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Transcriptional shifts highlight the role of nutrients in harmful brown tide dynamics (2019)

Wurch, Louie L. ; Alexander, Harriet ; Frischkorn, Kyle R. ; [et al.]

Frontiers Media

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Publication Date: 2022-05-25

Description: © The Author(s), 2019. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Wurch, L. L., Alexander, H., Frischkorn, K. R., Haley, S. T., Gobler, C. J., & Dyhrman, S. T. Transcriptional shifts highlight the role of nutrients in harmful brown tide dynamics. Frontiers in Microbiology, 10, (2019):136, doi:10.3389/fmicb.2019.00136.

Description: Harmful algal blooms (HABs) threaten ecosystems and human health worldwide. Controlling nitrogen inputs to coastal waters is a common HAB management strategy, as nutrient concentrations often suggest coastal blooms are nitrogen-limited. However, defining best nutrient management practices is a long-standing challenge: in part, because of difficulties in directly tracking the nutritional physiology of harmful species in mixed communities. Using metatranscriptome sequencing and incubation experiments, we addressed this challenge by assaying the in situ physiological ecology of the ecosystem destructive alga, Aureococcus anophagefferens. Here we show that gene markers of phosphorus deficiency were expressed in situ, and modulated by the enrichment of phosphorus, which was consistent with the observed growth rate responses. These data demonstrate the importance of phosphorus in controlling brown-tide dynamics, suggesting that phosphorus, in addition to nitrogen, should be evaluated in the management and mitigation of these blooms. Given that nutrient concentrations alone were suggestive of a nitrogen-limited ecosystem, this study underscores the value of directly assaying harmful algae in situ for the development of management strategies.

Description: This research was funded by NOAA Grant NA15NOS4780199 (SD), NA09NOA4780206 (SD and CG), and NA15NOS4780183 (CG) through the ECOHAB Program, publication number ECO929. Partial support was also provided by the World Surf League through the Columbia Center for Climate and Life, the Woods Hole Oceanographic Institution Coastal Ocean Institute, and the Link Foundation. Kyle Frischkorn was funded under a National Science Foundation Graduate Research Fellowship.

Keywords: harmful algal bloom ; Aureococcus anophagefferens ; brown tide ; nutrient physiology ; metatranscriptomics

Repository Name: Woods Hole Open Access Server

Type: Article

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Identifying reference genes with stable expression from high throughput sequence data (2014)

Alexander, Harriet ; Jenkins, Bethany D. ; Rynearson, Tatiana A. ; [et al.]

Frontiers Media

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Publication Date: 2022-05-26

Description: © The Author(s), 2012. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Frontiers in Microbiology 3 (2012): 385, doi:10.3389/fmicb.2012.00385.

Description: Genes that are constitutively expressed across multiple environmental stimuli are crucial to quantifying differentially expressed genes, particularly when employing quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) assays. However, the identification of these potential reference genes in non-model organisms is challenging and is often guided by expression patterns in distantly related organisms. Here, transcriptome datasets from the diatom Thalassiosira pseudonana grown under replete, phosphorus-limited, iron-limited, and phosphorus and iron co-limited nutrient regimes were analyzed through literature-based searches for homologous reference genes, k-means clustering, and analysis of sequence counts (ASC) to identify putative reference genes. A total of 9759 genes were identified and screened for stable expression. Literature-based searches surveyed 18 generally accepted reference genes, revealing 101 homologs in T. pseudonana with variable expression and a wide range of mean tags per million. k-means analysis parsed the whole transcriptome into 15 clusters. The two most stable clusters contained 709 genes, but still had distinct patterns in expression. ASC analyses identified 179 genes that were stably expressed (posterior probability 〈 0.1 for 1.25 fold change). Genes known to have a stable expression pattern across the test treatments, like actin, were identified in this pool of 179 candidate genes. ASC can be employed on data without biological replicates and was more robust than the k-means approach in isolating genes with stable expression. The intersection of the genes identified through ASC with commonly used reference genes from the literature suggests that actin and ubiquitin ligase may be useful reference genes for T. pseudonana and potentially other diatoms. With the wealth of transcriptome sequence data becoming available, ASC can be easily applied to transcriptome datasets from other phytoplankton to identify reference genes.

Description: This research was funded by the National Science Foundation grant #OCE-0723667 (to Sonya T. Dyhrman, Mak A. Saito, Bethany D. Jenkins, and Tatiana A. Rynearson). Harriet Alexander is funded under a National Defense Science and Engineering Graduate (NDSEG) Fellowship.

Keywords: Thalassiosira pseudonana ; Diatom ; Phytoplankton ; Housekeeping genes ; RT-qPCR ; Transcriptome ; Relative gene expression ; Reference gene