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Simple Summary The atomic structure of a biological macromolecule determines its function. Discovering how one or more amino acid chains fold and interact to form a protein complex is critical, from understanding the most fundamental cellular processes to developing new therapies. However, this is far from a straightforward task, especially when studying a membrane protein. The functional link between the oligomeric state and complex composition of the proteins involved in the active mitochondrial transport of cytosolic pyruvate is a decades-old question but remains urgent. We present a brief historical review beginning with the identification of the so-called mitochondrial pyruvate carrier (MPC) proteins, followed by a rigorous conceptual analysis of technical approaches in more recent biochemical studies that seek to isolate and reconstitute the functional MPC complex(es) in vitro. We correlate these studies with early kinetic observations and current experimental and computational knowledge to assess their main contributions, identify gaps, resolve ambiguities, and better define the research goal. Abstract The molecular identity of the mitochondrial pyruvate carrier (MPC) was presented in 2012, forty years after the active transport of cytosolic pyruvate into the mitochondrial matrix was first demonstrated. An impressive amount of in vivo and in vitro studies has since revealed an unexpected interplay between one, two, or even three protein subunits defining different functional MPC assemblies in a metabolic-specific context. These have clear implications in cell homeostasis and disease, and on the development of future therapies. Despite intensive efforts by different research groups using state-of-the-art computational tools and experimental techniques, MPCs’ structure-based mechanism remains elusive. Here, we review the current state of knowledge concerning MPCs’ molecular structures by examining both earlier and recent studies and presenting novel data to identify the regulatory, structural, and core transport activities to each of the known MPC subunits. We also discuss the potential application of cryogenic electron microscopy (cryo-EM) studies of MPC reconstituted into nanodiscs of synthetic copolymers for solving human MPC2.

The E. coli membrane protein ZipA, binds to the tubulin homologue FtsZ, in the early stage of cell division. We isolated ZipA in a Styrene Maleic Acid lipid particle (SMALP) preserving its position and integrity with native E. coli membrane lipids. Direct binding of ZipA to FtsZ is demonstrated, including FtsZ fibre bundles decorated with ZipA. Using Cryo-Electron Microscopy, small-angle X-ray and neutron scattering, we determine the encapsulated-ZipA structure in isolation, and in complex with FtsZ to a resolution of 1.6 nm. Three regions can be identified from the structure which correspond to, SMALP encapsulated membrane and ZipA transmembrane helix, a separate short compact tether, and ZipA globular head which binds FtsZ. The complex extends 12 nm from the membrane in a compact structure, supported by mesoscale modelling techniques, measuring the movement and stiffness of the regions within ZipA provides molecular scale analysis and visualisation of the early divisome.

Most membrane proteins function through interactions with other proteins in the phospholipid bilayer, the cytosol or the extracellular milieu. Understanding the molecular basis of these interactions is key to understanding membrane protein function and dysfunction. Here we demonstrate for the first time how a nano-encapsulation method based on styrene maleic acid lipid particles (SMALPs) can be used in combination with native gel electrophoresis to separate membrane protein complexes in their native state. Using four model proteins, we show that this separation method provides an excellent measure of protein quaternary structure, and that the lipid environment surrounding the protein(s) can be probed using mass spectrometry. We also show that the method is complementary to immunoblotting. Finally we show that intact membrane protein-SMALPs extracted from a band on a gel could be visualised using electron microscopy (EM). Taken together these results provide a novel and elegant method for investigating membrane protein complexes in a native state.

Biological characterisation of membrane proteins lags behind that of soluble proteins. This reflects issues with the traditional use of detergents for extraction, as the surrounding lipids are generally lost, with adverse structural and functional consequences. In contrast, styrene maleic acid (SMA) copolymers offer a detergent-free method for biological membrane solubilisation to produce SMA-lipid particles (SMALPs) containing membrane proteins together with their surrounding lipid environment. We report the development of a reverse-phase LC-MS/MS method for bacterial phospholipids and the first comparison of the profiles of SMALP co-extracted phospholipids from three exemplar bacterial membrane proteins with different topographies: FtsA (associated membrane protein), ZipA (single transmembrane helix), and PgpB (integral membrane protein). The data showed that while SMA treatment per se did not preferentially extract specific phospholipids from the membrane, SMALP-extracted ZipA showed an enrichment in phosphatidylethanolamines and depletion in cardiolipins compared to the bulk membrane lipid. Comparison of the phospholipid profiles of the 3 SMALP-extracted proteins revealed distinct lipid compositions for each protein: ZipA and PgpB were similar, but in FtsA samples longer chain phosphatidylglycerols and phosphatidylethanolamines were more abundant. This method offers novel information on the phospholipid interactions of these membrane proteins.

The use of styrene maleic acid lipid particles (SMALPs) for the purification of membrane proteins (MPs) is a rapidly developing technology. The amphiphilic copolymer of styrene and maleic acid (SMA) disrupts biological membranes and can extract membrane proteins in nanodiscs of approximately 10 nm diameter. These discs contain SMA, protein and membrane lipids. There is evidence that MPs in SMALPs retain their native structures and functions, in some cases with enhanced thermal stability. In addition, the method is compatible with biological buffers and a wide variety of biophysical and structural analysis techniques. The use of SMALPs to solubilize and stabilize MPs offers a new approach in our attempts to understand, and influence, the structure and function of MPs and biological membranes. In this review, we critically assess progress with this method, address some of the associated technical challenges, and discuss opportunities for exploiting SMA and SMALPs to expand our understanding of MP biology.

New technologies for the purification of stable membrane proteins have emerged in recent years, in particular methods that allow the preparation of membrane proteins with their native lipid environment. Here, we look at the progress achieved with the use of styrene-maleic acid copolymers (SMA) which are able to insert into biological membranes forming nanoparticles containing membrane proteins and lipids. This technology can be applied to membrane proteins from any host source, and, uniquely, allows purification without the protein ever being removed from a lipid bilayer. Not only do these SMA lipid particles (SMALPs) stabilise membrane proteins, allowing structural and functional studies, but they also offer opportunities to understand the local lipid environment of the host membrane. With any new or different method, questions inevitably arise about the integrity of the protein purified: does it retain its activity; its native structure; and ability to perform its function? How do membrane proteins within SMALPS perform in existing assays and lend themselves to analysis by established methods? We outline here recent work on the structure and function of membrane proteins that have been encapsulated like this in a polymer-bound lipid bilayer, and the potential for the future with this approach. This article is part of a Special Issue entitled: Beyond the Structure-Function Horizon of Membrane Proteins edited by Ute Hellmich, Rupak Doshi and Benjamin McIlwain.

Over the past 50years there has been considerable progress in our understanding of biomolecular interactions at an atomic level. This in turn has allowed molecular simulation methods employing full atomistic modelling at ever larger scales to develop. However, some challenging areas still remain where there is either a lack of atomic resolution structures or where the simulation system is inherently complex. An area where both challenges are present is that of membranes containing membrane proteins. In this review we analyse a new practical approach to membrane protein study that offers a potential new route to high resolution structures and the possibility to simplify simulations. These new approaches collectively recognise that preservation of the interaction between the membrane protein and the lipid bilayer is often essential to maintain structure and function. The new methods preserve these interactions by producing nano-scale disc shaped particles that include bilayer and the chosen protein. Currently two approaches lead in this area: the MSP system that relies on peptides to stabilise the discs, and SMALPs where an amphipathic styrene maleic acid copolymer is used. Both methods greatly enable protein production and hence have the potential to accelerate atomic resolution structure determination as well as providing a simplified format for simulations of membrane protein dynamics. This article is part of a Special Issue entitled: Biosimulations edited by Ilpo Vattulainen and Tomasz Róg.

Protein secretion and membrane insertion occur through the ubiquitous Sec machinery. In this system, insertion involves the targeting of translating ribosomes via the signal recognition particle and its cognate receptor to the SecY (bacteria and archaea)/Sec61 (eukaryotes) translocon. A common mechanism then guides nascent transmembrane helices (TMHs) through the Sec complex, mediated by associated membrane insertion factors. In bacteria, the membrane protein ‘insertase’ YidC ushers TMHs through a lateral gate of SecY to the bilayer. YidC is also thought to incorporate proteins into the membrane independently of SecYEG. Here, we show the bacterial holo-translocon (HTL) — a supercomplex of SecYEG–SecDF–YajC–YidC — is a bona fide resident of the Escherichia coli inner membrane. Moreover, when compared with SecYEG and YidC alone, the HTL is more effective at the insertion and assembly of a wide range of membrane protein substrates, including those hitherto thought to require only YidC.

Despite the great importance of membrane proteins, structural and functional studies of these proteins present major challenges. A significant hurdle is the extraction of the functional protein from its natural lipid membrane. Traditionally achieved with detergents, purification procedures can be costly and time consuming. A critical flaw with detergent approaches is the removal of the protein from the native lipid environment required to maintain functionally stable protein. This protocol describes the preparation of styrene maleic acid (SMA) co-polymer to extract membrane proteins from prokaryotic and eukaryotic expression systems. Successful isolation of membrane proteins into SMA lipid particles (SMALPs) allows the proteins to remain with native lipid, surrounded by SMA. We detail procedures for obtaining 25 g of SMA (4 d); explain the preparation of protein-containing SMALPs using membranes isolated from Escherichia coli (2 d) and control protein-free SMALPS using E. coli polar lipid extract (1-2 h); investigate SMALP protein purity by SDS-PAGE analysis and estimate protein concentration (4 h); and detail biophysical methods such as circular dichroism (CD) spectroscopy and sedimentation velocity analytical ultracentrifugation (svAUC) to undertake initial structural studies to characterize SMALPs (∼2 d). Together, these methods provide a practical tool kit for those wanting to use SMALPs to study membrane proteins.

RPP13, a member of the cytoplasmic class of disease resistance genes, encodes one of the most variable Arabidopsis proteins so far identified. This variability is matched in ATR13, the protein from the oomycete downy mildew pathogen Hyaloperonospora parasitica recognized by RPP13, suggesting that these proteins are involved in tight reciprocal coevolution. ATR13 exhibits five domains: an N-terminal signal peptide, an RXLR motif, a heptad leucine/isoleucine repeat, an 11-amino-acid repeated sequence and a C-terminal domain. We show that the conserved RXLR-containing domain is dispensable for ATR13-mediated recognition, consistent with its role in transport into the plant cytoplasm. Sequencing ATR13 from 16 isolates of H. parasitica revealed high levels of amino acid diversity across the entire protein. The leucines/isoleucines of the heptad leucine repeat were conserved, and mutation of particular leucine or isoleucine residues altered recognition by RPP13. Natural variation has not exploited this route to detection avoidance, suggesting a key role of this domain in pathogenicity. The extensive variation in the 11-amino-acid repeat units did not affect RPP13 recognition. Domain swap analysis showed that recognition specificity lay in the C-terminal domain of ATR13. Variation analyses combined with functional assays allowed the identification of four amino acid positions that may play a role in recognition specificity. Site-directed mutagenesis confirmed that a threonine residue is absolutely required for RPP13 recognition and that recognition can be modulated by the presence of either an arginine or glutamic acid at other sites. Mutations in these three amino acids had no effect on the interaction of ATR13 with a resistance gene unlinked to RPP13, consistent with their critical role in determining RPP13-Nd recognition specificity.

Here we examine the role of competitive interactions in controlling distributions of the most abundant omnivore–detritivore snails in East Coast U.S. salt marshes (Melampus bidentatus and Littoraria irrorata). Both snails prefer to eat fungi growing on plant material, and the periwinkle Littoraria, a much larger snail, destroys marsh canopy when grazing fungal-infected plants, resulting in increased local desiccation stress. To test if Littoraria displaces Melampus from middle marsh habitats (i.e., short Spartina alterniflora zone), we manipulated Littoraria density and shading intensity in a two-year field experiment. Both removing snails and increasing shade dramatically increased Melampus densities. Addition of periwinkles to high marsh plant zones (Juncus roemerianus) where Melampus are naturally common induced significant Melampus migration. Growth experiments under controlled canopy conditions in the middle marsh showed that periwinkle presence suppresses Melampus growth. Experimental results suggest that Littoraria may limit the lower distribution of Melampus directly (through competition) and indirectly (through habitat modification). Lab experiments show that Melampus actively avoid mucus from Littoraria and that Littoraria may dislodge or consume Melampus egg masses. This work identifies several mechanisms of displacement and suggests that periwinkles play an important regulatory role in this detritivore assemblage.

Influential research in terrestrial habitats indicates that several ecosystem processes are related to plant biodiversity, yet these links remain poorly studied in marine ecosystems. We conducted one field and one mesocosm experiment to quantify the relative effects of macroalgal species identity and richness on primary production in coral reef macroalgal communities off the north coast of Jamaica. We measured production as the net accumulation of algal biomass in the absence of consumers and as photosynthetic rate using oxygen probes in sealed aquaria. We used two recently developed techniques to attribute deviations in expected relative yield to components associated with species identity or diversity and then to further partition diversity effects into mechanistic components based on dominance, trait-dependent complementarity, and trait-independent complementarity. Our results indicate that algal identity had far greater effects on absolute net growth and photosynthesis than richness. The most diverse mixture of macroalgae did not outperform the most productive monoculture or the average monoculture in either measure of primary production (i.e. we did not find evidence of either transgressive or non-transgressive overyielding). Trait-independent complementarity effects were positive but dominance and trait-dependent complementarity were both negative and became stronger when richness was increased. Thus the potentially positive influence of species interactions and niche partitioning on production were negated by dominance and other negative selection effects. These results demonstrate that the counteracting influence of component effects can diminish the net richness effects on production. This could explain frequently observed weak net richness effects in other aquatic and terrestrial systems and suggests that life history tradeoffs greatly reduce the potential for ecologically relevant plant biodiversity effects on ecosystem properties.

Theoretical and empirical evidence suggest that positive feedbacks can increase resilience in ecological communities. On Caribbean coral reefs, there have been striking shifts from physically complex communities with high coral cover to relatively homogenous communities dominated by macroalgae, which have persisted for decades. However, little is known about positive feedbacks that may maintain coral reef community states. Here, I explore a potential consumer-mediated feedback on a Jamaican reef by examining how grazing by a keystone herbivore (Diadema antillarum) is enhanced by physical structure, which offer refugia from predation. Surveys revealed that habitat complexity and Diadema density were positively related. Increasing habitat complexity by adding physical structure significantly decreased macroalgal cover and increased the proportion of urchins in algal habitats in field manipulations. Experimental increases in urchin density also decreased macroalgal cover, but did not affect the proportion of urchins in algal habitats. These results suggest that the low habitat complexity of macroalgal-dominated reefs may inhibit an urchin-mediated shift to coral dominance and that positive feedbacks must be considered in reef restoration efforts.

Plant biodiversity can enhance primary production in terrestrial ecosystems, but biodiversity effects are largely unstudied in the ocean. We conducted a series of field and mesocosm experiments to measure the relative effects of macroalgal identity and richness on primary productivity (net photosynthetic rate) and biomass accumulation in hard substratum subtidal communities in North Carolina, USA. Algal identity consistently and strongly affected production; species richness effects, although often significent, were subtle. Partitioning of the net biodiversity effect indicated that complementarity effects were always positive and species were usually more productive in mixtures than in monoculture. Surprisingly, slow growing species performed relatively better in the most diverse treatments than the most productive species, thus selection effects were consistently negative. Our results suggest that several basic mechanisms underlying terrestrial plant biodiversity effects also operate in algal-based marine ecosystems, and thus may be general.

Many Caribbean reefs have experienced a phase-shift in community structure, the principle features being a decline in coral cover and an increase in macroalgal biomass. However, one Jamaican reef—Dairy Bull on the north shore near Discovery Bay—is once again dominated by scleractinian corals and several key species have returned. Living coral cover at 6–8 m depth at Dairy Bull has doubled over the past 9 years and is now ~54%. The absolute cover of Acropora cervicornis was

Mushroom corals endemic to the Indo-Pacific were collected and brought to Discovery Bay, Jamaica, and released on the fore-reef in the mid to late 1960’s by Thomas F. Goreau, an early pioneer in coral research (cf. Goreau et al. 1969 p 180; R. K. Trench, personal communication). Subsequently, the potential problems arising from introduction of an exotic species to the Caribbean were realized. Several attempts to eradicate these non-Caribbean species were then made between 1970 and 1980 and more than 25 adult individuals were removed (cf. Bush et al. 2004; J. Woodley, personal communication). However, re-discovery of two Fungia scutaria individuals in March of 2003 showed that a remnant population of this particular mushroom coral has persisted for over 35 years in the Caribbean (Bush et al. 2004). Since being introduced to Atlantic waters, these corals have displayed remarkable resilience, enduring two destructive hurricanes (Woodley 1992), environmental degradation resulting in collapse of native coral populations (Hughes 1994), and several bleaching events related to thermal stress (Goreau 1990). Understanding the stability and/or flexibility of coralalgal symbioses is of great interest to those concerned with the long-term effects of climate change on reef building corals (Baker 2003). Whether the original symbiont population of F. scutaria was replaced by one of the many local Symbiodinium spp. found in the Caribbean (LaJeunesse 2002; LaJeunesse et al. 2003) is addressed in this report through use of genetic analyses. Tissue from a F. scutaria collected on the fore-reef (15 m) off the north shore of Jamaica was preserved in 95% ethanol, homogenized, and the DNA isolated as described previously (LaJeunesse et al. 2003). PCRdenaturing gradient gel electrophoresis (DGGE) fingerprinting of the ribosomal internal transcribed spacer region (ITS) 2 was performed and the two lowest and most prominent DNA bands were excised and sequenced (Fig. 1). The genome of this Symbiodinium sp. contains two co-dominant sequence variants of the ITS 2. They appear to reflect a phylogenetic transition from C1 (ancestral) to the C1b (derived) sequence. When the DGGE fingerprints and sequences are compared with data from hundreds of Caribbean and Indo-Pacific host taxa, an identical match is made with a symbiont of Pacific origin, type C1b (Fig. 1). That member of Symbiodinium clade C was recently identified in the coral Pavona varians (collected at 3 m) from the southern Great Barrier Reef (GBR) (LaJeunesse et al. 2003) and in Leptastrea purpurea (15 m) and the organ pipe octocoral, Tubipora musica (15 m), from mid-shelf reefs on the central GBR (LaJeunesse et al., in press). It is also found in P. superficialis (6 m) from the Pacific coast of Panama (A.C. Baker and T.C. LaJeunesse, unpublished data) and appears to possess a wide, albeit uncommon, geographic distribution in the Pacific. The identical match of the ITS 2 fingerprint, C1b, from the Jamaican F. scutaria to samples taken from various Pacific hosts indicates that this Symbiodinium sp. is most probably a product of the Indo-Pacific diversification of clade C. Presently, over 50 genetically distinguishable symbiont types, mostly from Symbiodinium clades B and C, have been classified from extensive host collections at several sites in the Caribbean (LaJeunesse 2002; LaJeunesse, in press). They predominantly have limited geographic, host-specific and/or well-defined depth distributions within the Caribbean Sea. Only two types from clade C, namely C1 (S. goreaui) and C3, are known to also occur within a wide diversity of host taxa T. C. LaJeunesse (&) Department of Plant Biology and Institute of Ecology, University of Georgia, Plant Sciences Building, Athens, GA 30602, USA E-mail: lajeunes@fiu.edu Tel.: +1-305-3481317 Fax: +1-305-3481986

Plant biodiversity can enhance primary production in terrestrial ecosystems, but biodiversity effects are largely unstudied in the ocean. We conducted a series of field and mesocosm experiments to measure the relative effects of macroalgal identity and richness on primary productivity (net photosynthetic rate) and biomass accumulation in hard substratum subtidal communities in North Carolina, USA. Algal identity consistently and strongly affected production; species richness effects, although often significent, were subtle. Partitioning of the net biodiversity effect indicated that complementarity effects were always positive and species were usually more productive in mixtures than in monoculture. Surprisingly, slow growing species performed relatively better in the most diverse treatments than the most productive species, thus selection effects were consistently negative. Our results suggest that several basic mechanisms underlying terrestrial plant biodiversity effects also operate in algal-based marine ecosystems, and thus may be general.

Early theories of species diversity proposed that communities at equilibrium are saturated with species. However, experiments in plant communities suggest that many communities are unsaturated and species richness can be increased by adding propagules of new species. We experimentally tested for community saturation and measured the effects of propagule supply on community structure in a benthic marine system. We manipulated propagule supply (arrival of individuals of numerous species) of mobile grazers in experimental mesocosms over multiple generations and, unlike previous tests, we examined the cascading effects of propagule supply on prey (macroalgae) biomass. We found little evidence for saturation, despite the absence of processes such as disturbance and predation that are thought to alleviate saturation in nature. Increasing propagule supply increased the total number of species and made rare species more abundant. Perhaps surprisingly, given the strong effect of propagule supply on species richness, supply-related changes in body size and composition suggest that competitive interactions remained important. Grazer supply also had strong cascading effects on primary production, possibly because of dietary complementarity modified by territorial behavior. Our results indicate that propagule supply can directly influence the diversity and composition of communities of mobile animals. Furthermore, the supply of consumer propagules can have strong indirect effects on prey and fundamental ecosystem properties.

By removing herbivores and promoting increases in macroalgae, overfishing is thought to indirectly cause coral disease and mortality. We performed three field manipulations to test the general hypothesis that overfishing and the subsequent alteration of coral reef trophic dynamics are a cause of coral epizootics. Specifically, we asked whether the presence of macroalgae can influence within- and among-colony spread rates of Caribbean Yellow Band Disease in Montastraea faveolata. Macroalgae were placed next to infected and healthy, adult and small coral colonies to measure effects on disease spread rate, coral growth and coral survival. Surprisingly, the addition of macroalgae did not affect disease severity or coral fitness. Our results indicate that macroalgae have no effect on the severity and dynamics of Caribbean Yellow Band Disease and that fisheries management alone will not mitigate the effects of this important epizootic.

The interactive effects of changing biodiversity of consumers and their prey are poorly understood but are likely to be important under realistic scenarios of biodiversity loss and gain. We performed two factorial manipulations of macroalgal group (greens, reds, and browns) and herbivore species (amphipods, sea urchin, and fish) composition and richness in outdoor mesocosms simulating a subtidal, hard-substratum estuarine community in North Carolina, U.S.A. In the experiment where grazer richness treatments were substitutive, there were no significant effects of algal or herbivore richness on final algal biomass. However, in the experiment in which grazer treatments were additive (i.e., species-specific densities were held constant across richness treatments), we found strong independent and interactive effects of algal and herbivore richness. Herbivore polycultures reduced algal biomass to a greater degree than the sum of the three herbivore monocultures, indicating that the measured grazer richness effects were not due solely to increased herbivore density in the polycultures. Taking grazer density into account also revealed that increasing algal richness dampened grazer richness effects. Additionally, the effect of algal richness on algal biomass accumulation was far stronger when herbivores were absent, suggesting that grazers can utilize the increased productivity and mask the positive effects of plant biodiversity on primary production. Our results highlight the complex independent and interactive effects of biodiversity between adjacent trophic levels and emphasize the importance of performing biodiversity-ecosystem functioning experiments in a realistic multi-trophic context.

A recently identified class of proteins conferring insecticidal activity to several bacteria within the Enterobacteriaceae family have potential for control of commercially important insect pests. Here, we report the first purification, biophysical characterisation and 3-D structural analysis of one of the toxin components, XptA1, from Xenorhabdus nematophila PMFI296 to a resolution of 23 A. Membrane binding studies indicate that the three-component toxin system has a different mode of action from that of proteins from Bacillus thuringiensis (Bt). Biophysical characterisation of XptA1 suggests a mechanism of action of XptA1 whereby it first binds to the cell membrane forming a structure with a central cavity and forms a complex with its partners XptB1 and XptC1 producing the full insecticidal toxin. The structure of XptA1 is shown by a combination of electron microscopy, ultracentrifugation and circular dichroism spectroscopy to be a 1.15 MDa tetramer with a cage-like structure. Each of the four symmetry-related subunits has three well-defined domains and a longitudinal twist with one end narrower than the other. One third of the residues of XptA1 are alpha-helical and it is suggested the subunits associate partly via an alpha-helical coiled-coil interaction. XptA1 itself shows the same secondary structure at neutral pH and in an alkaline environment up to pH10.5. This pH tolerance indicates that the folded XptA1 can pass through the midgut of Lepidopteran insects susceptible to the insecticidal toxin complex. This implies therefore that its folded structure is important for its biological activity.

Any fish species that appears to be readily available in the marketplace will create an impression among the public that there is a plentiful supply of that fish in the sea, but this may belie the true state of the fisheries' stock. Here we use molecular genetic analysis to show that some three-quarters of the fish sold in the United States as 'red snapper'--the US Food and Drug Administration's legally designated common name for Lutjanus campechanus--belong to another species. Mislabelling to this extent not only defrauds consumers but could also adversely affect estimates of stock size if it influences the reporting of catch data that are used in fisheries management.

Although the independent effects of propagule supply and resource availability on species diversity are widely recognized, there are conflicting predictions of how resource availability affects the strength of propagule supply. To determine how propagule supply and resource availability interact to control species richness, we compared the effects of grazer propagule additions on grazer and algal assemblages across an experimental gradient of algal productivity. Different levels of algal productivity (i.e., replacement rate of grazers' food resource) were maintained by manipulating light. Despite light-driven changes in algal standing biomass and diversity, algal resource availability had no effect on propagule limitation of grazer species richness, nor on grazer community composition. Instead, augmenting grazer propagule supply had equally strong, positive effects on grazer richness at all levels of algal resource availability. Although there was evidence for competition and resource limitation in the high propagule supply/low resource treatment, competition was evidently too weak or the experimental duration was too short to lead to competitive exclusion and changes in species richness, evenness, and composition. Our results highlight potential differences in resource use between sessile producers and mobile grazers. Unlike plants that consume inorganic nutrients and light whose renewal is positive, donor-controlled, and identity-fixed, animals consume dynamic resources that can respond to consumption by increasing growth rates, or going extinct and being replaced by other species, as we observed in the replacement of easily ingested green algae by less preferred red algae. The dual nature of these dynamic resources may consequently weaken the link between resource renewal rates and propagule limitation in animal communities.

Any fish species that appears to be readily available in the marketplace will create an impression among the public that there is a plentiful supply of that fish in the sea, but this may belie the true state of the fisheries' stock. Here we use molecular genetic analysis to show that some three-quarters of the fish sold in the United States as 'red snapper'--the US Food and Drug Administration's legally designated common name for Lutjanus campechanus--belong to another species. Mislabelling to this extent not only defrauds consumers but could also adversely affect estimates of stock size if it influences the reporting of catch data that are used in fisheries management.