DNA alignment and Bayesian trees of partial sequences of COI, COIII and the haemocyanin functional unit f-g of 28 octopod species

Background: Octopods have successfully colonised the world's oceans from the tropics to the poles. Yet, successful persistence in these habitats has required adaptations of their advanced physiological apparatus to compensate impaired oxygen supply. Their oxygen transporter haemocyanin plays a major role in cold tolerance and accordingly has undergone functional modifications to sustain oxygen release at sub-zero temperatures. However, it remains unknown how molecular properties evolved to explain the observed functional adaptations. We thus aimed to assess whether natural selection affected molecular and structural properties of haemocyanin that explains temperature adaptation in octopods. Results: Analysis of 239 partial sequences of the haemocyanin functional units (FU) f and g of 28 octopod species of polar, temperate, subtropical and tropical origin revealed natural selection was acting primarily on charge properties of surface residues. Polar octopods contained haemocyanins with higher net surface charge due to decreased glutamic acid content and higher numbers of basic amino acids. Within the analysed partial sequences, positive selection was present at site 2545, positioned between the active copper binding centre and the FU g surface. At this site, methionine was the dominant amino acid in polar octopods and leucine was dominant in tropical octopods. Sites directly involved in oxygen binding or quaternary interactions were highly conserved within the analysed sequence. Conclusions: This study has provided the first insight into molecular and structural mechanisms that have enabled octopods to sustain oxygen supply from polar to tropical conditions. Our findings imply modulation of oxygen binding via charge-charge interaction at the protein surface, which stabilize quaternary interactions among functional units to reduce detrimental effects of high pH on venous oxygen release. Of the observed partial haemocyanin sequence, residue 2545 formed a close link between the FU g surface and the active centre, suggesting a role as allosteric binding site. The prevalence of methionine at this site in polar octopods, implies regulation of oxygen affinity via increased sensitivity to allosteric metal binding. High sequence conservation of sites directly involved in oxygen binding indicates that functional modifications of octopod haemocyanin rather occur via more subtle mechanisms, as observed in this study.

Potential connections (least-cost paths) between existing and potential bison habitat patches in the Carpathian Mountains

Habitat connectivity is important for the survival of species that occupy habitat patches too small to sustain an isolated population. A prominent example of such a species is the European bison (Bison bonasus), occurring only in small, isolated herds, and whose survival will depend on establishing larger, well-connected populations. Our goal here was to assess habitat connectivity of European bison in the Carpathians. We used an existing bison habitat suitability map and data on dispersal barriers to derive cost surfaces, representing the ability of bison to move across the landscape, and to delineate potential connections (as least-cost paths) between currently occupied and potential habitat patches. Graph theory tools were then employed to evaluate the connectivity of all potential habitat patches and their relative importance in the network. Our analysis showed that existing bison herds in Ukraine are isolated. However, we identified several groups of well-connected habitat patches in the Carpathians which could host a large population of European bison. Our analysis also located important dispersal corridors connecting existing herds, and several promising locations for future reintroductions (especially in the Eastern Carpathians) that should have a high priority for conservation efforts. In general, our approach indicates the most important elements within a landscape mosaic for providing and maintaining the overall connectivity of different habitat networks and thus offers a robust and powerful tool for conservation planning.