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Webb, E. A.

Publications and source records attributed to Webb, E. A..

9 recordsLinked to original sources

Spatiotemporal conservation of bacteria in the Trichodesmium microbiome

Trichodesmium is a major nitrogen fixer in the nutrient-limited tropical and subtropical ocean, contributing up to 50% of new nitrogen and driving primary productivity. Trichodesmium filaments often aggregate into colonies that host a diverse consortium of microbes that can influence their activity and physiology; however, it is unclear what factors shape community structure. To address this, we used metagenomics of Trichodesmium colonies collected across the subtropical and tropical Atlantic Ocean to examine how Trichodesmium diversity and abundance shape epibiont community composition. Heterotrophic metagenome-assembled-genomes from Trichodesmium colonies and genes specific to the major Trichodesmium subclades were read mapped against Trichodesmium metagenomes from the North Atlantic, the Western Tropical South Pacific, and the Red Sea. Using this global survey of Trichodesmium subclade distribution patterns and their enriched heterotrophs, we found unique core microbiomes that are specific to Trichodesmium subclades across ocean basins. Genome characteristics of the enriched heterotrophs reveal a genomically small, slow-growing community consistently associates with colonies. Our results suggest that Trichodesmium genotype may influence epibiont community structure, indicating a role for deterministic processes in shaping the microbiome.

microbiology↗

Genomics and biogeography of novel Trichodesmium isolates from the Sargasso Sea

The cyanobacterium Trichodesmium is important in ocean nitrogen (N) and carbon (C) biogeochemistry through fixation of N2 gas and CO2 into NH3, amino acids, and carbohydrates. These essential compounds then fuel primary productivity, bacterial and grazing communities, and upper trophic levels. In the oligotrophic surface oceans, they are one of few taxa capable of utilizing both atmospheric C and N. Trichodesmium fall into 2 diazotrophic phylogenomic clades named "Thieb" and "Tery" after the most well characterized botanical species, Trichodesmium thiebautii and Trichodesmium erythraeum. Thieb is commonly the most abundant clade in all major ocean regions and paradoxically, is also the least characterized in terms of morphology, genomics, and ecophysiology due to underrepresentation in culture collections. In this study, 25 novel strains of Thieb were enriched in culture from the Sargasso Sea, phenotypically characterized, and whole genome sequenced. Cultures were found to belong to four well-supported subclades (i.e., ThiebA, ThiebB, ThiebC, and ThiebD). Trichodesmium subclades were found to have environmentally relevant differences in colony shape, gene content, cell dimensions, and inferred temperature range. Specifically, ThiebD and ThiebC had the largest biovolumes whereas ThiebA and ThiebB had the smallest. Importantly ThiebB also became relatively more abundant only in bulk seawater metagenomes collected at high temperature (>28{degrees}C). This finding indicates that as ocean warming continues, there may be shifts in Trichodesmium community structure towards smaller cells, which would affect overall C+N standing stocks.

microbiology↗

Global distribution of cultivated Trichodesmium subclades revealed by multi-omics

Trichodesmium is an important oceanic N2 fixing cyanobacterial genus that has been shown to provide up to 50% of new N to oligotrophic regimes. Despite its importance, we know relatively little about the genomic potential and functional diversity of the two major clades of Trichodesmium (Clade I and Clade III, hereafter Thieb and Tery, respectively). With the expanded collection of Trichodesmium isolates in the USCTCC (University of Southern California Trichodesmium Culture Collection), we sequenced genomes from seven cultivated strains to further characterize the genomic diversity within the genus. For example, sequencing the genome of the "gigantic", red Trichodesmium contortum surprisingly shows that they are closely related to the smallest Trichodesmium clade, TeryA. The high genomic identity between TeryA and T. contortum (>98.5% ANI) and lack of implicated auxiliary genes suggests their large biovolume differences might be transcriptional or epigenetic in origin. Furthermore, these data show that using Tery-subclades are a more accurate designation than the classical Trichodesmium species delineation. Finally, we placed the analysis of these genomes in an ecological context via read mapping with globally distributed RNA and DNA datasets. Our data show that Tery clades (A&B) are both lower in relative abundance compared to Thieb in global oceans, generally co-occur when detected in the field, and are highly linked to decreased salinity and increased temperatures, especially for sampling locations in the Bay of Bengal. Lastly, even though TeryA members are undersaturated with respect to current CO2 concentrations, our phenotypic and biogeography data suggests that salinity/ocean color could limit their predicted global impact as climate changes.

microbiology↗

Spatial and Temporal Resolution of Cyanobacterial Bloom Chemistry Reveals an Open-Ocean Trichodesmium thiebautii as a Talented Producer of Specialized Metabolites

While the ecological role that Trichodesmium sp. play in nitrogen fixation has been widely studied, little information is available on potential specialized metabolites that are associated with blooms and standing stock Trichodesmium colonies. While a collection of biological material from a T. thiebautii bloom event from North Padre Island, TX in 2014 indicated that this species was a prolific producer of chlorinated specialized metabolites, additional spatial and temporal resolution was needed. We have completed these metabolite comparison studies, detailed in the current report, utilizing LC-MS/MS-based molecular networking to visualize and annotate the specialized metabolite composition of these Trichodesmium blooms and colonies in the Gulf of Mexico (GoM) and other waters. Our results showed that T. thiebautii blooms and colonies found in the GoM have a remarkably consistent specialized metabolome. Additionally, we isolated and characterized one new macrocyclic compound from T. thiebautii, trichothilone A (1), which was also detected in three independent cultures of T. erythraeum. Genome mining identified genes predicted to synthesize certain functional groups in the T. thiebautii metabolites. These results provoke intriguing questions of how these specialized metabolites affect Trichodesmium ecophysiology, symbioses with marine invertebrates, and niche development in the global oligotrophic ocean. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=140 SRC="FIGDIR/small/569781v1_ufig1.gif" ALT="Figure 1"> View larger version (52K): org.highwire.dtl.DTLVardef@7bce53org.highwire.dtl.DTLVardef@26d052org.highwire.dtl.DTLVardef@36d44aorg.highwire.dtl.DTLVardef@1ab85fb_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

A novel, N2-fixing cyanobacterium present and active in the global oceans

Marine N2-fixing cyanobacteria, including the unicellular genus Crocosphaera, are considered keystone species in marine food webs. Crocosphaera are globally distributed and provide new sources of nitrogen (N) and carbon (C), which fuel oligotrophic microbial communities and upper trophic levels. Despite their ecosystem importance, only one species, Crocosphaera watsonii, has ever been identified and characterized as widespread in the oligotrophic oceans. Herein, we present a novel species, candidatus Crocosphaera waterburyi (C. waterburyi hereafter), enriched from the North Pacific Ocean, active in situ, and globally distributed in environmental datasets. C. waterburyi is morphologically, phylogenetically, and physiologically distinct from C. watsonii; therefore, description of this novel species provides a new window into previously uncharacterized diversity and ecology of unicellular N2-fixing cyanobacterial taxa and further highlights their importance in the global N cycle.

genomics↗

Importance of Mobile Genetic Element Immunity in Numerically Abundant Trichodesmium Clades

The colony-forming cyanobacteria Trichodesmium spp. are considered one of the most important nitrogen-fixing genera in the warm, low nutrient, open ocean. Despite this central biogeochemical role, many questions about their evolution, physiology, and trophic interactions remain unanswered. To address these questions, we describe the genetic potential of the genus via significantly improved genomic assemblies of strains Trichodesmium thiebautii H94, Trichodesmium erythraeum 2175, and 17 new Trichodesmium metagenome-assembled genomes (MAGs, >50% complete) from hand-picked, Trichodesmium colonies spanning the Atlantic Ocean. Phylogenomics identified [~]four N2 fixing clades of Trichodesmium across the transect, with T. thiebautii dominating the colony-specific reads. Pangenomic analyses showed that all T. thiebautii MAGs are enriched in defense mechanisms and encode a vertically inherited Type III-B Clustered Regularly Interspaced Short Palindromic Repeats and associated protein-based immunity system (CRISPR-Cas hereafter). Surprisingly, this CRISPR-Cas system was absent in all T. erythraeum genomes and MAGs, vertically inherited by T. thiebautii, and correlated with increased signatures of horizontal gene transfer. Multiple lines of evidence indicate that the CRISPR-Cas system is functional in situ: 1. Trichodesmium CRISPR spacer sequences with 100% identical hits to field-assembled, putative phage genome fragments were identified, 2. High Trichodesmium spacer sequence variability indicating rapid adaptation, and 3. metaproteomic and transcriptomic expression analyses detecting the CRISPR-Cas system components in Trichodesmium colonies from the Atlantic and Pacific Oceans. These data suggest that phage or mobile genetic element immunity in T. thiebautii could contribute to their success, gene diversity, and numerical dominance over T. erythraeum in the oceans, thus warranting further Trichodesmium virome investigations. Significance statementOur work identifies CRISPR-Cas immunity as a phylogenetically distinct, environmentally expressed factor in the speciation of closely related N2-fixing Trichodesmium clades. These findings suggest that differential phage predation and resistance could be a previously overlooked selective pressure in the genus, potentially leading to the current numerical dominance of T. thiebautii over T. erythraeum in the oceans. Furthermore, while the currently CO2-limited T. erythraeum is expected to be a winner of anthropogenic climate change, their predicted higher phage sensitivity than T. thiebautii could challenge this outcome.

microbiology↗

Alphaproteobacteria facilitate Trichodesmium community trimethylamine utilization

In the surface waters of the warm oligotrophic ocean, filaments and aggregated colonies of the nitrogen (N)-fixing cyanobacterium Trichodesmium create microscale nutrient-rich oases. These hotspots fuel primary productivity and harbor a diverse consortium of heterotrophs. Interactions with associated microbiota can affect the physiology of Trichodesmium, often in ways that have been predicted to support its growth. Recently, it was found that trimethylamine (TMA), a globally-abundant organic N compound, inhibits N2 fixation in cultures of Trichodesmium without impairing growth rate, suggesting that Trichodesmium receives nitrogen from TMA. In this study, 15N-TMA DNA stable isotope probing (SIP) of a Trichodesmium enrichment was employed to further investigate TMA metabolism and determine if TMA-N is incorporated directly or secondarily via cross-feeding facilitated by microbial associates. Herein we identify two members of the marine Roseobacter clade (MRC) of Alphaproteobacteria as the likely metabolizers of TMA and provide genomic evidence that they converted TMA into a more readily available form of N, e.g., NH4+, which was subsequently used by Trichodesmium and the rest of the community. The results implicate microbiome-mediated carbon (C) and N transformations in modulating N2 fixation, and thus highlight the involvement of host-associated heterotrophs in global biogeochemical cycling.

microbiology↗

Mechanisms and heterogeneity of mineral use by natural colonies of the cyanobacterium Trichodesmium

The keystone marine nitrogen fixer Trichodesmium thrives in high dust environments, and while experimental observations suggest that Trichodesmium colonies can access the essential nutrient iron from dust particles, it is not known the extent to which this occurs in the field. Here we demonstrate that Trichodesmium colonies actively process mineral particles in nature with direct molecular impacts. Microscopy and synchrotron-based imaging demonstrated heterogeneous associations with particles consistent with iron oxide and iron silicate minerals. Metaproteomic analysis of individual colonies revealed enrichment of biogeochemically-relevant proteins including photosynthesis proteins and metalloproteins containing iron, nickel, copper and zinc when particles were present. The iron-storage protein ferritin was particularly enriched implying accumulation of particle-derived iron, and multiple iron acquisition pathways including Fe(II), Fe(III), and Fe-siderophore transporters were engaged, including evidence of superoxide-driven particle dissolution. While the particles clearly provided iron, there was also evidence that the concentrated metals had toxic effects. The molecular mechanisms allowing Trichodesmium to interact with particulate minerals are fundamental to its success and global impact on nitrogen biogeochemistry, and may contribute to the leaching of particulate trace metals with implications for global iron and carbon cycling.

ecology↗

Dual thermal ecotypes detected within a nearly genetically-identical population of the unicellular marine cyanobacterium Synechococcus

The extent and ecological significance of intraspecific diversity within marine microbial populations is still poorly understood, and it remains unclear if such strain-level microdiversity will affect fitness and persistence in a rapidly changing ocean environment. In this study, we cultured 11 sympatric strains of the ubiquitous marine picocyanobacterium Synechococcus isolated from a Narragansett Bay (Rhode Island, USA) phytoplankton community thermal selection experiment. Despite all 11 isolates being highly similar (with average nucleotide identities of >99.9%, with 98.6-100% of the genome aligning), thermal performance curves revealed selection at warm and cool temperatures had subdivided the initial population into thermotypes with pronounced differences in maximum growth temperatures. Within the fine-scale genetic diversity that did exist within this population, the two divergent thermal ecotypes differed at a locus containing genes for the phycobilisome antenna complex. Our study demonstrates that present-day marine microbial populations can contain microdiversity in the form of cryptic but environmentally-relevant thermotypes that may increase their resilience to future rising temperatures. SignificanceNumerous studies exist comparing the responses of distinct taxonomic groups of marine microbes to a warming ocean (interspecific thermal diversity). For example, Synechococcus, a nearly globally distributed unicellular marine picocyanobacterium that makes significant contributions to oceanic primary productivity, contains numerous taxonomically distinct lineages with well documented temperature relationships. Little is known though about the diversity of functional responses to temperature within a given population where genetic similarity is high (intraspecific thermal diversity). This study suggests that understanding the extent of this functional intraspecific microdiversity is an essential prerequisite to predicting the resilience of biogeochemically essential microbial groups such as marine Synechococcus to a changing climate.

microbiology↗