Contributors | Affiliation | Role |
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Lee, Carol E. | University of Wisconsin (UW-Madison) | Principal Investigator, Contact |
Diaz, Juanita A. | University of Wisconsin (UW-Madison) | Scientist, Contact |
Stern, David B. | University of Wisconsin (UW-Madison) | Scientist |
Newman, Sawyer | Woods Hole Oceanographic Institution (WHOI BCO-DMO) | BCO-DMO Data Manager |
These data have been deposited in NCBI under BioProject number PRJNA923656.
Methods and Sampling:
Wild Eurytemora affinis populations were collected from three locations in the North Sea using bongo nets with 100 micrometer (μm) mesh. Copepods were stored in RNAlater. Sampling locations included two fresh water lakes and one brackish estuary. From each population, individual copepods (100 individuals, 50:50 male:female) were pooled and their DNA was extracted using the DNeasy Blood and Tissue Extraction kit (Qiagen, Inc.). Paired-end whole-genome sequencing libraries were prepared using the Illumina Nextera DNA kit (Illumina, Inc.) and sequenced on three lanes of an Illumina HiSeq 2000 sequencer at the Maryland Institute of Genome Science, generating an average of approximately 158 million paired-end (100 bp) reads per pool.
Processing notes from researcher:
Raw sequence reads were mapped to a reference genome to call SNPs. We then detected SNPs with significant signatures of selection across temperature and salinity gradients. Signatures of selection were detected using an outlier approach and an association with an environmental variable (i.e., temperature or salinity) approach.
The following software was used:
BayPass v.2.2, BEDOPS v.2.4.39, BITE v.1.2.8, BLAST 2.7.1+, BWA-MEM v.0.7.17, GATK v.3.8, Gowinda v.1.12, ggplot2 v.3.3.5, gplots v.3.1.3, OptM v.0.1.6, PHYLIP v.3.697, Picard v. 2.18.27, poolfstat v.1.0, poolfstat v.2.1.1, PoPoolation v.1.2.2, PoPoolation2 v.1.201, SAMtools v.1.3.1, stats v.4.1.2, TreeMix v.1.13, Trimmomatic v.0.39, VarScan v.2.4.3, vegan v.2.6-2, https://github.com/TheDBStern/Baltic_Lab_Wild (DOI:10.5281/zenodo.6615047), https://github.com/juanitadiaz/Europe_Wild_LocalAdaptation_GeneFlow (DOI:10.5281/zenodo.7819560), https://github.com/carolindahms/TreeMix
BCO-DMO Processing Notes:
Parameter | Description | Units |
Location | Location of sample site | unitless |
Collection_Date | Date of sample collection in format YYYY-MM-DD | unitless |
Sample_Code | Unique identifier for sample | unitless |
Sample_Salinity | Sea surface salnity measured by refractometer | Practical salinity units (PSU) |
Sample_Temperature | Sea surface temperature measured by thermometer | degrees Celsius |
Latitude | Latitude of collection location in decimal degrees. A positive value indicates a Northern coordinate. | decimal degrees |
Longitude | Longitude of collection location in decimal degrees. A positive value indicates an Eastern coordinate. | decimal degrees |
BioSample | NCBI BioSample ID | unitless |
SRA_Run | NCBI SRA Run number | unitless |
Dataset-specific Instrument Name | Illumina HiSeq 2000 sequencer |
Generic Instrument Name | Automated DNA Sequencer |
Dataset-specific Description | Used to sequence pooled E. affinis samples. |
Generic Instrument Description | General term for a laboratory instrument used for deciphering the order of bases in a strand of DNA. Sanger sequencers detect fluorescence from different dyes that are used to identify the A, C, G, and T extension reactions. Contemporary or Pyrosequencer methods are based on detecting the activity of DNA polymerase (a DNA synthesizing enzyme) with another chemoluminescent enzyme. Essentially, the method allows sequencing of a single strand of DNA by synthesizing the complementary strand along it, one base pair at a time, and detecting which base was actually added at each step. |
Dataset-specific Instrument Name | Bongo net |
Generic Instrument Name | Bongo Net |
Dataset-specific Description | E. affinis samples were collected using a Bongo net with 100 μm mesh. |
Generic Instrument Description | A Bongo Net consists of paired plankton nets, typically with a 60 cm diameter mouth opening and varying mesh sizes, 10 to 1000 micron. The Bongo Frame was designed by the National Marine Fisheries Service for use in the MARMAP program. It consists of two cylindrical collars connected with a yoke so that replicate samples are collected at the same time. Variations in models are designed for either vertical hauls (OI-2500 = NMFS Pairovet-Style, MARMAP Bongo, CalVET) or both oblique and vertical hauls (Aquatic Research). The OI-1200 has an opening and closing mechanism that allows discrete "known-depth" sampling. This model is large enough to filter water at the rate of 47.5 m3/minute when towing at a speed of two knots. More information: Ocean Instruments, Aquatic Research, Sea-Gear |
NSF Award Abstract:
Drastic changes in the global water cycle and increases in ice melt are causing the freshening of Northern coastal seas. The combination of both reduced salinity and increased temperature will likely act in concert to reduce populations of estuarine and marine organisms. Data indicate that reduced salinity and high temperature would each increase the energy costs as well as reduce survival and reproduction of the common copepod Eurytemora affinis. This project will examine the joint effects of salinity reduction and temperature increase on the evolutionary responses of populations of E. affinis in the wild, as well as in selection experiments in the laboratory. This study will provide novel insights into responses of organisms to climate change, as no study has analyzed the joint impacts of salinity and temperature on evolutionary responses, and relatively few studies have examined the impacts of declining salinity. In general, how selection acts at the whole genome level is not well understood, particularly for non-model organisms. As a dominant estuarine copepod, E. affinis is among the most important species sustaining coastal food webs and fisheries in the Northern Hemisphere, such as salmon, herring, and anchovy. Thus, insights into its evolutionary responses with changing climate have important implications for sustainability of fisheries and food security. Two graduate students from historically underrepresented groups will be trained during this project. The project will have additional societal benefits, including development of educational modules for K-12 students and international collaboration.
This study will address the following questions: (1) To what extent could populations evolve in response to salinity and temperature change, and what are the fitness and physiological costs? (2) How will populations respond to the impacts of salinity-temperature interactions? (3) Do wild populations show evidence of natural selection in response to salinity and temperature? To analyze the evolutionary responses of E. affinis populations to the coupled impacts of salinity and temperature, the investigator will perform laboratory selection experiments and population genomic surveys of wild populations. Selection experiments constitute powerful tools for determining the rate, trajectory, and limits of adaptation. During laboratory selection, evolutionary shifts in fitness-related traits and genomic expression will be examined, as well as genomic signatures of selection in response to low salinity and high temperature selection regimes. The investigator will also conduct population genomic sequencing of E. affinis populations that reside along salinity and temperature gradients in the St. Lawrence and Baltic Sea, and identify genes that show signatures of selection. The project will determine whether the loci that show signatures of selection in the wild populations are the same as those favored during laboratory selection. This reproducibility will provide greater confidence that the genes involved in adaptation to salinity and/or temperature have been captured.
Funding Source | Award |
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NSF Division of Ocean Sciences (NSF OCE) |