Bacterial catabolism of acetovanillone, a lignin-derived compound.

Bacterial catabolic pathways have considerable potential as industrial biocatalysts for the valorization of lignin, a major component of plant-derived biomass. Here, we describe a pathway responsible for the catabolism of acetovanillone, a major component of several industrial lignin streams. Rhodococcus rhodochrous GD02 was previously isolated for growth on acetovanillone. A high-quality genome sequence of GD02 was generated. Transcriptomic analyses revealed a cluster of eight genes up-regulated during growth on acetovanillone and 4-hydroxyacetophenone, as well as a two-gene cluster up-regulated during growth on acetophenone. Bioinformatic analyses predicted that the hydroxyphenylethanone (Hpe) pathway proceeds via phosphorylation and carboxylation, before ß-elimination yields vanillate from acetovanillone or 4-hydroxybenzoate from 4-hydroxyacetophenone. Consistent with this prediction, the kinase, HpeHI, phosphorylated acetovanillone and 4-hydroxyacetophenone. Furthermore, HpeCBA, a biotin-dependent enzyme, catalyzed the ATP-dependent carboxylation of 4-phospho-acetovanillone but not acetovanillone. The carboxylase's specificity for 4-phospho-acetophenone (kcat/KM = 34 ± 2 mM-1 s-1) was approximately an order of magnitude higher than for 4-phospho-acetovanillone. HpeD catalyzed the efficient dephosphorylation of the carboxylated products. GD02 grew on a preparation of pine lignin produced by oxidative catalytic fractionation, depleting all of the acetovanillone, vanillin, and vanillate. Genomic and metagenomic searches indicated that the Hpe pathway occurs in a relatively small number of bacteria. This study facilitates the design of bacterial strains for biocatalytic applications by identifying a pathway for the degradation of acetovanillone.

Ecological generalism and physiology mediate fish biogeographic ranges under ocean warming.

Climate-driven species redistributions are facilitated by niche modifications that increase a species's chances of establishment in novel communities. It is well understood how range-extending species adjust individual niche traits when entering novel environments, yet whether modification of ecological niche traits collectively alters the pace of range extensions or contractions remains unknown. We quantified habitat niche, abundance, physiological performance and cellular defence/damage of range-extending coral reef fishes and coexisting local temperate fishes along a 2000 km latitudinal gradient. We also assessed their dietary and behavioural niches, and establishment potential, to understand whether ecological generalism facilitates successful range extension of coral reef fishes. The coral reef fish that increased all ecological niches, showed stronger establishment, increased physiological performance and cellular damage, but decreased cellular defence at their cold-range edge, whereas tropical species that showed unmodified ecological niches showed lower establishment. One temperate species showed decreased abundance, habitat niche width and body condition, but increased cellular defence, cellular damage and energy reserves at their warm-trailing range, while other temperate species showed contrasting responses. Therefore, ecological generalists might be more successful than ecological specialists during the initial stages of climate change, with increasing future warming strengthening this pattern by physiologically benefitting tropical generalists but disadvantaging temperate specialists.

Body size mediates trophic interaction strength of novel fish assemblages under climate change.

Ecological similarity plays an important role in biotic interactions. Increased body size similarity of competing species, for example, increases the strength of their biotic interactions. Body sizes of many exothermic species are forecast to be altered under global warming, mediating shifts in existing trophic interactions among species, in particular for species with different thermal niches. Temperate rocky reefs along the southeast coast of Australia are located in a climate warming hotspot and now house a mixture of temperate native fish species and poleward range-extending tropical fishes (vagrants), creating novel species assemblages. Here, we studied the relationship between body size similarity and trophic overlap between individual temperate native and tropical vagrant fishes. Dietary niche overlap between vagrant and native fish species increased as their body sizes converged, based on both stomach content composition (short-term diet), stable isotope analyses (integrated long-term diet) and similarity in consumed prey sizes. We conclude that the warming-induced faster growth rates of tropical range-extending fish species at their cool water ranges will continue to converge their body size towards and strengthen their degree of trophic interactions and dietary overlap with co-occurring native temperate species under increasing ocean warming. The strengthening of these novel competitive interactions is likely to drive changes to temperate food web structures and reshuffle existing species community structures.

Staining protocols affect use of otolith to estimate the demography of the damselfish sergeant major (Abudefduf vaigiensis).

This study assessed the otolith (sagittae, lapilli, and asterisci) increment deposition rate in the range-shifting damselfish, A. vaigiensis, using different concentrations of Alizain Red S and evaluated the impact of staining on increment width. Daily increment deposition was verified in all otolith types and presented clearer fluorescent markings in the lapilli and sagittae than the asterisci, with high stain concentration showing the best clarity. Higher stain concentrations were found to decrease increment width, suggesting care is needed when using stained otoliths as a proxy for growth for this species.

Characterization of alkylguaiacol-degrading cytochromes P450 for the biocatalytic valorization of lignin.

Cytochrome P450 enzymes have tremendous potential as industrial biocatalysts, including in biological lignin valorization. Here, we describe P450s that catalyze the O-demethylation of lignin-derived guaiacols with different ring substitution patterns. Bacterial strains Rhodococcus rhodochrous EP4 and Rhodococcus jostii RHA1 both utilized alkylguaiacols as sole growth substrates. Transcriptomics of EP4 grown on 4-propylguaiacol (4PG) revealed the up-regulation of agcA, encoding a CYP255A1 family P450, and the aph genes, previously shown to encode a meta-cleavage pathway responsible for 4-alkylphenol catabolism. The function of the homologous pathway in RHA1 was confirmed: Deletion mutants of agcA and aphC, encoding the meta-cleavage alkylcatechol dioxygenase, grew on guaiacol but not 4PG. By contrast, deletion mutants of gcoA and pcaL, encoding a CYP255A2 family P450 and an ortho-cleavage pathway enzyme, respectively, grew on 4-propylguaiacol but not guaiacol. CYP255A1 from EP4 catalyzed the O-demethylation of 4-alkylguaiacols to 4-alkylcatechols with the following apparent specificities (kcat/KM): propyl > ethyl > methyl > guaiacol. This order largely reflected AgcA's binding affinities for the different guaiacols and was the inverse of GcoAEP4's specificities. The biocatalytic potential of AgcA was demonstrated by the ability of EP4 to grow on lignin-derived products obtained from the reductive catalytic fractionation of corn stover, depleting alkylguaiacols and alkylphenols. By identifying related P450s with complementary specificities for lignin-relevant guaiacols, this study facilitates the design of these enzymes for biocatalytic applications. We further demonstrated that the metabolic fate of the guaiacol depends on its substitution pattern, a finding that has significant implications for engineering biocatalysts to valorize lignin.

Coral-reef fishes can become more risk-averse at their poleward range limits.

As climate warms, tropical species are expanding their distribution to temperate ecosystems where they are confronted with novel predators and habitats. Predation strongly regulates ecological communities, and range-extending species that adopt an effective antipredator strategy have a higher likelihood to persist in non-native environments. Here, we test this hypothesis by comparing various proxies of antipredator and other fitness-related behaviours between range-extending tropical fishes and native-temperate fishes at multiple sites across a 730 km latitudinal range. Although some behavioural proxies of risk aversion remained unaltered for individual tropical fish species, in general they became more risk-averse (increased sheltering and/or flight initiation distance), and their activity level decreased poleward. Nevertheless, they did not experience a decline in body condition or feeding rate in their temperate ranges. Temperate fishes did not show a consistently altered pattern in their behaviours across range locations, even though one species increased its flight initiation distance at the warm-temperate location and another one had lowest activity levels at the coldest range location. The maintenance of feeding and bite rate combined with a decreased activity level and increased sheltering may be behavioural strategies adopted by range-extending tropical fishes, to preserve energy and maintain fitness in their novel temperate ecosystems.

Ocean warming and acidification degrade shoaling performance and lateralization of novel tropical-temperate fish shoals.

Gregarious behaviours are common in animals and provide various benefits such as food acquisition and protection against predators. Many gregarious tropical species are shifting poleward under current ocean warming, creating novel species and social interactions with local temperate taxa. However, how the dynamics of these novel shoals might be altered by future ocean warming and acidification remains untested. Here we evaluate how novel species interactions, ocean acidification and warming affect shoaling dynamics, motor lateralization and boldness of range-extending tropical and co-shoaling temperate fishes under controlled laboratory conditions. Fishes were exposed to 1 of 12 treatments (combinations of three temperature levels, two pCO2  levels and two shoal type levels: mixed species or temperate only) for 38 days. Lateralization (a measure of asymmetric expression of cognitive function in group coordination and predator escape) of tropical and temperate species was right-side biased under present-day conditions, but side bias significantly diminished in tropical and temperate fishes under ocean acidification. Ocean acidification also decreased shoal cohesion irrespective of shoaling type, with mixed-species shoals showing significantly lower cohesion than temperate-only shoals irrespective of climate stressors. Tropical fish became bolder under ocean acidification (after 4 weeks), and temperate fish became bolder with increasing temperature, while ocean acidification dampened temperate fish boldness. Our findings highlight the direct effect of climate stressors on fish behaviour and the interplay with the indirect effects of novel species interactions. Because strong shoal cohesion and lateralization are key determinants of species fitness, their degradation under ocean warming and acidification could adversely affect species performance in novel assemblages in a future ocean, and might slow down tropical species range extensions.

Behavioural generalism could facilitate coexistence of tropical and temperate fishes under climate change.

Coral-reef fishes are shifting their distributions poleward in response to human-mediated ocean warming; yet, the consequences for recipient temperate fish communities remain poorly understood. Behavioural modification is often the first response of species to environmental change, but we know little about how this might shape the ongoing colonisation by tropical fishes of temperate-latitude ecosystems under climate change. In a global hotspot of ocean warming (southeast Australia), we quantified 14 behavioural traits of invading tropical and local co-occurring temperate fishes at 10 sites across a 730 km latitudinal gradient as a proxy of species behavioural niche space in different climate ranges (subtropical, warm-temperate and cold-temperate). We found that tropical fishes (four species) modified their behavioural niches as well as increased their overall behavioural niche breadth in their novel temperate ranges where temperate species predominate, but maintained a moderate to high niche segregation with native temperate species across latitudinal range position. Temperate species (three co-occurring species) also modified their niches, but in contrast to tropical species, experienced an increased niche breadth towards subtropical ranges. Alterations to feeding and shoaling behaviours contributed most to niche modifications in tropical and temperate species, while behaviours related to alertness and escape from potential threats contributed least. We here show that at warmer and colder range edges where community structures are being reshuffled due to climate change, behavioural generalism and niche modification are potential mechanisms adopted by tropical range extenders and native temperate fishes to adjust to novel species interactions under climate change.

Bright spots among the world's coral reefs.

Ongoing declines in the structure and function of the world's coral reefs require novel approaches to sustain these ecosystems and the millions of people who depend on them3. A presently unexplored approach that draws on theory and practice in human health and rural development is to systematically identify and learn from the 'outliers'­places where ecosystems are substantially better ('bright spots') or worse ('dark spots') than expected, given the environmental conditions and socioeconomic drivers they are exposed to. Here we compile data from more than 2,500 reefs worldwide and develop a Bayesian hierarchical model to generate expectations of how standing stocks of reef fish biomass are related to 18 socioeconomic drivers and environmental conditions. We identify 15 bright spots and 35 dark spots among our global survey of coral reefs, defined as sites that have biomass levels more than two standard deviations from expectations. Importantly, bright spots are not simply comprised of remote areas with low fishing pressure; they include localities where human populations and use of ecosystem resources is high, potentially providing insights into how communities have successfully confronted strong drivers of change. Conversely, dark spots are not necessarily the sites with the lowest absolute biomass and even include some remote, uninhabited locations often considered near pristine6. We surveyed local experts about social, institutional, and environmental conditions at these sites to reveal that bright spots are characterized by strong sociocultural institutions such as customary taboos and marine tenure, high levels of local engagement in management, high dependence on marine resources, and beneficial environmental conditions such as deep-water refuges. Alternatively, dark spots are characterized by intensive capture and storage technology and a recent history of environmental shocks. Our results suggest that investments in strengthening fisheries governance, particularly aspects such as participation and property rights, could facilitate innovative conservation actions that help communities defy expectations of global reef degradation.

Recruiting a long way from home: Domino damselfish Dascyllus trimaculatus can detect new temperate coral habitat and conspecifics.

Climate change is driving tropicalisation of temperate reefs, yet it is unclear how range-shifting tropical fishes locate suitable habitat. The authors tested whether juvenile tropical damsels Dascyllus trimaculatus could detect rare coral habitat (Pocillopora aliciae) and conspecifics on temperate rocky reefs using olfactory and visual preference experiments. For all cues, individuals selected and spent more time than expected in aquarium areas with the cue present, which included coral odour and conspecific odour and visual cues. This indicates that vagrant coral-reef fish can detect rare suitable habitat outside of their natal range.

Searching for seadragons: predicting micro-habitat use for the common (weedy) seadragon (Phyllopteryx taeniolatus) based on habitat and prey.

Habitat associations can be critical predictors of larger-scale organism distributions and range shifts. Here the authors consider how a critical habitat, kelp (Ecklonia radiata) and prey (mysid crustacean swarms), can influence small- and large-scale distribution on the iconic common (weedy) seadragon (Phyllopteryx taeniolatus:Syngnathidae). P. taeniolatus are charismatic fish endemic to the temperate reefs of southern Australia, reported to range from Geraldton, Western Australia (28.7667°S, 114.6167°E) around southern Australia to Port Stephens, New South Wales (32.614369°S, 152.325676°E). The authors test a previously developed model of seadragon habitat preferences to predict P. taeniolatus occurrence within four sites from Sydney to the northern limit of their range in eastern Australia. They determined that P. taeniolatus associations with Ecklonia and mysid shrimp can be extrapolated across multiple sites to predict the occurrence of individual P. taeniolatus within a location/site. For instance, the authors demonstrated a significant positive relationship between the density of mysid swarms and the density of P. taeniolatus, evident across all sites despite large differences in the density of mysid swarms among sites. The findings are the first to model P. taeniolatus habitat associations across multiple sites to the northern limit of their range and have applications in protecting P. taeniolatus populations and how they may respond under climate change scenarios, such as poleward kelp retractions.

Space use by the endemic common (weedy) seadragon (Phyllopteryx taeniolatus): influence of habitat and prey.

The weedy seadragon (Phyllopteryx taeniolatus: Syngnathidae) is an iconic fish endemic to the southern coastal waters of Australia. The authors of this study analysed the habitat preferences and factors influencing microhabitat selection by P. taeniolatus in a population from Kurnell, NSW, Australia. Using field surveys and the resource selection probability function, the authors determined that P. taeniolatus significantly preferred kelp (Ecklonia radiata)-dominated habitat and avoided rock-dominated habitat. They showed P. taeniolatus preferred habitat of between 40% and 80% coverage of Ecklonia, while avoiding areas of <20% cover. Furthermore, across all habitats, mysid prey availability significantly influenced P. taeniolatus habitat selection. The strong dependence of P. taeniolatus on Ecklonia habitat shown in this study was previously untested and highlights that reductions in Ecklonia cover under climate change, or impacts from increasing urbanisation, may render seadragon populations vulnerable to declines.

Opposing life stage-specific effects of ocean warming at source and sink populations of range-shifting coral-reef fishes.

Climate change is altering the latitudinal distributions of species, with their capacity to keep pace with a shifting climate depending on the stochastic expression of population growth rates, and the influence of compensatory density feedback on age-specific survival rates. We use population-abundance time series at the leading edge of an expanding species' range to quantify the contribution of stochastic environmental drivers and density feedbacks to the dynamics of life stage-specific population growth. Using a tropical, range-shifting Indo-Pacific damselfish (Abudefduf vaigiensis) as a model organism, we applied variants of the phenomenological Gompertz-logistic model to a 14-year dataset to quantify the relative importance of density feedback and stochastic environmental drivers on the separate and aggregated population growth rates of settler and juvenile life stages. The top-ranked models indicated that density feedback negatively affected the growth of tropical settlers and juveniles. Rates of settlement were negatively linked to temperatures experienced by parents at potential source populations in the tropics, but their subsequent survival and that of juveniles increased with the temperatures experienced at the temperate sink. Including these stochastic effects doubled the deviance explained by the models, corroborating an important role of temperature. By incorporating sea-surface temperature projections for the remainder of this century into these models, we anticipate improved conditions for the population growth of juvenile coral-reef fishes, but not for settlers in temperate ecosystems. Previous research has highlighted the association between temperature and the redistribution of species. Our analyses reveal the contrasting roles of different life stages in the dynamics of range-shifting species responding to climate change, as they transition from vagrancy to residency in their novel ranges.

Gravity of human impacts mediates coral reef conservation gains.

Coral reefs provide ecosystem goods and services for millions of people in the tropics, but reef conditions are declining worldwide. Effective solutions to the crisis facing coral reefs depend in part on understanding the context under which different types of conservation benefits can be maximized. Our global analysis of nearly 1,800 tropical reefs reveals how the intensity of human impacts in the surrounding seascape, measured as a function of human population size and accessibility to reefs ("gravity"), diminishes the effectiveness of marine reserves at sustaining reef fish biomass and the presence of top predators, even where compliance with reserve rules is high. Critically, fish biomass in high-compliance marine reserves located where human impacts were intensive tended to be less than a quarter that of reserves where human impacts were low. Similarly, the probability of encountering top predators on reefs with high human impacts was close to zero, even in high-compliance marine reserves. However, we find that the relative difference between openly fished sites and reserves (what we refer to as conservation gains) are highest for fish biomass (excluding predators) where human impacts are moderate and for top predators where human impacts are low. Our results illustrate critical ecological trade-offs in meeting key conservation objectives: reserves placed where there are moderate-to-high human impacts can provide substantial conservation gains for fish biomass, yet they are unlikely to support key ecosystem functions like higher-order predation, which is more prevalent in reserve locations with low human impacts.

Trophic niche segregation allows range-extending coral reef fishes to co-exist with temperate species under climate change.

Changing climate is forcing many terrestrial and marine species to extend their ranges poleward to stay within the bounds of their thermal tolerances. However, when such species enter higher latitude ecosystems, they engage in novel interactions with local species, such as altered predator-prey dynamics and competition for food. Here, we evaluate the trophic overlap between range-extending and local fish species along the east coast of temperate Australia, a hotspot for ocean warming and species range extensions. Stable isotope ratios (δ15 N and δ13 C) of muscle tissue and stomach content analysis were used to quantify overlap of trophic niche space between vagrant tropical and local temperate fish communities along a 730 km (6°) latitudinal gradient. Our study shows that in recipient temperate ecosystems, sympatric tropical and temperate species do not overlap significantly in their diet-even though they forage on broadly similar prey groups-and are therefore unlikely to compete for trophic niche space. The tropical and temperate species we studied, which are commonly found in shallow-water coastal environments, exhibited moderately broad niche breadths and local-scale dietary plasticity, indicating trophic generalism. We posit that because these species are generalists, they can co-exist under current climate change, facilitating the existence of novel community structures.

Dietary generalism accelerates arrival and persistence of coral-reef fishes in their novel ranges under climate change.

Climate change is redistributing marine and terrestrial species globally. Life-history traits mediate the ability of species to cope with novel environmental conditions, and can be used to gauge the potential redistribution of taxa facing the challenges of a changing climate. However, it is unclear whether the same traits are important across different stages of range shifts (arrival, population increase, persistence). To test which life-history traits most mediate the process of range extension, we used a 16-year dataset of 35 range-extending coral-reef fish species and quantified the importance of various traits on the arrival time (earliness) and degree of persistence (prevalence and patchiness) at higher latitudes. We show that traits predisposing species to shift their range more rapidly (large body size, broad latitudinal range, long dispersal duration) did not drive the early stages of redistribution. Instead, we found that as diet breadth increased, the initial arrival and establishment (prevalence and patchiness) of climate migrant species in temperate locations occurred earlier. While the initial incursion of range-shifting species depends on traits associated with dispersal potential, subsequent establishment hinges more on a species' ability to exploit novel food resources locally. These results highlight that generalist species that can best adapt to novel food sources might be most successful in a future ocean.

Predation avoidance and foraging efficiency contribute to mixed-species shoaling by tropical and temperate fishes.

The formation of mixed-species social groups, whereby heterospecifics form and maintain either transient or stable groups with each other, can confer substantial fitness benefits to individuals. Such benefits may arise via multiple mechanisms associated with both predation avoidance and foraging efficiency. In fishes, mixed-species shoaling reportedly occurs where displaced tropical species (known as "vagrants") interact with resident temperate species, although little is known about the nature and frequency of such interactions. To investigate this phenomenon, we used displaced tropical Indo-pacific Sergeant Abudefduf vaigiensis settling in temperate south-eastern Australia as a model system. Underwater visual surveys revealed shoal composition and size differed significantly between open-water and reef habitats, with shoals in open habitats being larger and more speciose. Shoals containing A. vaigiensis were mainly mixed-species, and larger and more speciose in open habitats than nearer to reef. Since both foraging efficiency (via access to plankton) and predation threat likely increase with increasing distance from reef habitat, we suggest that mixed-species shoaling mitigates predation risk whilst allowing increased foraging opportunities for A. vaigiensis in open areas. These findings provide support for the importance of mixed-species shoaling to the persistence of tropical reef fishes in temperate regions.

Community-wide scan identifies fish species associated with coral reef services across the Indo-Pacific.

Determining whether many functionally complementary species or only a subset of key species are necessary to maintain ecosystem functioning and services is a critical question in community ecology and biodiversity conservation. Identifying such key species remains challenging, especially in the tropics where many species co-occur and can potentially support the same or different processes. Here, we developed a new community-wide scan (CWS) approach, analogous to the genome-wide scan, to identify fish species that significantly contribute, beyond the socio-environmental and species richness effects, to the biomass and coral cover on Indo-Pacific reefs. We found that only a limited set of species (51 out of approx. 400, approx. 13%), belonging to various functional groups and evolutionary lineages, are strongly and positively associated with fish biomass and live coral cover. Many of these species have not previously been identified as functionally important, and thus may be involved in unknown, yet important, biological mechanisms that help sustain healthy and productive coral reefs. CWS has the potential to reveal species that are key to ecosystem functioning and services and to guide management strategies as well as new experiments to decipher underlying causal ecological processes.

'Stick with your own kind, or hang with the locals?' Implications of shoaling strategy for tropical reef fish on a range-expansion frontline.

Range shifts of tropical marine species to temperate latitudes are predicted to increase as a consequence of climate change. To date, the research focus on climate-mediated range shifts has been predominately dealt with the physiological capacity of tropical species to cope with the thermal challenges imposed by temperate latitudes. Behavioural traits of individuals in the novel temperate environment have not previously been investigated, however, they are also likely to play a key role in determining the establishment success of individual species at the range-expansion forefront. The aim of this study was to investigate the effect of shoaling strategy on the performance of juvenile tropical reef fishes that recruit annually to temperate waters off the south east coast of Australia. Specifically, we compared body-size distributions and the seasonal decline in abundance through time of juvenile tropical fishes that shoaled with native temperate species ('mixed' shoals) to those that shoaled only with conspecifics (as would be the case in their tropical range). We found that shoaling with temperate native species benefitted juvenile tropical reef fishes, with individuals in 'mixed' shoals attaining larger body-sizes over the season than those in 'tropical-only' shoals. This benefit in terms of population body-size distributions was accompanied by greater social cohesion of 'mixed' shoals across the season. Our results highlight the impact that sociality and behavioural plasticity are likely to play in determining the impact on native fish communities of climate-induced range expansion of coral reef fishes.

Temperature influences habitat preference of coral reef fishes: Will generalists become more specialised in a warming ocean?

Climate change is expected to pose a significant risk to species that exhibit strong behavioural preferences for specific habitat types, with generalist species assumed to be less vulnerable. In this study, we conducted habitat choice experiments to determine how water temperature influences habitat preference for three common species of coral reef damselfish (Pomacentridae) that differ in their levels of habitat specialisation. The lemon damselfish Pomacentrus moluccensis, a habitat specialist, consistently selected complex coral habitat across all temperature treatments (selected based on local average seasonal temperatures naturally experienced in situ: ambient winter 22°C; ambient summer 28°C; and elevated 31°C). Unexpectedly, the neon damselfish Pomacentrus coelestis and scissortail sergeant Abudefduf sexfasciatus, both of which have more generalist habitat associations, developed strong habitat preferences (for complex coral and boulder habitat, respectively) at the elevated temperature treatment (31°C) compared to no single preferred habitat at 22°C or 28°C. The observed shifts in habitat preference with temperature suggest that we may be currently underestimating the vulnerability of some habitat generalists to climate change and highlight that the ongoing loss of complex live coral through coral bleaching could further exacerbate resource overlap and species competition in ways not currently considered in climate change models.