106(21):8791. |
Comment |
Sample Information
ID | S1 |
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Title | Plant Materials and Growth Conditions |
Organism - Scientific Name | Arabidopsis thaliana |
Organism - ID | NCBI taxonomy 3702 |
Compound - ID | |
Compound - Source | |
Preparation | A. thaliana accession Columbia (Col-0) was used as a WT plant source. Triple-knockout Arabidopsis mutant prr9-10/prr7-11/prr5-11 has been described in ref. 9. Seedling were grown on Murashige and Skoog (392-00591;Wako) with 0.3% gellan gumand 2% sucrose at pH 5.7 under continuous light (LL) or 12-h light/12-h dark (LD) cycles at 22 °C for 18 days. Whole plants were sampled during days 18–19 to reduce possible developmental effects. |
Sample Preparation Details ID | |
Comment |
Analytical Method Information
ID | M2 |
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Title | Design 2 |
Method Details ID | MS1 |
Sample Amount | 167 μg fresh weight of the derivatized extract |
Comment |
Analytical Method Details Information
ID | MS1 |
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Title | GC-TOF MS |
Instrument | GC:Agilent 6890N MS:LECO Pegasus 4D MS system |
Instrument Type | |
Ionization | EI |
Ion Mode | Positive |
Description | Each sample was extracted, derivatized, and analyzed by using gas chromatography–time-of-flight (GC-TOF/MS) as described in ref. 48. See also SI Methods(http://www.pnas.org/cgi/data/0900952106/DCSupplemental/Supplemental_PDF#nameddest=STXT). All the derivatization steps were performed in the vacuum glove box VSC-100 (Sanplatec, Japan) filled with 99.9995% (G3 grade) of dry nitrogen. The analysis of metabolites by GC-TOF/MS was performed as described previously. |
Comment_of_details |