Contributors | Affiliation | Role |
---|---|---|
Barott, Katie | University of Pennsylvania (Penn) | Principal Investigator |
Brown, Kristen | University of Pennsylvania (Penn) | Scientist |
Speer, Kelsey | University of Pennsylvania (Penn) | Scientist |
Glass, Benjamin | University of Pennsylvania (Penn) | Student, Contact |
Schmitt, Angela | University of Pennsylvania (Penn) | Student |
Soenen, Karen | Woods Hole Oceanographic Institution (WHOI BCO-DMO) | BCO-DMO Data Manager |
Data generated as part of a Nematostella ocean acidfication experiment published in Glass et al., 2023. (see Related Publications). Related Zenodo datasets provides further analysis and plotting of the BCO-DMO dataset here. (see Related Dataset).
Nematostella vectensis (Stephenson, 1935) anemones were collected from a salt marsh in Brigantine, New Jersey in the fall of 2020. Females were identified by inducing spawning, and 14 individuals that released eggs were chosen as the genotype pool for this experiment. Each female was then horizontally bisected through the body column using a razor blade, resulting in two genotypically identical individuals that were divided between the two experimental groups (ambient and acidic).
A clonal male population, also originating from the United States Atlantic coast, was obtained from the laboratory of Dr. Katerina Ragkousi (Amherst College) in the spring of 2021. The male population size was increased via bisection, resulting in a total of 20 genetically identical males for the experiment (N=10 per treatment).
All anemones were kept in 12 parts per thousand (ppt) artificial seawater (ASW; Instant Ocean Reef CrystalsⓇ reef salt, Spectrum Brands, Blacksburg, VA, USA) at pH 7.7–8.1 and 18°C. The animals were maintained in a dark incubator (Boekel Scientific, Feasterville-Trevose, PA, USA) and fed approximately every other day with Artemia nauplii. The experiment was performed approximately 1–1.5 years after animal collection.
Larvae from each parental treatment were individually pipetted into polymerase chain reaction (PCR) strip tubes (N=32 larvae treatment−1 temperature−1). Larvae were then exposed to one of a range of peak temperatures between 39–43°C in 0.5-degree increments. MiniAmp thermal cyclers (Thermo Fisher Scientific, Waltham, MA, USA) were used for heat ramps, which were programmed as follows: (1) 1 min at 25°C; (2) 4 min at 30°C; (3) 4 min at 38°C; (4) 1 h at the peak temperature (39–43°C); (5) 4 min at 38°C; (6) 4 min at 30°C; (7) infinite hold at 22°C.
Strip tubes containing larvae were capped, randomly assigned to positions in the thermal cyclers for the heat ramp, and then removed and uncapped as soon as the cool-down ramp was complete. Following uncapping, tubes were placed in the dark at 18°C for 48 h, then larvae were transferred with a multichannel pipette to a 96-well plate and examined for survival (no clear disintegration of tissue) under a dissecting microscope. The percentage of larvae surviving after exposure to each peak temperature was calculated as the number of larvae surviving divided by the total number of larvae exposed to each temperature.
For heat tolerance data, the percentage of larvae surviving after exposure to each peak temperature was calculated as the number of larvae surviving divided by the total number of larvae exposed to each temperature
* Adjusted parameter names to comply with database requirements
File |
---|
920813_v1_larvalheattolerance.csv (Comma Separated Values (.csv), 334 bytes) MD5:ba6e9a36f6142ce8bed1ecf18324012b Primary data file for dataset ID 920813, version 1 |
Parameter | Description | Units |
Temperature | Temperature at which larvae were treated for 1-hour as a heat shock | degrees Celsius (°C) |
Treatment | Experimental treatment into which anemones were placed (ambient or acidic seawater pH) | unitless |
Percent_surviving_larvae | The percent of larvae surivivng 48 hours after the heat shock | percentage (%) |
Dataset-specific Instrument Name | MiniAmp thermal cyclers (Thermo Fisher Scientific, Waltham, MA, USA) |
Generic Instrument Name | Thermal Cycler |
Dataset-specific Description | MiniAmp thermal cyclers (Thermo Fisher Scientific, Waltham, MA, USA) for thermal tolerance heat ramps |
Generic Instrument Description | A thermal cycler or "thermocycler" is a general term for a type of laboratory apparatus, commonly used for performing polymerase chain reaction (PCR), that is capable of repeatedly altering and maintaining specific temperatures for defined periods of time. The device has a thermal block with holes where tubes with the PCR reaction mixtures can be inserted. The cycler then raises and lowers the temperature of the block in discrete, pre-programmed steps. They can also be used to facilitate other temperature-sensitive reactions, including restriction enzyme digestion or rapid diagnostics.
(adapted from http://serc.carleton.edu/microbelife/research_methods/genomics/pcr.html) |
NSF Award Abstract:
Coral reefs are incredibly diverse ecosystems that provide food, tourism revenue, and shoreline protection for coastal communities. The ability of coral reefs to continue providing these services to society is currently threatened by climate change, which has led to increasing ocean temperatures and acidity that can lead to the death of corals, the animals that build the reef framework upon which so many species depend. This project examines how temperature and acidification stress work together to influence the future health and survival of corals. The scientists are carrying out the project in Hawaii where they have found individual corals with different sensitivities to temperature stress that are living on reefs with different environmental pH conditions. This project improves understanding of how an individual coral's history influences its response to multiple stressors and helps identify the conditions that are most likely to support resilient coral communities. The project will generate extensive biological and physicochemical data that will be made freely available. Furthermore, this project supports the education and training of undergraduate and high school students and one postdoctoral researcher in marine science and coral reef ecology. Hands-on activities for high school students are being developed into a free online educational resource.
This project compares coral responses to acidification stress in populations experiencing distinct pH dynamics (high diel variability vs. low diel variability) and with distinct thermal tolerances (historically bleaching sensitive vs. tolerant) to learn about how coral responses to these two factors differ between coral species and within populations. Experiments focus on the two dominant reef builders found at these stable and variable pH reefs: Montipora capitata and Porites compressa. Individuals of each species exhibiting different thermal sensitivities (i.e., bleached vs. pigmented) were tagged during the 2015 global coral bleaching event. This system tests the hypotheses that 1) corals living on reefs with larger diel pH fluctuations have greater resilience to acidification stress, 2) coral resilience to acidification is a plastic trait that can be promoted via acclimatization, and 3) thermally sensitive corals have reduced capacity to cope with pH stress, which is exacerbated at elevated temperatures. Coral cells isolated from colonies from each environmental and bleaching history are exposed to acute pH stress and examined for their ability to recover intracellular pH in vivo using confocal microscopy, and the expression level of proteins predicted to be involved in this recovery (e.g., proton transporters) is examined via Western blot and immunolocalization. Corals from each pH history are exposed to stable and variable seawater pH in a controlled aquarium setting to determine the level of plasticity of acidification resilience and to test for pH acclimatization in this system. Finally, corals with different levels of thermal sensitivity are exposed to thermal stress and recovery, and their ability to regulate pH is examined over time. The results of these experiments help identify reef conditions that promote coral resilience to ocean acidification against the background of increasingly common thermal stress events, while advancing mechanistic understanding of coral physiology and symbiosis.
This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
Funding Source | Award |
---|---|
NSF Division of Ocean Sciences (NSF OCE) |