Russell Y. Neches

Functional biogeography of ocean microbes revealed through non-negative matrix factorization.

The direct “metagenomic” sequencing of genomic material from complex assemblages of bacteria, archaea, viruses and microeukaryotes has yielded new insights into the structure of microbial communities. For example, analysis of metagenomic data has revealed the existence of previously unknown microbial taxa whose spatial distributions are limited by environmental conditions, ecological competition, and dispersal mechanisms. However, differences in genotypes that might lead biologists to designate two microbes as taxonomically distinct need not necessarily imply differences in ecological function. Hence, there is a growing need for large-scale analysis of the distribution of microbial function across habitats. Here, we present a framework for investigating the biogeography of microbial function by analyzing the distribution of protein families inferred from environmental sequence data across a global collection of sites. We map over 6,000,000 protein sequences from unassembled reads from the Global Ocean Survey dataset to protein families, generating a protein family relative abundance matrix that describes the distribution of each protein family across sites. We then use non-negative matrix factorization (NMF) to approximate these protein family profiles as linear combinations of a small number of ecological components. Each component has a characteristic functional profile and site profile. Our approach identifies common functional signatures within several of the components. We use our method as a filter to estimate functional distance between sites, and find that an NMF-filtered measure of functional distance is more strongly correlated with environmental distance than a comparable PCA-filtered measure. We also find that functional distance is more strongly correlated with environmental distance than with geographic distance, in agreement with prior studies. We identify similar protein functions in several components and suggest that functional co-occurrence across metagenomic samples could lead to future methods for de-novo functional prediction. We conclude by discussing how NMF, and other dimension reduction methods, can help enable a macroscopic functional description of marine ecosystems.

A workflow for genome-wide mapping of archaeal transcription factors with ChIP-seq

Deciphering the structure of gene regulatory networks across the tree of life remains one of the major challenges in postgenomic biology. We present a novel ChIP-seq workflow for the archaea using the model organism Halobacterium salinarum sp. NRC-1 and demonstrate its application for mapping the genome-wide binding sites of natively expressed transcription factors. This end-to-end pipeline is the first protocol for ChIP-seq in archaea, with methods and tools for each stage from gene tagging to data analysis and biological discovery. Genome-wide binding sites for transcription factors with many binding sites (TfbD) are identified with sensitivity, while retaining specificity in the identification the smaller regulons (bacteriorhodopsin-activator protein). Chromosomal tagging of target proteins with a compact epitope facilitates a standardized and cost-effective workflow that is compatible with high-throughput immunoprecipitation of natively expressed transcription factors. The Pique package, an open-source bioinformatics method, is presented for identification of binding events. Relative to ChIP-Chip and qPCR, this workflow offers a robust catalog of protein–DNA binding events with improved spatial resolution and significantly decreased cost. While this study focuses on the application of ChIP-seq in H. salinarum sp. NRC-1, our workflow can also be adapted for use in other archaea and bacteria with basic genetic tools.

A pharyngeal jaw evolutionary innovation facilitated extinction in Lake Victoria cichlids

The downside of innovation Evolutionary innovation allows a species to invade a new niche or environment. Generally, the emergence of adaptive traits is thought to lead to diversification. Support for this process in nature, however, is mixed. McGee et al. show that the evolution of secondary jaws in fish may be an example of how innovation can reduce diversification (see the Perspective by Vermeij). Fish with secondary jaws are less able to rapidly ingest fish prey, which puts them at a competitive disadvantage to regularly jawed fish. Such competition could lead to reduced variation and the extinction of lineages with the trait. Science, this issue p. 1077; see also p. 1038 Jaw specialization in fish may have paradoxically reduced speciation in the African Great Lakes. [Also see Perspective by Vermeij] Evolutionary innovations, traits that give species access to previously unoccupied niches, may promote speciation and adaptive radiation. Here, we show that such innovations can also result in competitive inferiority and extinction. We present evidence that the modified pharyngeal jaws of cichlid fishes and several marine fish lineages, a classic example of evolutionary innovation, are not universally beneficial. A large-scale analysis of dietary evolution across marine fish lineages reveals that the innovation compromises access to energy-rich predator niches. We show that this competitive inferiority shaped the adaptive radiation of cichlids in Lake Tanganyika and played a pivotal and previously unrecognized role in the mass extinction of cichlid fishes in Lake Victoria after Nile perch invasion.

Evaluating genomic divergence and parallelism in replicate ecomorphs from young and old cichlid adaptive radiations

Comparative genomic studies of closely related species typically focus on single species pairs at one given stage of divergence. That makes it difficult to infer the continuum of evolutionary process during speciation and beyond. Here, we use whole‐genome resequencing to examine genomic patterns of divergence in three sympatric cichlid species pairs with very similar functional and ecological differentiation, but different ages. We find a strong signature of increasing genomic divergence with time in both the mitochondrial genome and the nuclear genome. In contrast to many other systems, we find that in these cichlids, regions of elevated relative differentiation also exhibit increased absolute differentiation. We detect a signature of convergent evolution in a comparison of outlier regions across all three species pair comparisons, but the extent of it is modest, and regions that are strongly divergent in any one pair tend to be only slightly elevated in the other pairs, consistent with a repeatable but polygenic basis of traits that characterize the ecomorphs. Our results suggest that strong functional phenotypic differentiation, as seen in all three species pairs, is generally associated with a clear signature of genomic divergence, even in the youngest species pair.

The Common Oceanographer: Crowdsourcing the Collection of Oceanographic Data

We live on a vast, underexplored planet that is largely ocean. Despite modern technology, Global Positioning System (GPS) navigation, and advanced engineering of ocean vessels, the ocean is unforgiving, especially in rough weather. Coastal ocean navigation, with risks of running aground and inconsistent weather and sea patterns, can also be challenging and hazardous. In 2012, more than 100 international incidents of ships sinking, foundering, grounding, or being lost at sea were reported (http://en.wikipedia.org/wiki/List_of_shipwrecks_in_2012). Even a modern jetliner can disappear in the ocean with little or no trace [1], and the current costs and uncertainty associated with search and rescue make the prospects of finding an object in the middle of the ocean daunting [2]. Notwithstanding satellite constellations, autonomous vehicles, and more than 300 research vessels worldwide (www.wikipedia.org/wiki/List_of_research_vessels_by_country), we lack fundamental data relating to our oceans. These missing data hamper our ability to make basic predictions about ocean weather, narrow the trajectories of floating objects, or estimate the impact of ocean acidification and other physical, biological, and chemical characteristics of the worlds oceans. To cope with this problem, scientists make probabilistic inferences by synthesizing models with incomplete data. Probabilistic modeling works well for certain questions of interest to the scientific community, but it is difficult to extract unambiguous policy recommendations from this approach. The models can answer important questions about trends and tendencies among large numbers of events but often cannot offer much insight into specific events. For example, probabilistic models can tell us with some precision the extent to which storm activity will be intensified by global climate change but cannot yet attribute the severity of a particular storm to climate change. Probabilistic modeling can provide important insights into the global traffic patterns of floating debris but is not of much help to search-and-rescue personnel struggling to learn the likely trajectory of a particular piece of debris left by a wreck. Oceanographic data are incomplete because it is financially and logistically impractical to sample everywhere. Scientists typically sample over time, floating with the currents and observing their temporal evolution (the Langrangian approach), or they sample across space to cover a gradient of conditions—such as temperature or nutrients (the Eulerian approach). These observational paradigms have various strengths and weaknesses, but their fundamental weakness is cost. A modern ocean research vessel typically costs more than US

Spatially extensive microbial biogeography of the Indian Ocean provides insights into the unique community structure of a pristine coral atoll.

30,000 per day to operate—excluding the full cost of scientists, engineers, and the cost of the research itself. Even an aggressive expansion of oceanographic research budgets would not do much to improve the precision of our probabilistic models, let alone to quickly and more accurately locate missing objects in the huge, moving, three-dimensional seascape. Emerging autonomous technologies such as underwater gliders and in situ biological samplers (e.g., environmental sample processors) help fill gaps but are cost prohibitive to scale up. Similarly, drifters (e.g., the highly successful Argo floats program) have proven very useful for better defining currents, but unless retrieved after their operational lifetime, they become floating trash, adding to a growing problem. Long-term sampling efforts such as the continuous plankton recorder in the North Sea and North Atlantic [3] provide valuable data on decadal trends and leveraged English Channel ferries to accomplish much of the sampling. Modernizing and expanding this approach is a goal of citizen science initiatives. How do we leverage cost-effective technologies and economies of scale given shrinking federal research budgets?

Growth of 48 built environment bacterial isolates on board the International Space Station (ISS)

Microorganisms act both as drivers and indicators of perturbations in the marine environment. In an effort to establish baselines to predict the response of marine habitats to environmental change, here we report a broad survey of microbial diversity across the Indian Ocean, including the first microbial samples collected in the pristine lagoon of Salomon Islands, Chagos Archipelago. This was the first large-scale ecogenomic survey aboard a private yacht employing a ‘citizen oceanography’ approach and tools and protocols easily adapted to ocean going sailboats. Our data highlighted biogeographic patterns in microbial community composition across the Indian Ocean. Samples from within the Salomon Islands lagoon contained a community which was different even from adjacent samples despite constant water exchange, driven by the dominance of the photosynthetic cyanobacterium Synechococcus. In the lagoon, Synechococcus was also responsible for driving shifts in the metatranscriptional profiles. Enrichment of transcripts related to photosynthesis and nutrient cycling indicated bottom-up controls of community structure. However a five-fold increase in viral transcripts within the lagoon during the day, suggested a concomitant top-down control by bacteriophages. Indeed, genome recruitment against Synechococcus reference genomes suggested a role of viruses in providing the ecological filter for determining the β-diversity patterns in this system.

On the intrinsic sterility of 3D printing

Background. While significant attention has been paid to the potential risk of pathogenic microbes aboard crewed spacecraft, the non-pathogenic microbes in these habitats have received less consideration. Preliminary work has demonstrated that the interior of the International Space Station (ISS) has a microbial community resembling those of built environments on Earth. Here we report the results of sending 48 bacterial strains, collected from built environments on Earth, for a growth experiment on the ISS. This project was a component of Project MERCCURI (Microbial Ecology Research Combining Citizen and University Researchers on ISS). Results. Of the 48 strains sent to the ISS, 45 of them showed similar growth in space and on Earth using a relative growth measurement adapted for microgravity. The vast majority of species tested in this experiment have also been found in culture-independent surveys of the ISS. Only one bacterial strain showed significantly different growth in space. Bacillus safensis JPL-MERTA-8-2 grew 60% better in space than on Earth. Conclusions. The majority of bacteria tested were not affected by conditions aboard the ISS in this experiment (e.g., microgravity, cosmic radiation). Further work on Bacillus safensis could lead to interesting insights on why this strain grew so much better in space.

A microbial survey of the International Space Station (ISS)

3D printers that build objects using extruded thermoplastic are quickly becoming commonplace tools in laboratories. We demonstrate that with appropriate handling, these devices are capable of producing sterile components from a non-sterile feedstock of thermoplastic without any treatment after fabrication. The fabrication process itself results in sterilization of the material. The resulting 3D printed components are suitable for a wide variety of applications, including experiments with bacteria and cell culture.

Correction: A microbial survey of the International Space Station (ISS)

Background Modern advances in sequencing technology have enabled the census of microbial members of many natural ecosystems. Recently, attention is increasingly being paid to the microbial residents of human-made, built ecosystems, both private (homes) and public (subways, office buildings, and hospitals). Here, we report results of the characterization of the microbial ecology of a singular built environment, the International Space Station (ISS). This ISS sampling involved the collection and microbial analysis (via 16S rDNA PCR) of 15 surfaces sampled by swabs onboard the ISS. This sampling was a component of Project MERCCURI (Microbial Ecology Research Combining Citizen and University Researchers on ISS). Learning more about the microbial inhabitants of the “buildings” in which we travel through space will take on increasing importance, as plans for human exploration continue, with the possibility of colonization of other planets and moons. Results Sterile swabs were used to sample 15 surfaces onboard the ISS. The sites sampled were designed to be analogous to samples collected for (1) the Wildlife of Our Homes project and (2) a study of cell phones and shoes that were concurrently being collected for another component of Project MERCCURI. Sequencing of the 16S rDNA genes amplified from DNA extracted from each swab was used to produce a census of the microbes present on each surface sampled. We compared the microbes found on the ISS swabs to those from both homes on Earth and data from the Human Microbiome Project. Conclusions While significantly different from homes on Earth and the Human Microbiome Project samples analyzed here, the microbial community composition on the ISS was more similar to home surfaces than to the human microbiome samples. The ISS surfaces are species-rich with 1,036–4,294 operational taxonomic units (OTUs per sample). There was no discernible biogeography of microbes on the 15 ISS surfaces, although this may be a reflection of the small sample size we were able to obtain.