Table 2. | 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 2.

Affiliation details

Yale University, United States; University of Georgia, United States; University of Connecticut, United States
Table 2.

Estimated concentrations of key metabolites in the co-culture using SPME GC-MS. Estimated concentrations of differentially concentrated or unique metabolites in the co-culture. Linear regression equations (Lin. reg. eqs. 1 and 2) were estimated from individual experiments in which peak areas of different concentrations of metabolites were fitted with a linear regression (Table 2—source data 2, 3, 5 and 6). Normalized peak areas correspond to the specified metabolites in co-cultures containing S. cerevisiae and A. malorum. Separate estimates were derived from a normalized peak area estimated from a single experiment (co-culture and standard samples were from a run with similar internal standard signal) or from the mean normalized peak area estimated from all experiments (co-cultures were run over four days, standards were run on two days). The final estimated concentration was an average of all estimated concentrations (n = 4 estimates (two from each standard regression equation times two estimates of the normalized peak area), except for methyl acetate, n = 2 estimates). The estimated concentrations (except acetoin) were added to the co-culture containing A. pomorum adhA (Figure 4C). *Ethyl acetate, acetic acid, and acetoin concentrations were estimated from standards (Table 2—source data 1 and 4).

DOI: http://dx.doi.org/10.7554/eLife.18855.029

Table 2—source data 1.Extracted ion chromatograms of differentially emitted or unique metabolites in the co-culture in Table 2.

Extracted ion chromatograms of differentially emitted or unique metabolites in the co-culture according to solid phase microextraction gas chromatography-mass spectrometry (SPME GC-MS). Specific metabolites are displayed above each panel. For each panel, the left-most plot compares the co-culture containing S. cerevisiae and A. malorum to S. cerevisiae grown alone, A. malorum grown alone, or media (AJM [apple juice medium]); the right-most plot compares the co-culture containing S. cerevisiae and A. pomorum wild-type to the co-culture containing S. cerevisiae and A. pomorum adhA, since A. pomorum adhA is required for Drosophila co-culture preference (Figure 5A). The two plots within the same panel contain the same standard. The y-axis for each plot is the ion current for a m/z value that discriminates the metabolite of interest over a specific retention time window. The following m/z values were chosen for each metabolite based on standards or, in the cases of putative and unknown metabolites (I and J) were chosen from the experimental groups: (A) m/z 74.04 (B) m/z 88.08 (C) m/z 73.03 (D) 87.05 (E) 74.02 (F) 104.04 (G) 60.05 (H) 88.05 (I) 101.06 (J) 101.06. Each panel is one representative replicate of 1 experiment (out of 3–5 total replicates in three experiments).

DOI: http://dx.doi.org/10.7554/eLife.18855.030

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Table 2—source data 2.Linear regression of metabolites in defined metabolite mixtures in Table 2.

Normalized peak areas corresponding to metabolites in a defined metabolite mixture (from SPME GC-MS). A linear regression was calculated to quantify the metabolites in the co-culture. Each concentration is from one replicate. A-E and F-I are two separate experiments. Linear regression was used to estimate the concentration of the metabolites in the co-culture containing S. cerevisiae and A. malorum (Table 2) and to complement the co-culture containing A. pomorum adhA (Figure 4C).

DOI: http://dx.doi.org/10.7554/eLife.18855.031

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Table 2—source data 3.Peak area as a function of concentration used to estimate metabolite concentrations in co-cultures in Table 2.

DOI: http://dx.doi.org/10.7554/eLife.18855.032

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Table 2—source data 4.Extracted ion chromatograms of various m/z values used in.

DOI: http://dx.doi.org/10.7554/eLife.18855.033

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Table 2—source data 5.Peak areas as a function of metabolite concentration used in linear regression in Table 2—source data 2A–E.

DOI: http://dx.doi.org/10.7554/eLife.18855.034

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Table 2—source data 6.Peak areas as a function of metabolite concentration used in Table 2—source data 2F–I.

DOI: http://dx.doi.org/10.7554/eLife.18855.035

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Metabolite

Lin. Reg. eq. 1

Lin. Reg. eq. 2

Normalized peak area (single experiment)

Normalized peak area (Average, All experiments)

Estimated concentration (%)

Isobutyl acetate

Y = 4151X − 0.1319

Y = 3252X − 0.07251

0.29

1.16

0.00023

Isoamyl acetate

y = 8158X

Y = 7800X

0.78

3.8

0.00026

2-Phenethyl acetate

Y = 5129X −0.04011

Y = 6972X −0.2013

1.2

1.9

0.00028

2-Methylbutyl acetate acetate

Y = 8995X − 0.05042

Y = 8087X−0.1307

0.56

3.1

0.00023

Methyl acetate

Y = 75.22X+0.004457

NA

0.018

0.040

0.00033

Ethyl acetate

NA

NA

NA

NA

~0.02*

Acetic acid

NA

NA

NA

NA

~3.0*

Acetoin

NA

NA

NA

NA

~0.01*