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.

Distribution of benthic foraminifera in lower Cretaceous sediments of the Cuvuer Abyssal Plain in the Indian Ocean

Analysis of 66 samples from DSDP Site 263 (Cores 263-4R-4 to 263-29R-4) reveals a unique faunal composition with a predominance of agglutinated taxa, many of them previously unrecorded from any other DSDP and ODP Indian Ocean sites. A total of 66 agglutinated and 31 calcareous taxa are documented and five new species are described: Hippocrepina gracilis n.sp., "Textuluriopsis" elegans n.sp., Aaptotoichus challengeri n.sp., "Gaudryinopsis" pseudobettenstaedti n.sp. and "Gaudryina" cuvierensis n.sp. Three assemblages are recognized based on changes in the composition of dominant taxa and occurrences of stratigraphically important species: (1) a high-diversity Valanginian to Barremian Bulbobaculites-Recurvoides Assemblage (Cores 263-29R to - 18R), comprised of numerous elongate agglutinated forms, rare nodosariids, and variable numbers of tubes and ammodiscids; (2) a moderately diverse Aptian to Albian Rhizammina-Ammodiscus-Glomospira Assemblage (Cores 263-18R to -7R) with highly fluctuating numbers of the nominate taxa and Haplophragmoides, Trochammina, Verneuilinoides spp., and Verneuilina howchini; (3) a very low diversity Albian or younger Assemblage (Cores 263-6R to -4R) containing sparse agglutinated foraminifera, rare nodosariids and rotaliids. We interpret the assemblages as shelf to lower slope and consider them to reflect a deepening palaeobathymetry as the Cuvier margin subsided after the initial breakup of East Gondwana during the Valanginian. Our interpretation is in sharp contrast with initial palaeodepth estimates of less than 100 m, as well as with original chronostratigraphic interpretations based on foraminifera and nannofossils which correlated the base of the recovered interval with the Aptian. The absence of many cosmoplitan forms, despite high diversity suggests strong faunal differentiation in the Austral realm or endemisn within the Cuvier Basin during the Early Cretaceous.