Stable isotopes as tracers of trophic interactions in marine mutualistic symbioses

Mutualistic nutritional symbioses are widespread in marine ecosystems. They involve the association of a host organism (algae, protists, or marine invertebrates) with symbiotic microorganisms, such as bacteria, cyanobacteria, or dinoflagellates. Nutritional interactions between the partners are difficult to identify in symbioses because they only occur in intact associations. Stable isotope analysis (SIA) has proven to be a useful tool to highlight original nutrient sources and to trace nutrients acquired by and exchanged between the different partners of the association. However, although SIA has been extensively applied to study different marine symbiotic associations, there is no review taking into account of the different types of symbiotic associations, how they have been studied via SIA, methodological issues common among symbiotic associations, and solutions that can be transferred from one type of association with another. The present review aims to fill such gaps in the scientific literature by summarizing the current knowledge of how isotopes have been applied to key marine symbioses to unravel nutrient exchanges between partners, and by describing the difficulties in interpreting the isotopic signal. This review also focuses on the use of compound-specific stable isotope analysis and on statistical advances to analyze stable isotope data. It also highlights the knowledge gaps that would benefit from future research.info:eu-repo/semantics/publishedVersio

Diversity and function of prevalent symbiotic marine bacteria in the genus Endozoicomonas

© The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Applied Microbiology and Biotechnology 100 (2016): 8315–8324, doi:10.1007/s00253-016-7777-0.Endozoicomonas bacteria are emerging as extremely diverse and flexible symbionts of numerous marine hosts inhabiting oceans worldwide. Their hosts range from simple invertebrate species, such as sponges and corals, to complex vertebrates, such as fish. Although widely distributed, the functional role of Endozoicomonas within their host microenvironment is not well understood. In this review, we provide a summary of the currently recognized hosts of Endozoicomonas and their global distribution. Next, the potential functional roles of Endozoicomonas, particularly in light of recent microscopic, genomic, and genetic analyses, are discussed. These analyses suggest that Endozoicomonas typically reside in aggregates within host tissues, have a free-living stage due to their large genome sizes, show signs of host and local adaptation, participate in host-associated protein and carbohydrate transport and cycling, and harbour a high degree of genomic plasticity due to the large proportion of transposable elements residing in their genomes. This review will finish with a discussion on the methodological tools currently employed to study Endozoicomonas and host interactions and review future avenues for studying complex host-microbial symbioses.This work was supported by a KAUST-WHOI Post-doctoral Partnership Award to MJN and a KAUST-WHOI Special Academic Partnership Funding Reserve Award to CRV and AA. Research in this study was further supported by baseline research funds to CRV by KAUST and NSF award OCE-1233612 to AA

Physiological performance of the cold-water coral Dendrophyllia cornigera reveals its preference for temperate environments

Cold-water corals (CWCs) are key ecosystem engineers in deep-sea benthic communities around the world. Their distribution patterns are related to several abiotic and biotic factors, of which seawater temperature is arguably one of the most important due to its role in coral physiological processes. The CWC Dendrophyllia cornigera has the particular ability to thrive in several locations in which temperatures range from 11 to 17 °C, but to be apparently absent from most CWC reefs at temperatures constantly below 11 °C. This study thus aimed to assess the thermal tolerance of this CWC species, collected in the Mediterranean Sea at 12 °C, and grown at the three relevant temperatures of 8, 12, and 16 °C. This species displayed thermal tolerance to the large range of seawater temperatures investigated, but growth, calcification, respiration, and total organic carbon (TOC) fluxes severely decreased at 8 °C compared to the in situ temperature of 12 °C. Conversely, no significant differences in calcification, respiration, and TOC fluxes were observed between corals maintained at 12 and 16 °C, suggesting that the fitness of this CWC is higher in temperate rather than cold environments. The capacity to maintain physiological functions between 12 and 16 °C allows D. cornigera to be the most abundant CWC species in deep-sea ecosystems where temperatures are too warm for other CWC species (e.g., Canary Islands). This study also shows that not all CWC species occurring in the Mediterranean Sea (at deep-water temperatures of 12-14 °C) are currently living at their upper thermal tolerance limit. © 2014 Springer-Verlag Berlin Heidelber

Nitrate in the coral symbiosis: from the regulation of its assimilation to its impact on the physiology of the holobiont

In oligotrophic reef systems, coral holobionts are remarkably efficient at assimilating nitrogen through heterotrophic feeding or the uptake of dissolved inorganic nitrogen. Symbiodiniaceae are vital partners of the symbiosis for nutrient assimilation. In addition to providing translocated photosynthates, they account for most of the uptake of dissolved inorganic nitrogen. Although NO3- is the most abundant source of nitrogen in the ocean, little is known about the mechanisms regulating its assimilation by the holobiont. Coral hosts are unable to reduce nitrate as they lack the necessary enzymes, whereas Symbiodiniaceae have been shown to express the enzyme nitrate reductase (NR). However, the evidence supporting the active reduction of nitrate by the symbiotic algae during symbiosis is scarce and equivocal. This research aimed at deciphering the pathways of NO3- assimilation in both free-living Symbiodiniaceae and in hospite symbionts while also investigating the relevance of inorganic nitrogen source in physiological responses to stress. We investigated the expression and regulation of NR both in free-living Symbiodiniaceae and in in hospite symbionts using a combined western blot and qRT-PCR approach. We showed that the expression and regulation of NR in free-living Symbiodiniaceae is a dynamic and reversible process impacted by NO3- and NH4+ concentrations. Symbionts from N-depleted corals incubated with NO3- enriched seawater showed an increase in NR synthesis over time. Interestingly, NR protein synthesis did not correlate with NR gene expression, hinting towards a potential post-transcriptional regulation of the enzyme. Additionally, we investigated the impacts of inorganic N source (NO3- vs NH4+ vs N depletion) in combination with stress on the physiology of Symbiodiniaceae (photosynthetic responses, ROS and NO production). The availability of inorganic nitrogen improved photosynthetic capacity while reducing ROS production. Moreover, preliminary experiments showed that NO3- and NH4+ had differential effects on the physiological responses of Symbiodiniaceae subjected to stress

Studies With Soft Corals – Recommendations on Sample Processing and Normalization Metrics

Soft corals (Octocorallia) often constitute the second most abundant macrobenthic group on many tropical and temperate reefs. However, the gelatinous and leather-like nature of their tissue and their variable hydroskeleton entails a number of problems for tissue homogenization and data normalization. An easy and fast protocol for tissue homogenization, as well as a normalization metric that can be used to perform inter-studies or inter-species comparisons, are thus needed. In this study, we tested whether the tissue sample state before processing (frozen vs. freeze-dried samples) and the media used for tissue homogenization (0.2 μm filtered seawater; FSW vs. Milli-Q water; DI) affect the quantitative measurements of tissue descriptors (chlorophyll, protein, and Symbiodinium concentrations) in the model species Heteroxenia fuscescens. Furthermore, the suitability of dry weight (DW) and ash-free dry weight (AFDW) as size-normalizing metric was investigated across different soft coral species. Our results reveal that freeze-drying the samples and homogenizing them in DI water exhibited several benefits, namely enhancing chlorophyll and protein concentrations up to 50%, saving processing time and providing a more accurate determination of DW and AFDW. Overall, this optimized tissue processing protocol offers a more reliable quantification of tissue descriptors and reduces the chance of underestimating these parameters in soft corals. Finally, since the contribution of sclerites to the total DW of the colony can highly differ between species, we demonstrate that AFDW is a reliable metric for normalizing soft coral data, particularly when inter-species comparisons are made

Effect of salinity on the skeletal chemistry of cultured scleractinian zooxanthellate corals: Cd/Ca ratio as a potential proxy for salinity reconstruction

The effect of salinity on the elemental and isotopic skeletal composition of modern zooxanthellate scleractinian corals (Acropora sp., Montipora verrucosa and Stylophora pistillata) was investigated in order to evaluate potential salinity proxies. Corals were cultured in the laboratory at three salinities (36, 38 and 40). The other environmental parameters were kept constant. For all species analyzed, Sr/Ca, Mg/Ca, U/Ca and Li/Ca ratios were not influenced by salinity changes. The Ba/Ca ratio also lacks a systematic relationship with salinity and exhibits high inter-generic variations, up to one order of magnitude. On the contrary, the Cd/Ca ratio decreases as a function of increasing salinity, and δ18O and δ13C also presented a significant response, but with opposite trends to salinity variations. Since Cd/Ca is usually considered as an upwelling proxy, its salinity dependence could compromise the upwelling signal, unless some corrections can be carried out. Regardless, if the dependence found in the present dataset proved to be widespread and systematic, the Cd/Ca ratio could represent a promising salinometer awaiting further investigation. This study also confirmed the reliability of the well-established temperature proxies Sr/Ca, Mg/Ca and U/Ca, as these ratios were insensitive to salinity variations. Moreover, our results showed that δ18O or δ13C can be considered as reliable temperature recorders as far as the salinity effect is removed from the parameter reconstructed (e.g., temperature). Investigating the influence of salinity on the skeletal chemistry of scleractinian corals grown under controlled environmental conditions confirmed previous results, validated isotopic corrections, and identified a promising proxy of salinity