Tung et al_Evolution_2018_DryadData

This .xlsx file has 9 worksheets. The name of each worksheet describes the kind of data that is in it. These are: 1. Activity data_6h_without food 2. Activity data_6h_with food 3. Activity data_24h_with food 4. Exploration data 5. Aggression data 6. Dry weight data 7. Fecundity data 8. Longevity data 9. Metabolites VB refers to the dispersal selected populations, while VBC refers to the corresponding ancestry-matched controls. Thus, VB1 and VBC1 are related to each other by ancestry, were assayed together, and therefore treated as blocks in the analysis

OPLS-DA analysis for S<i>u</i> and I<i>u</i> treatments.

<p>OPLS-DA (a) score plot and (b) loading plot derived from ¹H NMR spectra of S<i>u</i> and I<i>u</i> treatments, with the section above 0 in the loading plot representing metabolites higher in the I<i>u</i> treatment and the section below 0 in the loading plot representing metabolites higher in the S<i>u</i> treatment and (c) S-line plot visualizing differences between I<i>u</i> and S<i>u</i> populations.</p

OPLS-DA analysis for S<i>u</i> and I<i>u</i> treatments.

<p>OPLS-DA (a) score plot and (b) loading plot derived from ¹H NMR spectra of S<i>u</i> and I<i>u</i> treatments, with the section above 0 in the loading plot representing metabolites higher in the I<i>u</i> treatment and the section below 0 in the loading plot representing metabolites higher in the S<i>u</i> treatment and (c) S-line plot visualizing differences between I<i>u</i> and S<i>u</i> populations.</p

Polar dendrogram.

<p>Polar dendrogram showing differences between all the I treatments (I<i>u</i>, I<i>i</i> and I<i>s</i>) and all the S treatments (S<i>u</i>, S<i>i</i> and S<i>s</i>). Each treatment is an average of five replicates with each replicate consisting of 20 flies.</p

Metabolites, their chemical shift values and BH corrected p-values for all pair wise comparisons, with significant metabolites (p < 0.05) for that particular comparison marked in bold.

<p>↑ (up) or ↓ (down) arrows indicate significant increase or decrease in concentration of metabolites respectively, in first treatment as compared to second treatment for each pair-wise comparison.</p

OPLS-DA analysis to see the effect of prick injury.

<p>(a) OPLS-DA score plot for I<i>s</i>-I<i>u</i> comparison, (b) OPLS-DA score plot for S<i>s</i>-S<i>u</i> comparison, (c) S-line plot for I<i>s</i>-I<i>u</i> comparison and (d) S-line plot for S<i>s</i>-S<i>u</i> comparison.</p

Experimental design for the NMR metabolomics experiments.

<p>Four independent blocks, each having two regimes I and S were subjected to three treatments. Each treatment had 5 replicates.</p

OPLS-DA analysis to see the effect of bacterial infection.

<p>(a) OPLS-DA score plot for I<i>i</i>-I<i>u</i> comparison, (b) OPLS-DA score plot for S<i>i</i>-S<i>u</i> comparison, (c) S-line plot for I<i>i</i>-I<i>u</i> comparison and (d) S-line plot for S<i>i</i>-S<i>u</i> comparison.</p

Experimental design for the NMR metabolomics experiments.

<p>Four independent blocks, each having two regimes I and S were subjected to three treatments. Each treatment had 5 replicates.</p

Evolution of the metabolome in response to selection for increased immunity in populations of <i>Drosophila melanogaster</i>

<div><p>We used NMR-based metabolomics to test two hypotheses–(i) there will be evolved differences in the metabolome of selected and control populations even under un-infected conditions and (ii) post infection, the metabolomes of the selected and control populations will respond differently. We selected replicate populations of <i>Drosophila melanogaster</i> for increased survivorship (I) against a gram-negative pathogen. We subjected the selected (I) and their control populations (S) to three different treatments: (1) infected with heat-killed bacteria (<i>i</i>), (2) sham infected (<i>s</i>), and (3) untreated (<i>u</i>). We performed 1D and 2D NMR experiments to identify the metabolic differences. Multivariate analysis of the metabolic profiles of the untreated (I<i>u</i> and S<i>u</i>) flies yielded higher concentrations of lipids, organic acids, sugars, amino acids, NAD and AMP in the I<i>u</i> treatment as compared to the S<i>u</i> treatment, showing that even in the absence of infection, the metabolome of the I and S regimes was different. In the S and I regimes, post infection/injury, concentration of metabolites directly or indirectly associated with energy related pathways (lipids, organic acids, sugars) declined while the concentration of metabolites that are probably associated with immune response (amino acids) increased. However, in most cases, the I regime flies had a higher concentration of such metabolites even under un-infected conditions. The change in the metabolite concentration upon infection/injury was not always comparable between I and S regimes (in case of lactate, alanine, leucine, lysine, threonine) indicating that the I and S regimes had evolved to respond differentially to infection and to injury.</p></div