SE198:/S12/M91/D9

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Sample Set Information

ID SE198
Title Untargeted metabolome analysis of 15N- or 34S-labeled plants (lettuce) / 15Nまたは34S全標識した植物のLC-MSメタボローム解析(レタス)
Description Untargeted metabolome analyses of lettuce were performed using liquid chromatography – mass spectrometry (LC-MS). This data acquisition aims at an improvement of peak annotation by using plant samples that were fully labeled with stable isotope 15N or 34S.

Two lettuce varieties, Leaf Lettuce Green and King Crown were used.

The information of the peaks detected in Leaf Lettuce Green was available at the Plant Metabolome Repository website (http://metabolites.in/plants).


15Nおよび34Sで全標識した植物試料を用いることで、液体クロマトグラフィー-質量分析(LC)で検出される化合物ピークの推定を向上させることを目的として、ノンターゲットメタボローム解析を行った。

植物試料として、二種類のレタス(リーフレタスグリーンおよびキングクラウン)を用いた。

リーフレタスグリーンで検出されたピークは、植物メタボロームレポジトリ (http://metabolites.in/plants) から公開されている。

Authors Sakurai N1,2, Suda K1, Akimoto N1, Hoshi K1, Osawa S1, Ikeda C1, Ozawa K1, Yamada M1, Muneto R1, Shibata D1 (1 Kazusa DNA Research Institute, 2 National Institute of Genetics), Contact: sakurai AT nig.ac.jp (replace AT with @)


櫻井望1,2、須田邦裕1、秋元奈弓1、星久美1、大澤祥子1、池田千晶1、小澤馨史1、山田学1、宗藤玲子1、柴田大輔1(1 かずさDNA研究所, 2 国立遺伝学研究所)

Reference
Comment

Sample Information

ID S12
Title Grand Rapids (Leaf Lettuce Green) (Control), freeze dried
Organism - Scientific Name Lactuca sativa L. var crispa
Organism - ID NCBI taxonomy: 466611
Compound - ID
Compound - Source
Preparation
Sample Preparation Details ID SS1
Comment

Sample Preparation Details Information

ID SS1
Title Preparation of the samples
Description Seeds of lettuce “Grand Rapids (Leaf Lettuce Green)” (Lactuca sativa L. var crispa) and “King Crown” (Lactuca sativa L. var capitata) were purchased from SAKATA SEED CORPORATION (Kanagawa, Japan). The seeds were germinated on vermiculite as a support medium in pots. The plants were grown in a growth chamber (12 h light/12 hr dark, 22 C) for 33 days and then on a cultivation shelf (16 hr light/ 8 hr dark, 20-28 C). The leaves were harvested 77 and 83 days after germination for Leaf Lettuce Green and King Crown, respectively. The plants were fertilized using a solution containing 3 mM KNO3, 1 mM Ca(NO3)2, 0.5 mM MgSO4, 0.5 mM NH4H2PO4, 0.05 mM Fe(III)-ethylenediamine-N,N,N’,N’-tetraacetic acid (EDTA), 49 uM H3BO3, 9 uM MnSO4, 0.8 uM ZnSO4, 0.3 uM CuSO4, and 0.1 uM Na2MoO4. For the stable isotope labeling, KNO3, Ca(NO3)2, and NH4H2PO4 were replaced with their 15N labeled chemicals (99.2-99.8 Atom%) and MgSO4 was replaced with its 34S labeled chemical (98 Atom%). labeled chemicals were purchased from Shoko Science Co., Ltd. (Yokohama, Japan). The plants were fertilized properly using the same amount of the solution in one time for control, 15N-labeled, and 34S-labeled plants. In total, 2,530 and 3,040 mL of the solution was used for growing three individuals of Leaf Lettuce Green and King Crown, respectively. The shoot of three individuals for each treatment was harvested, mixed, weighed, and homogenated using mortar and pestle under liquid nitrogen into a fine powder. The sample was stored at -80 C until use. A part of the powdered sample was freeze-dried and stored at room temperature.
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Analytical Method Information

ID M91
Title PSEUDO: Negative, A set of analyses for valid peak selection
Method Details ID
Sample Amount
Comment This virtual item represents a set of several analyses used for the peak alignment and characterization.

Data Analysis Information

ID D9
Title Valid peak selection by alignment and characterization
Data Analysis Details ID DS2
Recommended decimal places of m/z default
Comment


Data Analysis Details Information

ID DS2
Title Selection of valid peaks by alignment using PowerGetBatch
Description The detected peaks in the samples with high- and low-resolution analyses and in the mock samples were aligned using PowerGetBatch software for each positive and negative mode. The reproducibly detected peaks in the samples and absent in the mock samples were selected as valid peaks. The selection was performed manually using Microsoft Excel with consideration of the analytical replications of the sample. The most intense and major patterns of the MS2 and MS3 spectra were selected among the alignment results for each peak. Searching candidate compounds in compound databases and prediction of flavonoid aglycones using FlavonoidSearch system for valid peaks were performed as described in the annotation details AM1.


The samples used for the alignment are shown in the Plant Metabolome Repository website (http://metabolites.in/plants/dl/19812).


The alignment parameter is available at the Plant Metabolome Repository website (http://metabolites.in/plants/data/PGB_params.zip).

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