RESUMO
How species thrive in a wide range of environments is a major focus of evolutionary biology. For many species, limited genetic diversity or gene flow among habitats means that phenotypic plasticity must play an important role in their capacity to tolerate environmental heterogeneity and to colonize new habitats. However, we have a limited understanding of the molecular components that govern plasticity in ecologically relevant phenotypes. We examined this hypothesis in a spider species (Stegodyphus dumicola) with extremely low species-wide genetic diversity that nevertheless occupies a broad range of thermal environments. We determined phenotypic responses to temperature stress in individuals from four climatic zones using common garden acclimation experiments to disentangle phenotypic plasticity from genetic adaptations. Simultaneously, we created data sets on multiple molecular modalities: the genome, the transcriptome, the methylome, the metabolome and the bacterial microbiome to determine associations with phenotypic responses. Analyses of phenotypic and molecular associations reveal that acclimation responses in the transcriptome and metabolome correlate with patterns of phenotypic plasticity in temperature tolerance. Surprisingly, genes whose expression seemed to be involved in plasticity in temperature tolerance were generally highly methylated contradicting the idea that DNA methylation stabilizes gene expression. This suggests that the function of DNA methylation in invertebrates varies not only among species but also among genes. The bacterial microbiome was stable across the acclimation period; combined with our previous demonstrations that the microbiome is temporally stable in wild populations, this is convincing evidence that the microbiome does not facilitate plasticity in temperature tolerance. Our results suggest that population-specific variation in temperature tolerance among acclimation temperatures appears to result from the evolution of plasticity in mainly gene expression.
RESUMO
Plastic responses to heat stress have been shown to temporarily increase heat stress tolerance in many small ectotherms. Heat shock proteins (Hsps) have previously been shown to play a role in this induced heat stress tolerance. The heat shock response is fast but short lived, with the cellular Hsp concentration peaking within a few hours after induction. Induced heat stress tolerance, on the other hand, peaks 16-32 h after induction. Therefore, the inducible heat stress response must depend on additional mechanisms. The Turandot gene family has been suggested as a candidate. It contains eight genes that are all upregulated to some degree following heat stress in Drosophila melanogaster. Previously, Turandot A (totA) and Turandot X (totX) have been linked to induced heat stress tolerance. The study presented here aimed to investigate the temporal dynamics of Turandot expression and the functional role of totA and totC for heat stress tolerance. This was done by assaying the temporal heat tolerance and Turandot gene expression after a heat insult, and by exposing Turandot gene knock down flies to a range of heat hardening treatments, and evaluating the effects on heat tolerance. Successful gene knock down was verified by gene expression assays. In addition, expression of hsp70A was included. Both totA, totC, and hsp70A expression increased following a heat hardening treatment, while the results for totX were less clear. The expression of totC temporally co-occurred with and was functionally linked to increased heat tolerance. Expression of totA did not have a significant effect on heat stress tolerance. The complexity of inducible heat tolerance was underlined by the result that knock down of Turandot genes led to increased expression of hsp70. The results suggest that heat tolerance is determined by the interaction between several mechanisms, of which Turandot genes constitute one such mechanism.
