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Diurnal Dehydration Effects on Germination & Metabolism in Arabidopsis Ecotypes

This study examines the stratified seed germination of four Arabidopsis ecotypes (Cvi, Ler, C24, Col-0) subjected to three diurnal dehydration cycles. Supplementary figures illustrate significant alterations in fatty acid profiles, malondialdehyde (MDA) content, and protein levels following hydration-dehydration cycles. Additionally, Partial Least Squares Discriminant Analysis (PLS-DA) highlights notable ecotypic differences in metabolic responses over time. Key metabolites are analyzed, advancing our understanding of germination dynamics influenced by dehydration stress.

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Diurnal Dehydration Effects on Germination & Metabolism in Arabidopsis Ecotypes

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  1. Supplementary Figure S1. Visualization of the stratified-seed germination after three diurnal cycles of dehydration (Dh 1, Dh 2, Dh 3) compared with seeds germinated following stratification (St) for four Arabidopsis ecotypes

  2. Cvi Ler C24 Col-0 16:0 Percentage of total FAs (%) 18:0 18:1n-9 18:2n-6 18:3n-3 20:1n-9 Supplementary Figure S2. Fatty acid profile, as percentage of total FAs of lipids extracted from seeds of four Arabidopsis ecotypes through three continuous hydration-dehydration cycles, Error bars represent mean ± SE.

  3. MDA content (μmol/g) Cvi Ler C24 Col-0 Supplementary Figure S3. Malondialdehyde (MDA) equivalents (µmol/g) recovered from Arabidopsis seeds of four ecotypes following repetitive dehydrations. Error bars represent mean ± SE of 3 independent measurements.

  4. Supplementary Figure S4. Partial Least Squares - Discriminant Analysis (PLS-DA) of ecotypic differences across Hy-Dh cycles. 3D score plot between the selected PCs and corresponding explained variances are shown.

  5. Hy 4 Hy 3 Dh 3 Hy 2 Dh 1 Dh 2 Dry Hy 1 St Ler Ler Ler Ler Ler Col-0 Col-0 Col-0 Col-0 Col-0 Ler Col-0 Cvi Cvi Cvi Cvi Cvi Cvi Ler C24 C24 C24 C24 Ler Ler Col-0 C24 Col-0 Col-0 C24 Cvi Cvi Cvi C24 C24 C24 0 0.4 0.8

  6. Supplementary Figure S5. False scale color visualization of significant (p<0.05, ANOVA) metabolic difference among ecotypes at each experimental time point tested. False scale color represents the relative intensity normalized by the median of the selected metabolites across all the time points followed by log transformation (base 10), and expressed as the mean of three independent replicas. Abbreviation : Glc-1,6-AH, glucose-1,6-anhydro,beta; Myo-inositol-1-P, myo-inositol-1-phosphate; PME, phosphorate-monomethyl ester; Glycerol-3-P, glycerol-3-phosphate; GPG, glycerophosphoglycerol; Gluconate-1,5-L, gluconate-1,5-lactone.

  7. C24 Col-0 Cvi Ler Cvi Ler Col-0 C24 Supplementary Figure S6. Relative yield of the reduced Triphenyl tetrazolium chloride (TTC) by TCA/acetone extraction. Absorbance was plate assayed at 485 nm. The extraction absorbance was measured (left). TTC reduction is reflected by the change in color of the extract (right). Values are presented by the mean of five independent preparations ± SE.

  8. Ler Cvi C24 Col-0 Supplementary Figure S7. Protein content of seeds after three hydration-dehydration cycles compared with that after stratification. Values are means of four independent preparations. Error bars represent mean ± SE

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