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Figures
- Fig. 1.
Systems of genes/enzymes underlying ethanol metabolism and lipid-derived signaling pathways, along with the regulatory effects of the sterol regulatory element binding protein (dSREBP). The two parallel curved lines represent the plasma membrane. AcCoAS, acetyl-CoA synthetase; ADH, alcohol dehydrogenase; ALDH, acetaldehyde dehydrogenase; DAG, diacylglycerol; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PEth, phosphatidylethanol; PLD, phospholipase D; PPAP, phosphatidate phosphatase.
- Table 1.
Population and temperature effects on ethanol and acetic acid tolerances, enzyme activities and gene expression
Fixed effect Population TRear TExpose (TRear) Pop×TRear Pop×TExpose (TRear) Trait F P F P F P F P F P Acetic acid tolerance 0.07 23.16 −−−− 2.19 0.40 0.44 Ethanol tolerance 12.35 +++ 45.26 ++++ 41.96 **** 0.37 0.77 AcCoAS activity (EC 6.2.1.1) 3.71 28.99 ++++ 3.38 * 1.23 1.86 AcCoAS expression (CG9390) 5.73 + 9.52 ++ 3.25 * 3.26 0.71 CG6432 (putative AcCoAS) 21.59 +++ 10.53 ++ 55.62 **** 2.60 0.14 CG8732 (putative AcCoAS) 2.72 37.76 ++++ 1.43 0.31 0.89 ADH activity (EC 1.1.1.1) 51.81 ++++ 291.2 ++++ 9.88 **** 7.77 ** 1.60 Adh expression (CG3481) 11.58 ++ 0.17 5.38 ** 0.33 1.67 ALDH activity (EC 1.2.1.3) 0.56 3.07 6.27 ** 17.63 **** 0.32 Aldh expression (CG3752) 0.50 90.14 −−−− 1.09 0.63 0.48 Cdpet expression (CG6016) 0.03 24.12 ++++ 4.85 ** 5.07 * 0.60 desatl expression (CG5887) 0.57 18.17 ++++ 1.09 1.66 1.16 Pect expression (CG5547) 1.82 8.24 ++ 0.24 0.60 0.62 PLD activity (EC 3.1.4.4) 0.05 48.02 ++++ 4.76 ** 5.09 * 0.91 Pld expression (CG12110) 1.89 4.62 + 6.95 ** 2.40 0.6 Sply expression (CG8946) 5.65 + 4.82 + 18.98 **** 3.25 0.24 Srebp expression (CG8522) 1.92 6.50 − 1.44 0.94 0.57 wunen expression (CG8804) 0.00 5.99 − 7.63 *** 1.68 0.02 -
The number of symbols indicates the level of significance from mixed model ANOVAs; +, increased values in Tasmania or 26°C reared flies, −, increased values in Innisfail or 15°C reared flies;* significant at *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.
CG numbers are gene identifiers (FlyBase Consortium, 2003).
-
- Fig. 2.
Correlation between ethanol and acetic acid tolerances across species of Drosophila and populations of D. melanogaster. Points represent the lethal dose of toxin causing a 50% male mortality (LD50) in each genetic line. Error bars represent 95% confidence intervals for LD50 values. Australian population data are from flies acclimated to and tested at 15°C. All others are from a survey of tolerances in flies reared and assayed at 24°C.
- Fig. 3.
ADH activities and ethanol tolerances for each Adh genotypic class. Lines are categorized as having only the Adh-F allele (F, N=5 lines), only the Adh-S allele (S, N=5 lines) or having both alleles (F&S, N=10 lines). Data are least square means ± 1 standard error. The inset contains Adh expression data (1/Ct) for the three genotype classes.
- Fig. 4.
Mortality curves for extreme high and low ethanol tolerant lines acclimated to and assayed at 26°C. Symbols represent the two replicate observations for each line at each of five ethanol concentrations. Lines with 95% confidence intervals (horizontal bars) are fits from probit regressions from which we obtained estimates of LD50.
- Fig. 5.
Population and temperature acclimation effects on ethanol tolerance (A), the ethanol catabolic enzymes, ADH (B), AcCoAS (C) and two putative AcCoAS-encoding loci (D). Shown are least square means ± 1 standard error and significant fixed effects from mixed-model ANOVAs. Fold expression changes were calculated from serial dilution standard curves. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.
- Fig. 6.
Phosphatidylethanolamine (PE) biosynthesis pathway (A) and the population and temperature acclimation effects on expression of genes encoding enzymes in this pathway, as well as a fatty acid desaturase (B). Shown are least square means ± 1 standard error and significant fixed effects from mixed-model ANOVAs. Units of expression are (1/Ct)×1000. Fold expression changes were calculated from serial dilution standard curves. CDPET, CDP-ethanolamine diglyceride transferase; PECT, phosphoethanolamine cytidylyltransferase; SPLY, sphinganine-1-phosphate lyase. *P<0.05, **P<0.01, ****P<0.0001.
- Fig. 7.
Population and temperature acclimation effects on activity (A) and expression (B) of the lipid-mediated signaling enzyme, phospholipase D (PLD). Shown are least square means ± 1 standard error and significant fixed effects from mixed-model ANOVAs. Fold expression changes were calculated from serial dilution standard curves. *P<0.05, ****P<0.0001.
- Fig. 8.
The effect of rapid thermal shifts on ethanol tolerance. Flies reared at 26°C and shifted to 15°C (left) had increased ethanol tolerance, whereas flies reared at 15°C and shifted to 26°C (right) had decreased tolerance. Data are least square means ± 1 standard error.
- Fig. 9.
The effects of a rapid thermal shift from 26°C to 15°C, which should increase membrane rigidity, on the expression of genes involved in phosphatidylethanolamine (PE) biosynthesis (A; Sply, Cdpet), membrane lipid signaling (B; Pld, Wun) and encoding a putative AcCoAS (C; CG6432). Shown are least square means ± 1 standard error and significant fixed effects of TExpose(TRear) from mixed-model ANOVAs. Units of expression are (1/Ct)×1000. Fold expression changes were calculated from serial dilution standard curves. **P<0.01, ***P<0.001, ****P<0.0001.
- Fig. 10.
The response of acetic acid tolerance (A) and dSrebp expression (B) to temperature acclimation. Shown are least square means ± 1 standard error and significant fixed effects. *P<0.05, ****P<0.0001.
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