Molecular evidence of intertidal habitats selecting for repeated ice-binding protein evolution in invertebrates

ABSTRACTIce-binding proteins (IBPs) have evolved independently in multiple taxonomic groups to improve their survival of sub-zero temperatures. Intertidal invertebrates in temperate and polar regions frequently encounter sub-zero temperatures, yet there is little information on IBPs in these organisms. We hypothesized that there are far more ice-binding proteins than are currently known and that the occurrence of freezing in the intertidal zone selects for these proteins. We compiled a list of genome-sequenced invertebrates across multiple habitats and a list of known IBP sequences and used BLAST to identify a wide array of putative IBPs in those invertebrates. We found that the probability of an invertebrate species having an ice-binding protein was significantly greater in intertidal species as compared to those primarily found in open ocean or freshwater habitats. These intertidal IBPs had high sequence similarity to fish and tick antifreeze glycoproteins and fish type II antifreeze proteins. Previously established classifiers based on machine learning techniques further predicted ice-binding activity in the majority of our newly identified putative IBPs. We investigated the potential evolutionary origin of one putative IBP from the hard-shelled mussel Mytilus coruscus and suggest that it arose through gene duplication and neofunctionalization. We show that IBPs likely readily evolve in response to freezing risk, that there is an array of uncharacterized ice binding proteins and highlight the need for broader laboratory-based surveys of the diversity of ice binding activity across diverse taxonomic and ecological groups.Summary statementIntertidal invertebrates have a disproportionate number of putative ice-binding proteins relative to other habitats. These putative proteins are highly similar to antifreeze glycoproteins and type II antifreeze proteins from fish.

Nonconsumptive Predator Effects on Prey Demography: Recent Advances Using Intertidal Invertebrates

Predators influence prey demography through consumption, but the mere presence of predators may trigger behavioural changes in prey that, if persistent or intense, may also influence prey demography. A tractable system to study such nonconsumptive effects (NCEs) of predators involves intertidal invertebrates. This mini review summarises recent research using barnacles and mussels as prey and dogwhelks as predators. The field manipulation of dogwhelk density revealed that pelagic barnacle larvae avoid benthic settlement near dogwhelks, which limits barnacle recruitment, a relevant outcome because recruitment is the only source of population replenishment for barnacles, as they are sessile. This avoidance behaviour is likely triggered by waterborne dogwhelk cues and may have evolved to limit future predation risk. Increasing densities of barnacle recruits and adults can prevent such NCEs from occurring, seemingly because benthic barnacles attract conspecific larvae through chemical cues. Barnacle recruit density increased with the abundance of coastal phytoplankton (food for barnacle larvae and recruits), so barnacle food supply seems to indirectly limit dogwhelk NCEs. By inhibiting barnacle feeding, dogwhelk cues also limited barnacle growth and reproductive output. Wave action weakens dogwhelk NCEs likely through hydrodynamic influences. Dogwhelk cues also limit mussel recruitment, as mussel larvae also exhibit predator avoidance behaviour. The NCEs on recruitment are weaker for mussels than for barnacles, possibly because mussel larvae can detach themselves after initial settlement, an ability that barnacle larvae lack. Overall, these field experiments provide evidence of predator NCEs on prey demography for coastal marine systems.

Drivers of plasticity in freeze tolerance in the intertidal mussel Mytilus trossulus

ABSTRACTFreezing is an extreme stress to living cells, and so freeze-tolerant animals often accumulate protective molecules (termed cryoprotectants) to prevent the cellular damage caused by freezing. The bay mussel, Mytilus trossulus, is an ecologically important intertidal invertebrate that can survive freezing. Although much is known about the biochemical correlates of freeze tolerance in insects and vertebrates, the cryoprotectants that are used by intertidal invertebrates are not well characterized. Previous work has proposed two possible groups of low-molecular weight cryoprotectants in intertidal invertebrates: osmolytes and anaerobic byproducts. In our study, we examined which group of candidate cryoprotectants correlate with plasticity in freeze tolerance in mussels using 1H NMR metabolomics. We found that the freeze tolerance of M. trossulus varies on a seasonal basis, along an intertidal shore-level gradient, and with changing salinity. Acclimation to increased salinity (30 ppt compared with 15 ppt) increased freeze tolerance, and mussels were significantly more freeze tolerant during the winter. Mussel freeze tolerance also increased with increasing shore level. There was limited evidence that anaerobic byproduct accumulation was associated with increased freeze tolerance. However, osmolyte accumulation was correlated with increased freeze tolerance after high salinity acclimation and in the winter. The concentration of most low molecular weight metabolites did not vary with shore level, indicating that another mechanism is likely responsible for this pattern of variation in freeze tolerance. By identifying osmolytes as a group of molecules that assist in freezing tolerance, we have expanded the known biochemical repertoire of the mechanisms of freeze tolerance.

New record of Anthopleura radians Spano & Häussermann, 2017 (Cnidaria: Actiniaria: Actiniidae) from the Mexican Pacific

As part of a biodiversity study on intertidal invertebrates, several sea anemones from the west coast of the Baja California Peninsula were collected in June 2016. The taxonomic features of four specimens agree well with those of the species Anthopleura radians, a recently described sea anemone from northern Chile. This study is the first published report on this species outside the range of distribution documented in the original description, including new images of A. radians’ external and internal features and complete characterization of their cnidae. Differences between A. radians from other species of the genus reported from the eastern Pacific are also discussed.