SE161:/S1/M2/D1

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Sample Set Information

ID TSE1320
Title Dissection of genotype-phenotype associations in rice grains using metabolome quantitative trait loci analysis.
Description A comprehensive and large-scale metabolome quantitative trait loci (mQTL) analysis was performed to investigate the genetic backgrounds associated with metabolic phenotypes in rice grains. The metabolome dataset consisted of 759 metabolite signals obtained from the grains of 85 lines of rice (Oryza sativa, Sasanishiki x Habataki back-crossed inbred lines). Metabolome analysis was performed using four mass spectrometry pipelines to enhance detection of different classes of metabolites. This mQTL analysis of a wide range of metabolites highlighted an uneven distribution of 802 mQTLs on the rice genome, as well as different modes of metabolic trait (m-trait) control among various types of metabolites. The levels of most metabolites within rice grains were highly sensitive to environmental factors, but only weakly associated with mQTLs. Coordinated control was observed for several groups of metabolites, such as amino acids linked to the mQTL hotspot on chromosome 3. For flavonoids, m-trait variation among the experimental lines was tightly governed by genetic factors that alter the glycosylation of flavones. Many loci affecting levels of metabolites were detected by QTL analysis, and plausible gene candidates were evaluated by in silico analysis. Several mQTLs profoundly influenced metabolite levels, providing insight into the control of rice metabolism. The genomic region and genes potentially responsible for the biosynthesis of apigenin-6,8-di-C-α-l-arabinoside are presented as an example of a critical mQTL identified by the analysis.
Authors Matsuda F, Okazaki Y, Oikawa A, Kusano M, Nakabayashi R, Kikuchi J, Yonemaru J, Ebana K, Yano M, Saito K.
Reference Plant J. 2012 May;70(4):624-36. doi: 10.1111/j.1365-313X.2012.04903.x. Epub 2012 Feb 10.
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Sample Information

ID S1
Title Rice
Organism - Scientific Name Oryza sativa
Organism - ID NCBI taxonomy:4530
Compound - ID
Compound - Source
Preparation The plant population consisted of 85 back‐crossed inbred lines derived from the cross Sasanishiki/Habataki//Sasanishiki///Sasanishiki (Sasanishiki x Habataki) (Nagata et al., 2002b). Seeds from the experimental lines were grown in a paddy field at the National Institute of Agrobiological Sciences (Tsukuba, Japan) in 2005 and 2007, employing similar cultivation schedules. The seeds of the 2005 and 2007 harvests were used for metabolome analysis. One hundred dehulled seeds obtained from whole seeds harvested from 10 independent plants were ground to a fine powder using an MM300 mixer mill (Retsch, http://www.retsch.com/) at 20Hz for 2min in a stainless steel grinding vessel. The powder was divided between small sample tubes (50–100mg) under nitrogen, and the samples were stored at −80°C until analysis.
Sample Preparation Details ID
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Analytical Method Information

ID M2
Title GC‐TOF‐MS
Method Details ID MS2
Sample Amount 1 μL
Comment

Analytical Method Details Information

ID MS2
Title GC‐TOF‐MS
Instrument GC:Agilent 6890N gas chromatograph (Agilent Technologies, Wilmingston, USA) MS:Pegasus III TOF mass spectrometer (LECO, St. Joseph, MI, USA)
Instrument Type
Ionization EI
Ion Mode Positive
Description Sample metabolomic data were acquired using four time-of-flight mass spectrometers (TOF-MS), including CE-TOF-MS for analysis of polar metabolites, GC-TOF-MS for analysis of primary metabolites, LC-Q-TOF-MS for analysis of secondary metabolites and LC-IT-TOF-MS for analysis of lipids. Analysis was performed in triplicate, and the samples were extracted from each pipeline using optimized methods.


The experimental procedures for the GC‐, CE‐ and LC‐IT‐TOF‐MS pipelines were as reported previously (Kusano et al., 2007b; Okazaki et al., 2009; Oikawa et al., 2011).

Comment_of_details Kusano et al., J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Aug;855(1):71-9. Epub 2007 May 16.


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Data Analysis Information

ID D1
Title Data Processing and Analysis (GC-TOF-MS)
Data Analysis Details ID DS2
Recommended decimal places of m/z
Comment


Data Analysis Details Information

ID DS2
Title Data processing and analisys (GC-TOF-MS)
Description The experimental procedures for the GC-, CE- and LC-IT-TOF-MS pipelines were as reported previously (Kusano et al., 2007b; Okazaki et al., 2009; Oikawa et al., 2011).

For GC-TOF-MS analysis, metabolite levels were determined by a single analysis due to technical issues. For GC- and CE-TOF-MS data, a total of 84 and 38 rice seed metabolites, respectively, were identified by comparing the chromatographic behaviour of the metabolites with that of standard compounds.

Comment_of_details Kusano et al., J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Aug;855(1):71-9. Epub 2007 May 16.


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