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.