SE147:/S1/M1/D1

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Sample Set Information

ID	TSE1308
Title	Landscape of the lipidome and transcriptome under heat stress in Arabidopsis thaliana.
Description	Environmental stress causes membrane damage in plants. Lipid studies are required to understand the adaptation of plants to climate change. Here, LC-MS-based lipidomic and microarray transcriptome analyses were carried out to elucidate the effect of short-term heat stress on the Arabidopsis thaliana leaf membrane. Vegetative plants were subjected to high temperatures for one day, and then grown under normal conditions. Sixty-six detected glycerolipid species were classified according to patterns of compositional change by Spearman’s correlation coefficient. Triacylglycerols, 36:4- and 36:5-monogalactosyldiacylglycerol, 34:2- and 36:2-digalactosyldiacylglycerol, 34:1-, 36:1- and 36:6-phosphatidylcholine, and 34:1-phosphatidylethanolamine increased by the stress and immediately decreased during recovery. The relative amount of one triacylglycerol species (54:9) containing α-linolenic acid (18:3) increased under heat stress. These results suggest that heat stress in Arabidopsis leaves induces an increase in triacylglycerol levels, which functions as an intermediate of lipid turnover, and results in a decrease in membrane polyunsaturated fatty acids. Microarray data revealed candidate genes responsible for the observed metabolic changes.
Authors	Higashi Y, Okazaki Y, Myouga F, Shinozaki K, Saito K.
Reference	Sci Rep. 2015 May 27;5:10533. doi: 10.1038/srep10533.
Comment

ID	S1
Title	Arabidopsis thaliana
Organism - Scientific Name	Arabidopsis thaliana
Organism - ID	NCBI taxonomy:3702
Compound - ID
Compound - Source
Preparation	Arabidopsis thaliana ecotype Columbia (Col-0) and Nossen were used. Arabidopsis seeds were surface-sterilised and sown on an agar-solidified Murashige and Skoog medium containing 0.5% (w/v) sucrose. Plants were grown at 22 °C under a 16-h-light/8-h-dark cycle. Fourteen-day-old Col-0 and Nossen plants at about 3 h after the onset of the light phase were subjected to heat stress at 22 °C (control), 30 °C, 34 °C or 38 °C for one day under a continuous light condition (biological replicate N = 4) (Supplementary Fig. S1). For recovery experiments, 18-day-old Col-0 and 14-day-old Nossen plants were subjected to heat stress at 38 °C for one day under the 16/8 h light-dark cycle, and then returned to 22 °C and grown for one day and 2 days longer (biological replicate N = 3, 4, 8, or 12). Aerial parts were harvested at about 3 h after the onset of light phase.
Sample Preparation Details ID
Comment

ID	MS1
Title	LCMS-IT-TOF
Instrument	LC, Shimadzu LC-20AD system; MS, Shimadzu LCMS-IT-TOF
Instrument Type
Ionization	ESI
Ion Mode	negative
Description	Crude lipid was extracted from the aerial parts according to the method of Okazaki et al.. The dried chloroform extract was dissolved in 160 μL ethanol for LC-MS analysis. The conditions for LC-MS analysis were described in the same paper. Lipid data were obtained from 77 distinct plant samples that included 6 environmental conditions and 2 ecotypes, in which the plant samples were prepared from 6 individual growth periods.
Comment_of_details

ID	D1
Title	Data Processing and Analysis
Data Analysis Details ID	DS1
Recommended decimal places of m/z
Comment

ID	DS1
Title	Data processing and analisys
Description	Levels of individual lipid species in every sample preparation were normalised to the sum of peak areas of lipid species within the same lipid class in samples grown at 38 °C for one day (1d38C). The scaled levels of the 66 lipid species, which were detected under the all conditions and ecotypes, were calculated to show overview as a heat map, where levels of individual lipid species were subtracted by the mean of values of the lipid species among the 77 samples, then divided by the standard deviations for values of the lipid species among the 77 samples as described in Vu et al.27. Spearman’s correlation coefficients were calculated among 12 variables, which were expressed as the average of each condition (22 °C control (1d22C and 1d22C+2d22C), 30 °C for one day (1d30C), 34 °C for one day (1d34C), 1d38C, 38 °C for one day then 22 °C for one day (1d38C+1d22C), and 38 °C for one day then 22 °C for 2 days (1d38C+2d22C)) for each ecotype (Col-0 (C) and Nossen (N)). Hierarchical clustering of the lipid species was conducted by the complete linkage method based on Euclidean distance (k = 10) by the software R. A heat map was constructed to depict the log2 of the ratio of the average of each stress condition to that of the 22 °C control. Significant differences in individual values were investigated by Welch’s t test (two-sided).
Comment_of_details