Description: 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.

Description: Three hundred and fifty specimens of the endemic Southern Ocean octopus genus Pareledone, were sequenced for the barcoding gene COI. Geographic coverage comprised the South Shetland Islands, the Ross Sea, Adélie Land, George V Land, the Weddell Sea, under the site of the former Larsen B ice shelf, Prydz Bay, the South Orkney Islands and the Amundsen Sea. The greatest number of specimens was captured at the three first-mentioned localities. At least 11 species were represented in the samples and the analyses revealed cryptic species. Six species were found to have extended distributions. Circumpolarity is supported for at least one species. Evidence is presented for a barrier to gene flow to the west of the Antarctic Peninsula, with haplotypes of P. aequipapillae becoming progressively more diverse in a clockwise direction from the South Shetland Islands to the Amundsen Sea. This pattern is akin to that seen in ring species, although we suggest that comparatively warm bottom water acts as a physical barrier preventing completion of the ring.

Description: A morphological dataset based on 14 standard counts and indices was constructed for 68 specimens comprising 12 species of octopuses. This was used to construct distance matrices based on morphological characters. These matrices were compared with genetic distance matrices compiled during molecular phylogenetic analyses of the same 12 species using four mitochondrial and two nuclear genes. Mantel tests showed that there was significant congruence between the phenetic and genetic matrices, suggesting that the genetic signal is reflected in the morphological data set. Matrices of geographical distance were constructed for the 12 species based on the latitude, longitude, and depth of capture of 1726 individuals. These matrices never showed significant congruence with genetic data or with morphological data. Multivariate analysis of the morphological dataset suggests that these counts and indices, traditionally used for discriminating between species in cephalopods, do not show great discrimination at species level, but provide excellent discrimination at the generic level, and, as such, might be useful for resolving the generic placement of some problematic taxa.

Description: A new genus of octopus, Sasakiopus, is erected for the species S. salebrosus (Sasaki, 1920) n. comb. Sasakiopus salebrosus is redescribed from the holotype and from new material recently collected in the eastern Bering Sea. Molecular phylogenetic analysis of one nuclear and three mitochondrial genes revealed that the new genus is the sister taxon of a clade containing the genera Benthoctopus and Vulcanoctopus. The clade containing Sasakiopus, Benthoctopus and Vulcanoctopus is the sister group of Enteroctopus. The genus Bathypolypus falls outside this clade. Sasakiopus differs from Bathypolypus and Enteroctopus by the shape of its ligula (simple in Sasakiopus and Benthoctopus, laminate in Bathypolypus and elongate in Enteroctopus), from Enteroctopus by the absence of enlarged suckers in mature male animals and from Benthoctopus by its skin sculpture and ability to ink.

Description: Background: The giant squid (Architeuthis dux; Steenstrup, 1857) is an enigmatic giant mollusc with a circumglobal distribution in the deep ocean, except in the high Arctic and Antarctic waters. The elusiveness of the species makes it difficult to study. Thus, having a genome assembled for this deep-sea-dwelling species will allow several pending evolutionary questions to be unlocked. Findings: We present a draft genome assembly that includes 200 Gb of Illumina reads, 4 Gb of Moleculo synthetic long reads, and 108 Gb of Chicago libraries, with a final size matching the estimated genome size of 2.7 Gb, and a scaffold N50 of 4.8 Mb. We also present an alternative assembly including 27 Gb raw reads generated using the Pacific Biosciences platform. In addition, we sequenced the proteome of the same individual and RNA from 3 different tissue types from 3 other species of squid (Onychoteuthis banksii, Dosidicus gigas, and Sthenoteuthis oualaniensis) to assist genome annotation. We annotated 33,406 protein-coding genes supported by evidence, and the genome completeness estimated by BUSCO reached 92%. Repetitive regions cover 49.17% of the genome. Conclusions: This annotated draft genome of A. dux provides a critical resource to investigate the unique traits of this species, including its gigantism and key adaptations to deep-sea environments.

Description: It is now recognised that the biology of almost any organism cannot be fully understood without recognising the existence and potential functional importance of associated microbes. Arguably, the emergence of this holistic viewpoint may never have occurred without the development of a crucial molecular technique, 16S rDNA amplicon sequencing, which allowed microbial communities to be easily profiled across a broad range of contexts. A diverse array of molecular techniques are now used to profile microbial communities, infer their evolutionary histories, visualise them in host tissues, and measure their molecular activity. In this review, we examine each of these categories of measurement and inference with a focus on the questions they make tractable, and the degree to which their capabilities and limitations shape our view of the holobiont.

Description: The concept of a Barcode of Life Database (BoLD) for the Class Cephalopoda (Phylum Mollusca) was introduced at the Cephalopod International Advisory Council (CIAC) symposium in Hobart, Australia, February 2006. This suggestion was met with significant interest, concern and debate. This review attempts to describe the concept of the BoLD initiative and to outline considerations and concerns specific to a cephalopod BoLD.

Description: To determine the pattern of spatial genetic structure in the endemic Southern Ocean octopus Adelieledone polymorpha, microsatellite loci were isolated from partial genomic libraries enriched for repetitive DNA motifs. Seven dinucleotide and two trinucleotide microsatellite loci were isolated successfully and levels of polymorphism were quantified in 34 individuals sampled from the Southern Ocean near South Georgia. No pairs of microsatellite loci were linked significantly; however, one locus deviated (P 〈 0.05) from Hardy–Weinberg equilibrium. Overall, the nine loci produced between five and 16 alleles, with observed and expected heterozygosities varying between 0.22 and 0.86 and between 0.21 and 0.94 respectively. This is the first description of microsatellite loci from an octopus endemic to the Southern Ocean, and these genetic markers are being used to quantify spatial structure within A. polymorpha.

Description: Understanding how environmental forcing has generated and maintained large-scale patterns of biodiversity is a key goal of evolutionary research and critical to predicting the impacts of global climate change. We suggest that the initiation of the global thermohaline circulation provided a mechanism for the radiation of Southern Ocean fauna into the deep sea. We test this hypothesis using a relaxed phylogenetic approach to coestimate phylogeny and divergence times for a lineage of octopuses with Antarctic and deep-sea representatives. We show that the deep-sea lineage had their evolutionary origins in Antarctica, and estimate that this lineage diverged around 33 million years ago (Ma) and subsequently radiated at 15 Ma. Both of these dates are critical in development of the thermohaline circulation and we suggest that this has acted as an evolutionary driver enabling the Southern Ocean to become a centre of origin for deep-sea fauna. This is the first unequivocal molecular evidence that deep-sea fauna from other ocean basins originated from Southern Ocean taxa and this is the first evidence to be dated.

Description: In the Southern Ocean, that is areas south of the Polar Front, long-term oceanographic cooling, geographic separation, development of isolating current and wind systems, tectonic drift and fluctuation of ice sheets amongst others have resulted in a highly endemic benthic fauna, which is generally adapted to the long-lasting, relatively stable environmental conditions. The Southern Ocean benthic ecosystem has been subject to minimal direct anthropogenic impact (compared to elsewhere) and thus presents unique opportunities to study biodiversity and its responses to environmental change. Since the beginning of the century, research under the Census of Marine Life and International Polar Year initiatives, as well as Scientific Committee of Antarctic Research biology programmes, have considerably advanced our understanding of the Southern Ocean benthos. In this paper, we evaluate recent progress in Southern Ocean benthic research and identify priorities for future research. Intense efforts to sample and describe the benthic fauna, coupled with coordination of information in global databases, have greatly enhanced understanding of the biodiversity and biogeography of the region. Some habitats, such as chemosynthetic systems, have been sampled for the first time, while application of new technologies and methods are yielding new insights into ecosystem structure and function. These advances have also highlighted important research gaps, notably the likely consequences of climate change. In a time of potentially pivotal environmental change, one of the greatest challenges is to balance conservation with increasing demands on the Southern Ocean’s natural resources and services. In this context, the characterization of Southern Ocean biodiversity is an urgent priority requiring timely and accurate species identifications, application of standardized sampling and reporting procedures, as well as cooperation between disciplines and nations.