IsoSolve: an integrative framework to improve isotopic coverage and consolidate isotopic measurements by MS and/or NMR

ABSTRACTStable-isotope labeling experiments are widely used to investigate the topology and functioning of metabolic networks. Label incorporation into metabolites can be quantified using a broad range of mass spectrometry (MS)and nuclear magnetic resonance (NMR)spectroscopy methods, but in general, no single approach can completely cover isotopic space, even for small metabolites. The number of quantifiable isotopic species could be increased, and the coverage of isotopic space improved, by integrating measurements obtained by different methods; however, this approach has remained largely unexplored because no framework able to deal with partial, heterogeneous isotopic measurements has yet been developed. Here, we present a generic computational framework based on symbolic calculus that can integrate any isotopic dataset by connecting measurements to the chemical structure of the molecules. As a test case, we apply this framework to isotopic analyses of amino acids, which are ubiquitous to life, central to many biological questions, and can be analyzed by a broad range of MS and NMR methods. We demonstrate how this integrative framework helps to i) clarify and improve the coverage of isotopic space, ii) evaluate the complementarity and redundancy of different techniques, iii) consolidate isotopic datasets, iv) design experiments, and v) guide future analytical developments. This framework, which can be applied to any labeled element, isotopic tracer, metabolite, and analytical platform, has been implemented in IsoSolve (available at https://github.com/MetaSysLISBP/IsoSolve and https://pypi.org/project/IsoSolve), an open source software that can be readily integrated into data analysis pipelines.

Ecological outcomes of hybridization vary extensively in Catostomus fishes

Hybridization has been studied extensively to learn about speciation and mechanisms of reproductive isolation, but increasingly we recognize that hybridization outcomes vary geo-graphically and can depend on the environment. At the same time, hybridization can reshape biotic interactions in an ecosystem, leading to ecological shifts where hybridization occurs. Identifying how hybrid individuals function ecologically would link evolutionary outcomes of hybridization to ecological consequences, but relatively few studies have focused on ecological traits of hybrids. We described the feeding ecology of several Catostomus fish species and their hybrids by using stable isotopes (δ13C and δ15N) as a proxy for diet and habitat use, and compared two native species, an introduced species, and three interspecific hybrid crosses. We replicated this comparison spatially, including hybrids and parental species from seven rivers in the Upper Colorado River basin where parental species co-occur and the opportunity for hybridization exists. Although relationships between native species in isotopic space varied, individual native species did not fully overlap in isotopic space in any river sampled, suggesting little overlap of resource use between these historically sympatric species. The introduced species overlapped with one or both native species in every river, suggesting similar resource use and potential for competition. Hybrids occupied intermediate, matching, or more extreme (transgressive) isotopic space relative to parental species, and were isotopically variable within and among rivers. We suggest that ecological outcomes of hybridization, like genomic outcomes of hybridization, are likely to vary across locations where a pair of species hybridizes. This variation implies that hybridization might have large unpredictable, idiosyncratic ecological effects on fish assemblages where hybrids occur. Although we found little evidence that hybrids are at a disadvantage ecologically—there were no significant declines in body condition relative to parental species—it is nevertheless possible that abiotic or biotic attributes of a river might constrain the range of interspecific hybrids that are successful, thus contributing to variation in hybridization outcomes across rivers.

Niche structure of marine sponges from temperate hard-bottom habitats within Gray's Reef National Marine Sanctuary

Many species of marine sponges on tropical reefs host abundant and diverse symbiont communities capable of varied metabolic pathways. While such communities may confer a nutritional benefit to some hosts (termed High Microbial Abundance (HMA) sponges), other sympatric species host only sparse symbiont communities (termed Low Microbial Abundance (LMA) sponges) and obtain a majority of their C and N from local sources. Sponge communities are widespread across large latitudinal gradients, however, and recent evidence suggests that these symbioses may also extend beyond the tropics. We investigated the role that symbionts play in the ecology of sponges from the temperate, hard-bottom reefs of Gray's Reef National Marine Sanctuary by calculating the niche size (as standard ellipse area (SEAc)) and assessing the relative placement of five HMA and four LMA sponge species within bivariate (δ13C and δ15N) isotopic space. Although photosymbiont abundance was low across most of these species, sponges were widespread across isotopic niche space, implying that microbial metabolism confers an ecological benefit to temperate sponges by expanding host metabolic capability. To examine how these associations vary across a latitudinal gradient, we also compared the relative placement of temperate and tropical conspecifics within isotopic space. Surprisingly, shifts in sponge δ13C and δ15N values between these regions suggest a reduced reliance on symbiont-derived nutrients in temperate sponges compared with their tropical conspecifics. Despite this, symbiotic sponges in temperate systems likely have a competitive advantage, allowing them to grow and compete for space within these habitats.