Geographic variation in acclimation responses of thermal tolerance in South African diving beetles (Dytiscidae: Coleoptera).

Understanding sources of variation in animal thermal limits is critical to forecasting ecological responses to climate change. Here, we estimated upper and lower thermal limits, and their capacity to respond to thermal acclimation, in several species and populations of diving beetles (Dytiscidae) from diverse geographic regions representative of variable climate within South Africa. We also considered ecoregions and latitudinal ranges as potential predictors of thermal limits and the plasticity thereof. For upper thermal limits, species showed significant variation and limited acclimation-related plasticity. Lower thermal limits responded to acclimation in some cases and showed marked variation among species that could be explained by taxonomic affiliation and ecoregion. Limited acclimation ability in the species included in this study suggest plasticity of thermal limits will not be a likely buffer for coping with climate change. From the present results for the Dytiscidae of the region, it appears the group may be particularly susceptible to heat and/or drought and may thus serve as useful indicator species of ecosystem change. Understanding how these climate-related impacts play out at different spatial and temporal scales will have profound implications for conservation management and functional responses, especially important in a region already showing a trend for warming and drying.

Thermal niche evolution and geographical range expansion in a species complex of western Mediterranean diving beetles.

BACKGROUND: Species thermal requirements are one of the principal determinants of their ecology and biogeography, although our understanding of the interplay between these factors is limited by the paucity of integrative empirical studies. Here we use empirically collected thermal tolerance data in combination with molecular phylogenetics/phylogeography and ecological niche modelling to study the evolution of a clade of three western Mediterranean diving beetles, the Agabus brunneus complex. RESULTS: The preferred mitochondrial DNA topology recovered A. ramblae (North Africa, east Iberia and Balearic islands) as paraphyletic, with A. brunneus (widespread in the southwestern Mediterranean) and A. rufulus (Corsica and Sardinia) nested within it, with an estimated origin between 0.60-0.25 Ma. All three species were, however, recovered as monophyletic using nuclear DNA markers. A Bayesian skyline plot suggested demographic expansion in the clade at the onset of the last glacial cycle. The species thermal tolerances differ significantly, with A. brunneus able to tolerate lower temperatures than the other taxa. The climatic niche of the three species also differs, with A. ramblae occupying more arid and seasonal areas, with a higher minimum temperature in the coldest month. The estimated potential distribution for both A. brunneus and A. ramblae was most restricted in the last interglacial, becoming increasingly wider through the last glacial and the Holocene. CONCLUSIONS: The A. brunneus complex diversified in the late Pleistocene, most likely in south Iberia after colonization from Morocco. Insular forms did not differentiate substantially in morphology or ecology, but A. brunneus evolved a wider tolerance to cold, which appeared to have facilitated its geographic expansion. Both A. brunneus and A. ramblae expanded their ranges during the last glacial, although they have not occupied areas beyond their LGM potential distribution except for isolated populations of A. brunneus in France and England. On the islands and possibly Tunisia secondary contact between A. brunneus and A. ramblae or A. rufulus has resulted in introgression. Our work highlights the complex dynamics of speciation and range expansions within southern areas during the last glacial cycle, and points to the often neglected role of North Africa as a source of European biodiversity.

Late Miocene diversification of the genus Hydrochus (Coleoptera, Hydrochidae) in the west Mediterranean area.

We provide a reconstruction of the phylogenetic relationships, the geographical and temporal origin, and the mode of diversification of the Mediterranean species of the aquatic beetle family Hydrochidae (Coleoptera, Hydrophiloidea). A total of ca. 3KB of sequence data of three mitochondrial and two nuclear genes were used to reconstruct the phylogeny of 62 specimens of 21 species of Hydrochus, including all western Mediterranean species but one. We estimated the times of divergence using Bayesian methods and an evolutionary rate of 0.0115 substitutions/site/MY, and used an ultrametric calibrated tree to construct a Lineage Through Time (LTT) plot to test alternative models of diversification. A well resolved, well supported phylogeny showed that all western Mediterranean Hydrochus formed a clade, sister to a group including species with a central and eastern European distribution. The origin of the western Mediterranean clade was estimated to be at ca. 13MY, and the speciation events took place between this time and the end of the Messinian, at about 5.3MY. The LTT plot best fitted a model with a shift in the rate of diversification at ca. 8 MY, with a single speciation event (originating two Iberian endemics) subsequent to this period. We conclude that most of the western Mediterranean species of Hydrochidae, including the Ibero-Maghrebian endemics, are ancient elements likely to have remained in the same geographical area since their Miocene origin. Our results add to a growing body of evidence showing the importance of Mediterranean long-term, Tertiary refugia as both cradles and museums of diversity.

Reproducibility and consistency of proteomic experiments on natural populations of a non-model aquatic insect.

Population proteomics has a great potential to address evolutionary and ecological questions, but its use in wild populations of non-model organisms is hampered by uncontrolled sources of variation. Here we compare the response to temperature extremes of two geographically distant populations of a diving beetle species (Agabus ramblae) using 2-D DIGE. After one week of acclimation in the laboratory under standard conditions, a third of the specimens of each population were placed at either 4 or 27°C for 12 h, with another third left as a control. We then compared the protein expression level of three replicated samples of 2-3 specimens for each treatment. Within each population, variation between replicated samples of the same treatment was always lower than variation between treatments, except for some control samples that retained a wider range of expression levels. The two populations had a similar response, without significant differences in the number of protein spots over- or under-expressed in the pairwise comparisons between treatments. We identified exemplary proteins among those differently expressed between treatments, which proved to be proteins known to be related to thermal response or stress. Overall, our results indicate that specimens collected in the wild are suitable for proteomic analyses, as the additional sources of variation were not enough to mask the consistency and reproducibility of the response to the temperature treatments.