SE156:/S1/M5

Sample Set Information

ID	TSE1312
Title	Metabolic Profiling of Developing Pear Fruits Reveals Dynamic Variation in Primary and Secondary Metabolites, Including Plant Hormones.
Description	Metabolites in the fruits of edible plants include sweet sugars, visually appealing pigments, various products with human nutritional value, and biologically active plant hormones. Although quantities of these metabolites vary during fruit development and ripening because of cell division and enlargement, there are few reports describing the actual dynamics of these changes. Therefore, we applied multiple metabolomic techniques to identify the changes in metabolite levels during the development and ripening of pear fruits (Pyrus communis L. ‘La France’). We quantified and classified over 250 metabolites into six groups depending on their specific patterns of variation during development and ripening. Approximately half the total number of metabolites, including histidine and malate, accumulated transiently around the blooming period, during which cells are actively dividing, and then decreased either rapidly or slowly. Furthermore, the amounts of sulfur-containing amino acids also increased in pear fruits around 3–4 months after the blooming period, when fruit cells are enlarging, but virtually disappeared from ripened fruits. Some metabolites, including the plant hormone abscisic acid, accumulated particularly in the receptacle prior to blooming and/or fruit ripening. Our results show several patterns of variation in metabolite levels in developing and ripening pear fruits, and provide fundamental metabolomic data that is useful for understanding pear fruit physiology and enhancing the nutritional traits of new cultivars.
Authors	Oikawa A, Otsuka T, Nakabayashi R, Jikumaru Y, Isuzugawa K, Murayama H, Saito K, Shiratake K.
Reference	PLoS One. 2015 Jul 13;10(7):e0131408. doi: 10.1371/journal.pone.0131408. eCollection 2015.
Comment

PubMed

Sample Information

ID	S1
Title	Pear fruits (Pyrus communis L. ‘La France’)
Organism - Scientific Name	Pyrus communis L.
Organism - ID	NCBI taxonomy:23211
Compound - ID
Compound - Source
Preparation	Pear fruits (Pyrus communis L. ‘La France’) were harvested from a private orchard in Yamagata Prefecture, Japan (lat. 38°11ʹN and long. 140°28ʹE) in 2010 and 2011 with the permission of the owner. Receptacles were removed at 2 weeks before blooming (2WBB), 1 week before blooming (1WBB), the time of blooming (B), and 2 weeks after blooming (2WAB). Furthermore, peeled and deseeded fruits were collected at 1 month after blooming (1MAB), 2 months after blooming (2MAB), 3 months after blooming (3MAB), 4 months after blooming (4MAB), and the time of harvesting (H; 5 months after blooming) and 1 month after harvesting (1MAH; ripened fruits) and used for subsequent metabolomic analyses. These samples were transferred from the orchard to the laboratory as quickly as possible and were then frozen in liquid nitrogen. The frozen samples were weighed and then crushed using a homogenizer. The crushed pear fruits were placed in conical tubes and stored at –80°C until extraction. At least three biological replicates were prepared for each sample.
Sample Preparation Details ID
Comment

Analytical Method Information

ID	M5
Title	Analysis of protein-derived amino acids
Method Details ID	MS5
Sample Amount
Comment

Analytical Method Details Information

ID	MS5
Title	Analysis of protein-derived amino acids
Instrument	CE:Agilent CE capillary electrophoresis system (Agilent Technologies) TOF-MS:Agilent G3250AA LC/MSD TOF system (Agilent Technologies) CE-MS:Agilent G1603A
Instrument Type
Ionization	ESI
Ion Mode	positive and negative
Description	Crushed and frozen samples (500 mg) were resuspended in 1 mL of 50 mM phosphate buffer (pH 7.0), maintained on ice for 30 min, and then centrifuged (12,000 × g, 30 min, 4°C). The supernatant (400 μL) was mixed with 1.2 mL cold acetone and placed in a freezer at –30°C for 120 min. After centrifugation of the mixture (12,000 × g, 5 min, 4°C), the supernatant was carefully discarded and the pellet was suspended in 600 μL of water. Performic acid reagent was prepared by mixing 30% (w/w) H2O2 and 98% (w/w) formic acid at a ratio of 1 to 9. The solution was mixed for 1 h at room temperature. The reagent (12.5 mL) was added to the sample solution (600 μL) and refrigerated at 4°C for 16 h. Most of the reagent was then removed under reduced pressure using a rotary evaporator (N-1000; EYELA, Tokyo, Japan). After evaporation, the sample was resuspended in 15 mL of 6 N HCl and then transferred to a screw-capped tube. The tube was placed in an oil bath (HOA-50A; ASONE, Tokyo, Japan) at 130°C for 20 h. The reagent was again removed under reduced pressure on a rotary evaporator and dissolved in 1 mL of MeOH. A total of 500 μL of this MeOH solution was used for CE-TOF MS analysis as described above. Total protein was quantified by the Bradford method.
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