Table 1. | Metabolite exchange between microbiome members produces compounds that influence Drosophila behavior

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Metabolite exchange between microbiome members produces compounds that influence Drosophila behavior

Table 1.

Affiliation details

Yale University, United States; University of Georgia, United States; University of Connecticut, United States
Table 1.

Summary of volatiles detected using GC-MS. Relative abundance of volatiles in the co-culture (S. cerevisiae and A. malorum grown together) compared to the separate-culture mixture (S. cerevisiae and A. malorum grown separately, and their quantities added in during analysis). GC-MS captured volatiles with XAD-4 beads suspended above the cultures during growth; subsequently, beads were methanol-extracted (n = 6 experiments, Table 1—source data 12). Quantification is based on two experiments in which a linear regression was computed with standards (Table 1—source data 34). Quantification is based on beads suspended above the cultures between 84 and 96 hr of culture growth.

DOI: http://dx.doi.org/10.7554/eLife.18855.018

Table 1—source data 1.Extracted ion chromatograms of five metabolites detected by gas chromatography-mass spectrometry (GC-MS) in Table 1.

Extracted ion chromatograms of the five metabolites detected by gas chromatography- mass spectrometry (GC-MS). (A) Schematic depicting the experimental setup (B-F) Representative extracted ion chromatograms from one replicate (out of three total) of one experiment (out of 3–4 total) of m/z values corresponding to major metabolites identified in the experimental conditions along with appropriate standards. Acetic acid (B), isoamyl alcohol (C), isoamyl acetate (D) isobutanol (E), and ethanol (F) were identified as the five major metabolites in the co-culture (S. cerevisiae and A. malorum). Isoamyl alcohol (C), ethanol (E), and isobutanol (F) were identified as the major metabolites in S. cerevisiae grown alone. Extracted ion chromatograms were constructed using the m/z value in the title of each graph. For acetic acid and isobutanol, the m/z value used corresponds to the molecular weight of the molecule. For ethanol, the m/z used corresponds to the molecular weight minus one (hydrogen). For isoamyl alcohol, the m/z used corresponds to the loss of the hydroxyl group (depicted), which may have picked up hydrogen and been lost as water. For isoamyl acetate, the m/z value corresponds to the molecule shown within the graph. In all cases, figures showing the complete mass spectra between the metabolite and standard are found in Table 1—source data 2. Microorganisms were grown 72–96 hr.

DOI: http://dx.doi.org/10.7554/eLife.18855.019

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Table 1—source data 2.Representative spectra of metabolites in Table 1.

Representative spectra of acetic acid (A-B), isoamyl alcohol (C-E), isoamyl acetate (F-G), ethanol (H-J) and isobutanol (K-L) in standard and experimental samples. Standard concentrations are denoted on individual graphs. All mass spectra are one replicate (out of 3–4 experiments with three replicates per experiment).

DOI: http://dx.doi.org/10.7554/eLife.18855.020

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Table 1—source data 3.Linear regression of metabolites using GC-MS in Table 1.

Estimation of volatile quantity using GC-MS. Separate experiments are graphed in panels (A-E) and (F-J). (A-E) Data points represent the value of a single replicate per concentration for each standard. The abundance of a single m/z value at a specific retention time was chosen for each standard. The values were fitted with a linear regression and the equation was used to estimate the concentration of the five metabolites in the experimental samples from the same experiment. (F-J) Data points represent the mean ± SEM of three replicates for a given concentration for each standard. The abundance of a single m/z value at a specific retention time was chosen for each standard. The values were fitted with a linear regression. The equation was used to estimate the concentration of the five metabolites in the experimental samples from the same experiment. When applicable an equation was calculated when the line was forced to go through X,Y = 0,0; these equations were used to calculate the concentrations of isoamyl alcohol, isoamyl acetate, and isobutanol.

DOI: http://dx.doi.org/10.7554/eLife.18855.021

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Table 1—source data 4.Raw spectral abundance data as a function of concentration used for linear regressions in Table 1—source data 3.

DOI: http://dx.doi.org/10.7554/eLife.18855.022

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Identity

Standard confirmation

Relative quantification (co-culture: separate-culture mixture)

Ethanol

Y

5.0–12.6-fold reduced

Isobutanol

Y

7.3–24.7-fold reduced

Isoamyl acetate

Y

unique to co-culture

Isoamyl alcohol

Y

3.6–6.4-fold reduced

Acetic acid

Y

unique to co-culture