CRISPR-Cas9-Mediated Gene Knockout in a Non-Model Sea Urchin, Heliocidaris crassispina.

Sea urchins have been used as model organisms in developmental biology research and the genomes of several sea urchin species have been sequenced. Recently, genome editing technologies have become available for sea urchins, and methods for gene knockout using the CRISPRCas9 system have been established. Heliocidaris crassispina is an important marine fishery resource with edible gonads. Although H. crassispina has been used as a biological research material, its genome has not yet been published, and it is a non-model sea urchin for molecular biology research. However, as recent advances in genome editing technology have facilitated genome modification in non-model organisms, we applied genome editing using the CRISPR-Cas9 system to H. crassispina. In this study, we targeted genes encoding ETS transcription factor (HcEts) and pigmentation-related polyketide synthase (HcPks1). Gene fragments were isolated using primers designed by inter-specific sequence comparisons within Echinoidea. When Ets gene was targeted using two sgRNAs, one successfully introduced mutations and impaired skeletogenesis. In the Pks1 gene knockout, when two sgRNAs targeting the close vicinity of the site corresponding to the target site that showed 100% mutagenesis efficiency of the Pks1 gene in Hemicentrotus pulcherrimus, mutagenesis was not observed. However, two other sgRNAs targeting distant sites efficiently introduced mutations. In addition, Pks1 knockout H. crassispina exhibited an albino phenotype in the pluteus larvae and adult sea urchins after metamorphosis. This indicates that the CRISPRCas9 system can be used to modify the genome of the non-model sea urchin H. crassispina.

The behavior of sympatric sea urchin species across an ecosystem state gradient.

Background: In temperate macroalgal forests, sea urchins are considered as a keystone species due to their grazing ability. Given their potential to shape benthic communities, we monitored the habitat use by three sympatric sea urchin species and compared their behaviors in a vegetated habitat (VH) and an adjacent isoyake habitat (IH). Methods: We monitored the environmental conditions and sea urchin density along deep and shallow transects of the VH and IH for over a year. The benthic rugosity at both sites were also surveyed. A mark-recapture experiment was conducted on the two most abundant sea urchins, Diadema setosum and Heliocidaris crassispina, to elucidate sea urchin movement patterns and group dynamics. Results: We found that exposure to waves was highest at the VH while the IH was sheltered. The deep IH experienced the least amount of light due to high turbidity. Water temperature patterns were similar across sites. The VH benthic topography was more rugose compared to the smoother and silt-covered IH substate. Peak macroalgal bloom occurred three months earlier in IH, but macroalgae persisted longer at the shallow VH. Among the sympatric sea urchins, H. crassispina was most abundant at the shallow VH and was observed in pits and crevices. The most abundant across IH and in the deep VH was D. setosum, preferring either crevices or free-living, depending on hydrodynamic conditions. The least abundant species was D. savignyi, and most often observed in crevices. Small and medium sea urchins were most often observed at the IH site, whereas larger sea urchins were more likely observed at the VH. The mark-recapture study showed that D. setosum was found to displace further at the IH, and H. crassispina was more sedentary. Additionally, D. setosum was always observed in groups, whereas H. crassispina was always solitary. Discussion: The behaviors of sympatric urchins, Diadema savignyi, D. setosum and H. crassispina, differed in response to changes in the benthic environment and physical conditions. Sea urchin displacement increased when rugosity and wave action were low. Habitat preference shifted to crevices in seasons with high wave action. In general, the mark-recapture experiment showed that sea urchins displaced further at night.

Recent reconfiguration of an ancient developmental gene regulatory network in Heliocidaris sea urchins.

Changes in developmental gene regulatory networks (dGRNs) underlie much of the diversity of life, but the evolutionary mechanisms that operate on regulatory interactions remain poorly understood. Closely related species with extreme phenotypic divergence provide a valuable window into the genetic and molecular basis for changes in dGRNs and their relationship to adaptive changes in organismal traits. Here we analyse genomes, epigenomes and transcriptomes during early development in two Heliocidaris sea urchin species that exhibit highly divergent life histories and in an outgroup species. Positive selection and chromatin accessibility modifications within putative regulatory elements are enriched on the branch leading to the derived life history, particularly near dGRN genes. Single-cell transcriptomes reveal a dramatic delay in cell fate specification in the derived state, which also has far fewer open chromatin regions, especially near conserved cell fate specification genes. Experimentally perturbing key transcription factors reveals profound evolutionary changes to early embryonic patterning events, disrupting regulatory interactions previously conserved for ~225 million years. These results demonstrate that natural selection can rapidly reshape developmental gene expression on a broad scale when selective regimes abruptly change. More broadly, even highly conserved dGRNs and patterning mechanisms in the early embryo remain evolvable under appropriate ecological circumstances.

Full-length transcriptome sequencing of Heliocidaris crassispina using PacBio single-molecule real-time sequencing.

The lack of high-throughput sequencing data makes the research progress of Heliocidaris crassispina slow. Therefore, we used PacBio single-molecule real-time sequencing to generate the first full-length transcriptome. Here, 31,181 isoforms were obtained, with an average length of 2383.20 and a N50 length of 2732 bp. Meanwhile, 764 alternative splicing (AS) events, 5098 long-noncoding RNAs (LncRNAs), 6978 simple sequence repeats (SSRs), and 950 hypothetical transcript factors (TFs) were identified. Moreover, five key innate immune pattern recognition receptors (PRRs), including toll-like receptor (TLR), NACHT domain and leucine-rich repeat (NLR), scavenger receptor cysteine-rich (SRCR), peptidoglycan recognition proteins (PGRP), and gram-negative binding proteins (GNBP), were searched in the transcriptome. In addition, 37 isoforms enriched in KEGG and GO immune systems were also detected. The study provid abundant data support for the current research on H. crassispina.

Effects of marine heatwave conditions across the metamorphic transition to the juvenile sea urchin (Heliocidaris erythrogramma).

For short development species, like the sea urchin Heliocidaris erythrogramma, the entire planktonic duration can be impacted by marine heatwaves (MHW). Developmental thermal tolerance of this species through metamorphosis was investigated over a broad range (7.6-28.0 °C), including temperatures across its distribution and MHW conditions. In controls (19.5-21.0 °C), 80% of individuals developed to metamorphosis at day 5, doubling to 10 days at 14.0 °C. The thermal range (14.4-21.2 °C) of metamorphosis on day 7 reflected the realised thermal niche with 25.9 °C the upper temperature for success (T40). By day 10, juvenile tolerance narrowed to the local range (16.2-19.0 °C), similar to levels tolerated by adults, indicating negative carryover effects across the metamorphic transition. Without phenotypic adjustment or adaptation, regional warming will be detrimental, although populations may be sustained by thermotolerant offspring. Our results show the importance of the metamorphic transition in understanding the cumulative sensitivity of species to MHW.

Ocean acidification induces distinct transcriptomic responses across life history stages of the sea urchin Heliocidaris erythrogramma.

Ocean acidification (OA) from seawater uptake of rising carbon dioxide emissions impairs development in marine invertebrates, particularly in calcifying species. Plasticity in gene expression is thought to mediate many of these physiological effects, but how these responses change across life history stages remains unclear. The abbreviated lecithotrophic development of the sea urchin Heliocidaris erythrogramma provides a valuable opportunity to analyse gene expression responses across a wide range of life history stages, including the benthic, post-metamorphic juvenile. We measured the transcriptional response to OA in H. erythrogramma at three stages of the life cycle (embryo, larva, and juvenile) in a controlled breeding design. The results reveal a broad range of strikingly stage-specific impacts of OA on transcription, including changes in the number and identity of affected genes; the magnitude, sign, and variance of their expression response; and the developmental trajectory of expression. The impact of OA on transcription was notably modest in relation to gene expression changes during unperturbed development and much smaller than genetic contributions from parentage. The latter result suggests that natural populations may provide an extensive genetic reservoir of resilience to OA. Taken together, these results highlight the complexity of the molecular response to OA, its substantial life history stage specificity, and the importance of contextualizing the transcriptional response to pH stress in light of normal development and standing genetic variation to better understand the capacity for marine invertebrates to adapt to OA.

Distant hybrids of Heliocidaris crassispina (♀) and Strongylocentrotus intermedius (♂): identification and mtDNA heteroplasmy analysis.

BACKGROUND: Distant hybridization between the sea urchin Heliocidaris crassispina (♀) and the sea urchin Strongylocentrotus intermedius (♂) was successfully performed under laboratory conditions. A new variety of hybrid sea urchin (HS hybrid) was obtained. However, the early-development success rates for the HS hybrids were significantly lower than those of purebred H. crassispina or S. intermedius offspring. In addition, it was difficult to distinguish the HS-hybrid adults from the pure H. crassispina adults, which might lead to confusion in subsequent breeding attempts. In this study, we attempted to develop a method to quickly and effectively identify HS hybrids, and to preliminarily investigate the molecular mechanisms underlying the poor early-development success rates in the HS hybrids. RESULTS: The hybrid sea urchins (HS hybrids) were identified both morphologically and molecularly. There were no significant differences in the test height to test diameter ratios between the HS hybrids and the parents. The number and arrangement of ambulacral pore pairs in the HS hybrids differed from those of the parental lines, which might serve as a useful morphological character for the identification of the HS hybrids. A primer pair that identified the HS hybrids was screened by comparing the mitochondrial genomes of the parental lines. Moreover, paternal leakage induced mitochondrial DNA heteroplasmy in the HS hybrids, which might explain the low rates of early development success in these hybrids. CONCLUSIONS: The distant-hybrid sea urchins were accurately identified using comparative morphological and molecular genetic methods. The first evidence of mtDNA heteroplasmy after the distant hybridization of an echinoderm was also provided.

Ecological implications of purple sea urchin (Heliocidaris crassispina, Agassiz, 1864) enhancement on the coastal benthic food web: evidence from stable isotope analysis.

A responsible approach to marine stock enhancement is an effective approach to restore fishery resources. While the release strategy of target species has been well investigated, the impacts on local ecological equilibrium and habitat qualities have only been poorly considered. In the present study, we evaluated how the macro-benthic food web in Daya Bay was affected by purple sea urchin (Heliocidaris crassispina (Agassiz, 1864) stock enhancement using stable isotope analyses (δ13C and δ15N). Our results indicated that the distribution of local species and trophic diversity were influenced to a certain degree by release of purple sea urchins and changes in the feeding habit of the urchins were observed in line with food abundance, which seasonally varied. When food is abundant, the main food source of sea urchins was microphytobenthos and no significant differences were observed among sites; significant differences in the diet of purple sea urchins were detected when food is less abundant. These results suggested that optimization of the release strategy should include information on seasonal productivity of local recipient sites, food web structure and feeding habits of released species. Such information is essential for building a responsible release approach to maximize production enhancement.

Arachidonic acid and lysophosphatidylcholine inhibit multiple late steps of regulated exocytosis.

The canonical Phospholipase A2 (PLA2) metabolites lysophosphatidylcholine (LPC) and arachidonic acid (ARA) affect regulated exocytosis in a wide variety of cells and are proposed to directly influence membrane merger owing to their respective spontaneous curvatures. According to the Stalk-pore hypothesis, negative curvature ARA inhibits and promotes bilayer merger upon introduction into the distal or proximal monolayers, respectively; in contrast, with positive curvature, LPC has the opposite effects. Using fully primed, release-ready native cortical secretory vesicles (CV), well-established fusion assays and standardized lipid analyses, we show that exogenous ARA and LPC, as well as their non-metabolizable analogous, ETYA and ET-18-OCH3, inhibit the docking/priming and membrane merger steps, respectively, of regulated exocytosis.

Development of the sea urchin Heliocidaris crassispina from Hong Kong is robust to ocean acidification and copper contamination.

Metallic pollution is of particular concern in coastal cities. In the Asian megacity of Hong Kong, despite water qualities have improved over the past decade, some local zones are still particularly affected and could represent sinks for remobilization of labile toxic species such as copper. Ocean acidification is expected to increase the fraction of the most toxic form of copper (Cu2+) by 2.3-folds by 2100 (pH ≈7.7), increasing its bioavailability to marine organisms. Additionally, multiple stressors are likely to exert concomitant effects (additive, synergic or antagonist) on the organisms living in the sea. Here, we tested the hypothesis that copper-contaminated waters are more toxic to sea urchin larvae under future pH conditions. We exposed sea urchin embryos and larvae to two low-pH and two copper treatments (0.1 and 1.0 µM) in three separate experiments. Over the short time typically used for toxicity tests (up to 4-arm plutei, i.e. 3 days), larvae of the sea urchin Heliocidaris crassispina were robust and survived the copper levels present in Hong Kong waters today (≤0.19 µM) as well as the average pH projected for 2100. We, however, observed significant mortality with lowering pH in the longer, single-stressor experiment (Expt A: 8-arm plutei, i.e. 9 days). Abnormality and arm asymmetry were significantly increased by pH or/and by copper presence (depending on the experiment and copper level). Body size (d3; but not body growth rates in Expt A) was significantly reduced by both lowered pH and added copper. Larval respiration (Expt A) was doubled by a decrease at pHT from 8.0 to 7.3 on d6. In Expt B1.0 and B0.1, larval morphology (relative arm lengths and stomach volume) were affected by at least one of the two investigated factors. Although the larvae appeared robust, these sub-lethal effects may have indirect consequences on feeding, swimming and ultimately survival. The complex relationship between pH and metal speciation/uptake is not well-characterized and further investigations are urgently needed to detangle the mechanisms involved and to identify possible caveats in routinely used toxicity tests.

Future warming and acidification result in multiple ecological impacts to a temperate coralline alga.

Coralline algae are a crucial component of reef systems, stabilising reef substrate, providing habitat and contributing to accretion. Coralline algae and their surface microbial biofilms are also important as settlement cues for marine invertebrates, yet few studies address the impact of future environmental conditions on interactions between coralline algae, reef microbes and settlement by larvae of marine invertebrates. We exposed the temperate coralline algal species Amphiroa gracilis to warming and/or acidification scenarios for 21 days. Algae became bleached but photosystem II function was not measurably impacted. Settlement by larvae of the sea urchin Heliocidaris erythrogramma was reduced and the structure of the prokaryotic community associated with A. gracilis was altered. Coralline algae in ambient conditions were dominated by Alphaproteobacteria from the Rhodobacteraceae including Loktonella; those under warming were dominated by Bacteroidetes and Verrucomicrobia; acidification resulted in less Loktonella and more Planctomycetes and a combination of warming and acidification caused increases in Bacteroidetes, Verrucomicrobia and the Alphaproteobacteria family Hyphomonadaceae. These experiments indicate that predicted future environmental change may reduce the ability of some temperate reef coralline algae and associated reef microbes to facilitate settlement of invertebrate larvae as well as having a direct impact to algae via bleaching.

Gadolinium perturbs expression of skeletogenic genes, calcium uptake and larval development in phylogenetically distant sea urchin species.

Chelates of Gadolinium (Gd), a lanthanide metal, are employed as contrast agents for magnetic resonance imaging and are released into the aquatic environment where they are an emerging contaminant. We studied the effects of environmentally relevant Gd concentrations on the development of two phylogenetically and geographically distant sea urchin species: the Mediterranean Paracentrotus lividus and the Australian Heliocidaris tuberculata. We found a general delay of embryo development at 24h post-fertilization, and a strong inhibition of skeleton growth at 48h. Total Gd and Ca content in the larvae showed a time- and concentration-dependent increase in Gd, in parallel with a reduction in Ca. To investigate the impact of Gd on the expression of genes involved in the regulation of skeletogenesis, we performed comparative RT-PCR analysis and found a misregulation of several genes involved in the skeletogenic and left-right axis specification gene regulatory networks. Species-specific differences in the biomineralization response were evident, likely due to differences in the skeletal framework of the larvae and the amount of biomineral produced. Our results highlight the hazard of Gd for marine organisms.

Nodal and BMP expression during the transition to pentamery in the sea urchin Heliocidaris erythrogramma: insights into patterning the enigmatic echinoderm body plan.

BACKGROUND: The molecular mechanisms underlying the development of the unusual echinoderm pentameral body plan and their likeness to mechanisms underlying the development of the bilateral plans of other deuterostomes are of interest in tracing body plan evolution. In this first study of the spatial expression of genes associated with Nodal and BMP2/4 signalling during the transition to pentamery in sea urchins, we investigate Heliocidaris erythrogramma, a species that provides access to the developing adult rudiment within days of fertilization. RESULTS: BMP2/4, and the putative downstream genes, Six1/2, Eya, Tbx2/3 and Msx were expressed in the earliest morphological manifestation of pentamery during development, the five hydrocoele lobes. The formation of the vestibular ectoderm, the specialized region overlying the left coelom that forms adult ectoderm, involved the expression of putative Nodal target genes Chordin, Gsc and BMP2/4 and putative BMP2/4 target genes Dlx, Msx and Tbx. The expression of Nodal, Lefty and Pitx2 in the right ectoderm, and Pitx2 in the right coelom, was as previously observed in other sea urchins. CONCLUSION: That genes associated with Nodal and BMP2/4 signalling are expressed in the hydrocoele lobes, indicates that they have a role in the developmental transition to pentamery, contributing to our understanding of how the most unusual body plan in the Bilateria may have evolved. We suggest that the Nodal and BMP2/4 signalling cascades might have been duplicated or split during the evolution to pentamery.

Microbulbifer echini sp. nov., isolated from the gastrointestinal tract of a purple sea urchin, Heliocidaris crassispina.

A novel bacterium, designated as strain AM134T, was isolated from the gut of a purple sea urchin (Heliocidaris crassispina) gathered from the coastal waters of Dokdo, Korea. Strain AM134T was Gram-stain-negative, both catalase- and oxidase-positive, strictly aerobic and showed a rod-coccus cell cycle. Optimum growth occurred at 30 °C, in the presence of 2 % (w/v) NaCl and at pH 7. The 16S rRNA gene sequence analysis showed that strain AM134T belonged to the genus Microbulbifer in the family Alteromonadaceae and had high 16S rRNA gene sequence similarity (>97 %) with Microbulbifer epialgicus F-104T (98.9 % similarity) and Microbulbifer variabilis Ni-2088T (98.6 % similarity). The polar lipid profile of strain AM134T was composed of phosphatidylethanolamine, phosphatidylserine, three unidentified aminophospholipids, two unidentified phospholipids, an unidentified amino lipid and six unidentified lipids. The major respiratory quinone was identified as ubiquinone-8 (Q-8). The major cellular fatty acids were summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c) and C16 : 0. The DNA-DNA hybridization analysis showed that the strain shared less than 28 % genomic relatedness with Microbulbifer epialgicus DSM 18651T (27±3 %) and Microbulbifer variabilis ATCC 700307T (15±1 %). The G+C content of the genomic DNA was 56.1 mol%. The results of the phylogenetic, phenotypic and genotypic analyses suggest that strain AM134T represents a novel species in the genus Microbulbifer, for which the name Microbulbifer echini is proposed. The type strain is AM134T (=KACC 18258T=JCM 30400T).

Arthrobacter echini sp. nov., isolated from the gut of a purple sea urchin, Heliocidaris crassispina.

A novel strain, designated AM23T, was isolated from the gut of a purple sea urchin Heliocidaris crassispina collected from the coastal waters of the Korean island Dokdo. 16S rRNA gene sequence analysis showed that strain AM23T belonged to the genus Arthrobacter in the family Micrococcaceae and shared highest sequence similarity with Arthrobacter agilis DSM 20550T (98.77%). Strain AM23T was catalase-positive, oxidase-negative and grew optimally at 20 °C, in the presence of 1% (w/v) NaCl and at pH 7. The isolate was a Gram-stain-positive, non-motile, strictly aerobic and coccus-shaped bacterium. The major cellular fatty acids were anteiso-C15:0 and iso-C15:0. The polar lipids of strain AM23T were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol, one unidentified glycolipid and four unidentified lipids. The components of the cell-wall peptidoglycan were lysine, glutamic acid and alanine and the predominant cell-wall sugars were galactose, mannose, rhamnose and ribose. The major respiratory quinone was identified as menaquinone MK-9(H2). The genomic DNA G+C content was 67.3 mol% and the DNA-DNA hybridization values showed the strain shared less than 29% genomic relatedness with A. agilis DSM 20550T. The results of the phylogenetic, phenotypic and genotypic analysis indicate that strain AM23T represents a novel species in the genus Arthrobacter, for which the name Arthrobacter echini sp. nov. is proposed. The type strain is AM23T (=KACC 18260T=DSM 29493T).

Complete mitochondrial genome sequence of Heliocidaris crassispina (Camarodonta, Echinometridae).

The whole mitochondrial genome sequence of sea urchin Heliocidaris crassispina of the family Echinometridae is determined for the first time in this study. The circular mitogenome (15,702 bp) consists of typical Camarodonta gene order and its components including 2 rRNA, 22 tRNA, 13 protein-coding genes and a control region. Phylogenetic analysis based on the 13 concatenated protein-coding gene sequences shows that H. crassispina is closer to the species of Strongylocentrotidae than Parechinidae, but the separation between H. crassispina and the Strongylocentrotid species occurred early in their evolution. The complete mitochondrial genome presented in this study is useful for inferring the phylogenetic relationship among the families of Echinidea sea urchins.

Transcriptomic analysis of Nodal- and BMP-associated genes during juvenile development of the sea urchin Heliocidaris erythrogramma.

Understanding the unusual radial body plan of echinoderms and its relationship to the bilateral plan of other deuterostomes remains a challenge. The molecular processes of embryonic and early larval development in sea urchins are well characterised, but those giving rise to the adult and its radial body remain poorly studied. We used the developmental transcriptome generated for Heliocidaris erythrogramma, a species that forms the juvenile soon after gastrulation, to investigate changes in gene expression underlying radial body development. As coelomogenesis is key to the development of pentamery and juvenile formation on the left side of the larva, we focussed on genes associated with the nodal and BMP2/4 network that pattern this asymmetry. We identified 46 genes associated with this Nodal and BMP2/4 signalling network, and determined their expression profiles from the gastrula, through to rudiment development, metamorphosis and the fully formed juvenile. Genes associated with Nodal signalling shared similar expression profiles, indicating that they may have a regulatory relationship in patterning morphogenesis of the juvenile sea urchin. Similarly, many genes associated with BMP2/4 signalling had similar expression profiles through juvenile development. Further examination of the roles of Nodal- and BMP2/4-associated genes is required to determine function and whether the gene expression profiles seen in H. erythrogramma are due to ongoing activity of gene networks established during early development, or to redeployment of regulatory cassettes to pattern the adult radial body plan.

Early development of congeneric sea urchins (Heliocidaris) with contrasting life history modes in a warming and high CO2 ocean.

The impacts of ocean change stressors - warming and acidification - on marine invertebrate development have emerged as a significant impact of global change. We investigated the response of early development to the larval stage in sympatric, congeneric sea urchins, Heliocidaris tuberculata and Heliocidaris erythrogramma with contrasting modes of development to ocean warming and acidification. Effects of these stressors were assessed by quantifying the percentage of normal development during the first 24 h post fertilization, in cross-factorial experiments that included three temperature treatments (control: 20 °C; +4: 24 °C; +6: 26 °C) and four pHNIST levels (control: 8.2; -0.4: 7.8; -0.6: 7.6; -0.8: 0.4). The experimental treatments were designed in context with present day and near-future (∼2100) conditions for the southeast Australia global warming hotspot. Temperature was the most important factor affecting development of both species causing faster progression through developmental stages as well as a decrease in the percentage of normal development. H. erythrogramma embryos were less tolerant of increased temperature than those of H. tuberculata. Acidification impaired development to the larval stage in H. tuberculata, but this was not the case for H. erythrogramma. Thus, outcomes for the planktonic life phase of the two Heliocidaris species in response to ocean warming and acidification will differ. As shown for these species, single-stressor temperature or acidification studies can be misleading with respect to determining species' vulnerability and responses to global change.

Determination of toxic heavy metals in Echinodermata and Chordata species from South Korea.

This study aimed at analysing concentrations of heavy metals including arsenic, lead, cadmium, aluminium and mercury in commonly consumed seafood species belonging to Echinodermata (Anthocidaris crassispina and Stichopus japonicus) and Chordata (Halocynthia roretzi and Styela plicata). The samples were digested by a microwave system and analysed for As, Cd and Pb by inductively coupled plasma mass spectrometer, for Al by inductively coupled plasma-optical emission spectrometer and Hg by Direct Mercury Analyser. The analytical method was validated by determining sensitivity, linearity, precision, spiking recoveries and analysis of the Standard Reference Material (SRM) NIST 1566-b, an Oyster Tissue. Results showed considerably higher accumulation of Al and As in analysed samples, compared to Pb and Cd, while Hg had the lowest contamination. On comparison, the obtained results with the recommended standards by the Food and Agriculture Organization, European Commission and Ministry of Food and Drug Safety of Korea, it was concluded that the analysed seafoods were safe and thus would not pose a threat to consumers.

Genetic variation underlies temperature tolerance of embryos in the sea urchin Heliocidaris erythrogramma armigera.

Ocean warming can alter natural selection on marine systems, and in many cases, the long-term persistence of affected populations will depend on genetic adaptation. In this study, we assess the potential for adaptation in the sea urchin Heliocidaris erythrogramma armigera, an Australian endemic, that is experiencing unprecedented increases in ocean temperatures. We used a factorial breeding design to assess the level of heritable variation in larval hatching success at two temperatures. Fertilized eggs from each full-sibling family were tested at 22 °C (current spawning temperature) and 25 °C (upper limit of predicted warming this century). Hatching success was significantly lower at higher temperatures, confirming that ocean warming is likely to exert selection on this life-history stage. Our analyses revealed significant additive genetic variance and genotype-by-environment interactions underlying hatching success. Consistent with prior work, we detected significant nonadditive (sire-by-dam) variance in hatching success, but additionally found that these interactions were modified by temperature. Although these findings suggest the potential for genetic adaptation, any evolutionary responses are likely to be influenced (and possibly constrained) by complex genotype-by-environment and sire-by-dam interactions and will additionally depend on patterns of genetic covariation with other fitness traits.