SUPPLEMENTARY MATERIAL
Supplementary Table S1. Whole dataset produced from untargeted metabolomics carried out in Arabidopsis roots exposed to heat, drought and the combined stress. Compounds are presented with individual intensities and with composite mass spectra (monoisotopic accurate mass/abundance combinations).
Supplementary Table S2. Discriminant metabolites (VIP markers) identified by the OPLS-DA analysis in Arabidopsis roots exposed to heat, drought and combined stress
Supplementary Table S3. Differential metabolites derived from ANOVA and fold-change (FC) analysis (p-value < 0.05, Benjamini multiple testing correction; fold-change threshold FC ≥ 2) in roots metabolomic profiles after stress exposure. These compounds were uploaded into the Omic Viewer Pathway Tool of PlantCyc (Plant Metabolic Network, http://www.plantcyc.org/).
Supplementary Figure S1. Unsupervised hierarchical cluster analysis (Euclidean distance; linkage rule: Ward) carried out from root chemical profiles exposed to heat, drought and the combined stress. Metabolites were obtained by UHPLC-ESI/QTOF-MS untargeted analysis, and their intensities used to build up the fold-change heatmap here provided.
Supplementary Figure S2: Cladogram of LEfSe Biomarker analysis in root and soil. Taxonomic nodes that explain the most variance and are indicative for a certain treatment are coloured by treatment. Each ring represents a taxonomic level starting from phylum (p__) over class (c__), order (o__), family (f__) to genus (g__). The described analysis was performed with LDA score cut-off of 3, but only marker with an LDA score > 4 are depicted in this cladogram to avoid an overcrowded image.
Supplementary Figure S3 : Venn diagram showing the number of features shared between LefSe (red) and ANCOMBC (grey) analyses. Filtered soil and root datasets were aggregated at genus level and analysed by discriminant analysis (LEfSe, grey) with LDA cut-off set to 2. For differential analysis p-values were adjusted for multiple testing using holm-Bonferroni method and p-value cut-off = 0.01 was chosen the define significance.
Supplementary Figure S4 : Plotted effect Size of Biomarkers identified by LefSE analysis for A) root and B) soil with an LDA score over 3. Color indicates the treatment they discriminant for size of dot represent p-value. The marker taxonomic lineage is displayed on 3 levels.
Supplementary Figure S5 : multiblock sPLS-DA (DIABLO). Samples from data blocks metabarcoding and metabolomics are plotted into space spanned by the second and third components. Length of the arrows indicate the distance of each sample from the centroids of both datasets. Short distances show a high level of agreement between blocks. For all samples and treatments the agreement between metabarcoding and metabolomics is high.