Project description:These data are relative concentrations of targeted metabolites measured in particulate matter collected from near the Island of Hawai’i in July 2018 during the Kilauea eruption. Six stations were sampled, with station 2 representing the most geothermally impacted station closest to the lava entry. Station 6 was the most oligotrophic station. The particulate metabolites show evidence of altered community composition and metabolism at the geothermally impacted station. This station was within a phytoplankton bloom. The bloom was stimulated by lava heating deep seawater and driving upwelling, which then provided nutrients for diatom growth. See Wilson and Hawco, et. al. 2019 (DOI: 10.1126/science.aax4767) for a complete description of sample collection, phytoplankton bloom dynamics, and chemical modifications of seawater due to the eruption. Metabolites with high concentration (relative abundance per L of seawater) in the geothermally impacted station (St 2) compared to the other stations were: cytosine, hydroxyectoine, adenine, adenosine, thymine, glutamic acid, ectoine, deoxyadenosine, UDP-glucosamine, and guanosine. After normalizing to the particulate carbon concentration at each station, guanosine, glutamic acid, hydroxyectoine, ectoine, adenosine, deoxyadenosine, and UDP-glucosamine were enriched in the geothermally impacted station relative to other stations. Ectoine was the metabolite with the largest change between St 2 and St 6, regardless of normalization to L of seawater or to moles of particulate carbon, with dramatically higher concentrations in the geothermally impacted waters. Except for glutamic and proline, particulate amino acids were generally in higher concentrations in the oligotrophic station.
Project description:Microarrays are useful tools for detecting and quantifying specific functional and phylogenetic genes in natural microbial communities. In order to track uncultivated microbial genotypes and their close relatives in an environmental context, we designed and implemented a “genome proxy” microarray that targets microbial genome fragments recovered directly from the environment. Fragments consisted of sequenced clones from large-insert genomic libraries from microbial communities in Monterey Bay, the Hawaii Ocean Time-series station ALOHA, and Antarctic coastal waters. In a prototype array, we designed probe sets to thirteen of the sequenced genome fragments and to genomic regions of the cultivated cyanobacterium Prochlorococcus MED4. Each probe set consisted of multiple 70-mers, each targeting an individual ORF, and distributed along each ~40-160kbp contiguous genomic region. The targeted organisms or clones, and close relatives, were hybridized to the array both as pure DNA mixtures and as additions of cells to a background of coastal seawater. This prototype array correctly identified the presence or absence of the target organisms and their relatives in laboratory mixes, with negligible cross-hybridization to organisms having ≤~75% genomic identity. In addition, the array correctly identified target cells added to a background of environmental DNA, with a limit of detection of ~0.1% of the community, corresponding to ~10^3 cells/ml in these samples. Signal correlated to cell concentration with an R2 of 1.0 across six orders of magnitude. In addition the array could track a related strain (at 86% genomic identity to that targeted) with a linearity of R2=0.9999 and a limit of detection of ~1% of the community. Closely related genotypes were distinguishable by differing hybridization patterns across each probe set. This array’s multiple-probe, “genome-proxy” approach and consequent ability to track both target genotypes and their close relatives is important for the array’s environmental application given the recent discoveries of considerable intra-population diversity within marine microbial communities. Keywords: target addition experiment, proof-of-concept for GPL6012