Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA214029

Local Sample ID:	2_EA_1-pos
Subject ID:	SU002325
Subject Type:	Plant
Subject Species:	Camelina Sativa
Age Or Age Range:	10 days after fertilization
Species Group:	Developing seeds

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Treatment:

Treatment ID:	TR002337
Treatment Summary:	Plants were grown in greenhouses with day/night temperature maintained at 22/20°C, 40-50% relative humidity, and 16h day/8h night photoperiod. Intact siliques during the seed filling growth stage (15 days after fertilization) were excised and placed in sterile media containing a modified Linsmaier and Skoog medium23,24 with Gamborg’s vitamins (Sigma) and 5 mM MES buffer adjusted to pH 5.8. Fifty mM [U-13C6]glucose was used as labeled substrate, and the composition of the remaining carbon and nitrogen sources represented maternal phloem composition to minimize metabolic perturbation and to maintain pseudo in vivo conditions as previously described25. Silique culturing was performed in a 96-well plate with 0.3 mL of medium and a single silique per well, under continuous light (250 µmol m-2 s-1). Tissue was collected and flash frozen immediately after each time point (2, 4, 8, 16 and 32h). Uncultured siliques excised from the maternal plant were used as unlabeled (0h) controls. Frozen tissue was sectioned, on top of dry ice, to excise embryo from the siliques and to separate cotyledons from the embryo axis. Cotyledon samples were extracted and analyzed for lipids in triplicates.

Treatment ID:

TR002337

Treatment Summary:

Plants were grown in greenhouses with day/night temperature maintained at 22/20°C, 40-50% relative humidity, and 16h day/8h night photoperiod. Intact siliques during the seed filling growth stage (15 days after fertilization) were excised and placed in sterile media containing a modified Linsmaier and Skoog medium23,24 with Gamborg’s vitamins (Sigma) and 5 mM MES buffer adjusted to pH 5.8. Fifty mM [U-13C6]glucose was used as labeled substrate, and the composition of the remaining carbon and nitrogen sources represented maternal phloem composition to minimize metabolic perturbation and to maintain pseudo in vivo conditions as previously described25. Silique culturing was performed in a 96-well plate with 0.3 mL of medium and a single silique per well, under continuous light (250 µmol m-2 s-1). Tissue was collected and flash frozen immediately after each time point (2, 4, 8, 16 and 32h). Uncultured siliques excised from the maternal plant were used as unlabeled (0h) controls. Frozen tissue was sectioned, on top of dry ice, to excise embryo from the siliques and to separate cotyledons from the embryo axis. Cotyledon samples were extracted and analyzed for lipids in triplicates.