EEOB publications December 1 - December 31

January 17, 2023

EEOB publications December 1 - December 31

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Never, Ever Make an Enemy… Out of an Anemone: Transcriptomic Comparison of Clownfish Hosting Sea Anemone Venoms

Alonso Delgado, Charlotte Benedict, Jason Macrander and Marymegan Daly. Mar. Drugs 2022, 20(12), 730;


Sea anemones are predatory marine invertebrates and have diverse venom arsenals. Venom is integral to their biology, and is used in competition, defense, and feeding. Three lineages of sea anemones are known to have independently evolved symbiotic relationships with clownfish, however the evolutionary impact of this relationship on the venom composition of the host is still unknown. Here, we investigate the potential of this symbiotic relationship to shape the venom profiles of the sea anemones that host clownfish. We use transcriptomic data to identify differences and similarities in venom profiles of six sea anemone species, representing the three known clades of clownfish-hosting sea anemones. We recovered 1121 transcripts matching verified toxins across all species, and show that hemolytic and hemorrhagic toxins are consistently the most dominant and diverse toxins across all species examined. These results are consistent with the known biology of sea anemones, provide foundational data on venom diversity of these species, and allow for a review of existing hierarchical structures in venomic studies.

Flower morphology variation in five species of Penstemon (Plantaginaceae) displaying Hymenoptera pollination syndrome

Rosa A. Rodriguez-Peña, Andrea D. Wolfe. STRUCTURAL BOTANY / BOTÁNICA ESTRUCTURAL.  Botanical Sciences, 101(1), 217-232.


Background: Geographic distance promotes phenotypic variation by facilitating environmental distance, limiting gene flow, and exposing plants to different pollen vectors. Therefore, understanding how plant morphology changes across a geographic range improves our understanding of the drivers of morphological diversification both on a macro- and micro-evolutionary scale.

Questions: 1) How do geographic location and abiotic factors affect flower morphology between populations? 2) Is there a geographic pattern of flower morphology variation? and 3) How does yearly variation in temperature and precipitation affect flower morphology within populations?

Studied species: Penstemon albidus, P. fruticosus, P. glandulosus, P. speciosus, and P. whippleanus

Study site and dates: The continental USA, summers of 2017 and 2018

Methods: Fifty-seven populations and 496 individuals were selected at random to measure ten floral traits. Bioclimatic variables were extracted from the WorldClim database and NOAA. Linear models, partial least squares regression, Mantel tests and canonical correlation analysis were used to analyze the data.

Results: Geographic variables alone explained a significant portion of the variation in flower morphology in two species, while in others, flower morphology did not vary despite large geographic distances. Penstemon albidus and P. whippleanus flowers increase in size from south-north, while P. glandulosus and P. speciosus exhibited an east-west increasing trend. Additionally, mean annual precipitation was the most important variable influencing P. glandulosusflower morphology.

Conclusions: Geographic distance facilitates isolation-by-distance and isolation-by-phenology as well as abiotic differences between populations; however, other factors such as pollinators might be keeping populations morphologically homogeneous despite large geographic distance.

The impacts of fine-tuning, phylogenetic distance, and sample size on big-data bioacoustics

Kaiya L. Provost , Jiaying Yang, Bryan C. Carstens. 2022. PLoS ONE 17(12): e0278522.


Vocalizations in animals, particularly birds, are critically important behaviors that influence their reproductive fitness. While recordings of bioacoustic data have been captured and stored in collections for decades, the automated extraction of data from these recordings has only recently been facilitated by artificial intelligence methods. These have yet to be evaluated with respect to accuracy of different automation strategies and features. Here, we use a recently published machine learning framework to extract syllables from ten bird species ranging in their phylogenetic relatedness from 1 to 85 million years, to compare how phylogenetic relatedness influences accuracy. We also evaluate the utility of applying trained models to novel species. Our results indicate that model performance is best on conspecifics, with accuracy progressively decreasing as phylogenetic distance increases between taxa. However, we also find that the application of models trained on multiple distantly related species can improve the overall accuracy to levels near that of training and analyzing a model on the same species. When planning big-data bioacoustics studies, care must be taken in sample design to maximize sample size and minimize human labor without sacrificing accuracy.

Inferring population connectivity in eastern massasauga rattlesnakes (Sistrurus catenatus) using landscape genetics

Scott A. Martin, William E. Peterman, Gregory J. Lipps Jr., H. Lisle Gibbs. 2022. Ecological Applications.


Assessing the environmental factors that influence the ability of a threatened species to move through a landscape can be used to identify conservation actions that connect isolated populations. However, direct observations of species' movement are often limited, making the development of alternate approaches necessary. Here we use landscape genetic analyses to assess the impact of landscape features on the movement of individuals between local populations of a threatened snake, the eastern massasauga rattlesnake (Sistrurus catenatus). We linked connectivity data with habitat information from two landscapes of similar size: a large region of unfragmented habitat and a previously studied fragmented landscape consisting of isolated patches of habitat. We used this analysis to identify features of the landscape where modification or acquisition would enhance population connectivity in the fragmented region. We found evidence that current connectivity was impacted by both contemporary land-cover features, especially roads, and inherent landscape features such as elevation. Next, we derived estimates of expected movement ability using a recently developed pedigree-based approach and least-cost paths through the unfragmented landscape. We then used our pedigree and resistance map to estimate resistance polygons of the potential extent for S. catenatus movement in the fragmented landscape. These polygons identify possible sites for future corridors connecting currently isolated populations in this landscape by linking the impact of future habitat modification or land acquisition to dispersal ability in this species. Overall, our study shows how modeling landscape resistance across differently fragmented landscapes can identify habitat features that affect contemporary movement in threatened species in fragmented landscapes and how this information can be used to guide mitigation actions whose goal is to connect isolated populations.

Testing hypotheses of hybrid taxon formation in the shrubby beardtongues (Penstemon subgenus Dasanthera)

Benjamin W. Stone, Rosa A. Rodríguez-Peña, Andrea D. Wolfe. 2022.



Hybridization is increasingly being identified in the genomes of species across the tree of life, leading to a general recognition that hybridization plays an important role in the generation of species diversity. While hybridization may increase species diversity directly via the formation of new taxa through hybrid speciation, it may also act indirectly via the exchange of phenotypic and genetic variance between species, which may in turn stimulate future speciation events.


Using high-throughput sequence data, we resolved phylogenetic relationships and investigated the role of hybridization as a diversification mechanism in the shrubby beardtongues (Penstemon subgenus Dasanthera), a group of North American wildflowers that has undergone a recent and rapid adaptive radiation. Specifically, we tested four hypotheses of hybrid taxon formation resulting from hybridization between P. davidsonii and P. fruticosus.


Species tree inference supports the monophyly of subgenus Dasanthera and elucidates relationships between taxa distributed in the Cascades and Sierra Nevada Mountains. Results also provide evidence of gene flow between P. davidsonii and P. fruticosus and support at least one hybrid origin hypothesis (P. davidsonii var. menziesii) in a region of contemporary distributional overlap. Hybridization may have also been facilitated by historical overlap in geographic distribution caused by species' responses to climatic changes during the Pleistocene.


Our results support a history of hybridization between focal taxa in a rapidly radiating clade of plants and more broadly contribute to our growing understanding of the role of hybridization as a diversification mechanism in plants.

Evolution of weedy giant ragweed (Ambrosia trifida): Multiple origins and gene expression variability facilitates weediness

Bo Li, Andrea R. Gschwend, Stephen M. Hovick, Amanda Gutek, Leah McHale, S. Kent Harrison. 2022. Ecology and Evolution. DOI: 10.1002/ece3.9590


Agricultural weeds may originate from wild populations, but the origination patterns and genetics underlying this transition remain largely unknown. Analysis of weedy- wild paired populations from independent locations may provide evidence to identify key genetic variation contributing to this adaptive shift. We performed genetic vari- ation and expression analyses on transcriptome data from 67 giant ragweed samples collected from different locations in Ohio, Iowa, and Minnesota and found geographi- cally separated weedy populations likely originated independently from their adja- cent wild populations, but subsequent spreading of weedy populations also occurred locally. By using eight closely related weedy-wild paired populations, we identified thousands of unique transcripts in weedy populations that reflect shared or specific functions corresponding, respectively, to both convergently evolved and population- specific weediness processes. In addition, differential expression of specific groups of genes was detected between weedy and wild giant ragweed populations using gene expression diversity and gene co-expression network analyses. Our study suggests an integrated route of weedy giant ragweed origination, consisting of independent origination combined with the subsequent spreading of certain weedy populations, and provides several lines of evidence to support the hypothesis that gene expression variability plays a key role in the evolution of weedy species.

Agricultural conservation practices could help offset climate change impacts on cyanobacterial harmful algal blooms in Lake Erie

Michael E. Fraker, Noel R.Aloysius, Jay F.Martin, S. Conor Keitzer, David A.Dippold, Haw Yen, Jeffrey G. Arnold, Prasad Daggupati, Mari-Vaughn V. Johnson, Dale M. Robertson, Scott P.Sowa, Michael J. White, Stuart A. Ludsin. 2022. Journal of Great Lakes Research.


Harmful algal blooms (HABs) are a recurring problem in many temperate large lake and coastal marine ecosystems, caused mainly by anthropogenic eutrophication. Implementation of agricultural conservation practices (ACPs) offers a means to reduce non-point source nutrient runoff and mitigate HABs. However, the effectiveness of ACPs in a changing climate remains uncertain. We used an integrated biophysical modeling approach to predict how Lake Erie cyanobacterial HAB severity (bloom biomass) may change under several climate and ACP implementation scenarios, using western Lake Erie and its largely agricultural watershed as our study system. An ensemble of general circulation model projections was used to drive spatially explicit land use and hydrology models of the Maumee River watershed, the output of which informed a predictive model of Lake Erie HAB severity. Results show that, in the absence of changes in ACPs, the frequency of severe HABs is projected to increase during coming decades, owing to increased inputs of nutrients from the watershed. These anticipated increases are due to increased total precipitation and more frequent higher-magnitude rainfall events. While further implementation of ACPs appears capable of reducing severe HAB events, widespread implementation would be necessary to reduce HAB severity below current management targets. This study highlights how continued climate change will only exacerbate the need for land management practices that can reduce nutrient runoff in agriculturally dominated ecosystems, such as Lake Erie. It also shows how interdisciplinary, biophysical modeling approaches can help identify strategies to mitigate HABs in the face of anthropogenic stressors.

Effects of activation and assisted reproduction techniques on the composition, structure, and properties of the sauger (Sander Canadensis) spermatozoa plasma membrane

Bryan Blawut, Barbara Wolfe, Chris Premanandan, Gustavo Schuenemann, Stuart A. Ludsin, Shan-Lu Liu, D.N. Rao Veeramachanen, Marco A. Coutinho da Silva. 2022.


The sperm plasma membrane is a multifunctional organelle essential to fertilization. However, assisted reproduction techniques often negatively affect this structure, resulting in reduced fertility. These reductions have been attributed to plasma membrane damage in a wide array of species, including fish. Considerable research has been conducted on the fish sperm membrane, but few have examined the effect of cryopreservation and other assisted reproduction techniques (ARTs) on not only membrane composition, but also specific characteristics (e.g., fluidity) and organization (e.g., lipid rafts). Herein, we determined the effects of three ARTs (testicular harvest, strip spawning, and cryopreservation) on the sperm plasma membrane, using Sauger (Sander canadensis) sperm as a model. To this end, a combination of fluorescent dyes (e.g., merocyanine 540, filipin III, cholera toxin subunit β), liquid chromatography – mass spectroscopy (LC-MS) analysis of membrane lipids, and membrane ultracentrifugation coupled with plate assays and immunofluorescence were used to describe and compare sperm fluidity, membrane composition, as well as lipid raft composition and distribution among sperm types. Stripped sperm became more fluid following motility activation (40% increase in highly fluid cells characterized by a 2 × increase in fluorescence) and contained lipid rafts restricted to the midpiece. Testicular harvest yielded sperm with characteristics similar to stripped sperm. By contrast, cryopreservation impacted every aspect of membrane physiology. Two cell populations, one highly fluid and the other rigid, resulted from the freeze-thaw process. Cryopreservation reduced lipid raft cholesterol content by 44% and flotilin-2 (a lipid raft marker) was partially displaced owing to a decrease in buoyancy. Unlike stripped and testicular sperm, LC-MS analysis revealed increases in oxidative damage markers, membrane destabilization, and apoptotic signaling in cryopreserved sperm. Ultrastructural analysis also revealed widespread physical damage to the membrane following freeze-thaw. Sperm motility, however, was unrelated to any measure of membrane physiology used in this study. Our results demonstrate that ARTs have the potential to substantially affect the sperm plasma membrane, but not always detrimentally. These results provide multiple potential biomarkers of sperm quality as well as insight into sources of sub-fertility resulting from use of ARTs.

Watershed- and reach-scale drivers of phosphorus retention and release by streambed sediment in a western Lake Erie watershed during summer

Rebecca M.Kreiling, Patrik M.Perner, Kenna J.Breckner, Tanja N.Williamson, Lynn A.Bartsch, James M.Hood, Nathan F.Manning, Laura T.Johnson. Science of The Total Environment. Volume 863, 10 March 2023, 160804.


Reducing phosphorus (P) concentrations in aquatic ecosystems, is necessary to improve water quality and reduce the occurrence of harmful cyanobacterial algal blooms. Managing P reduction requires information on the role rivers play in P transport from land to downstream water bodies, but we have a poor understanding of when and where river systems are P sources or sinks. During the summers of 2019 and 2021, we sampled streambed sediment at 78 sites throughout the Maumee River network (a major source of P loads to Lake Erie) focusing on the zero equilibrium P concentration (EPC0), the soluble reactive phosphorus (SRP) concentration at which sediment neither sorbs nor desorbs P. We used structural equation modeling to identify direct and indirect drivers of EPC0. Stream sediment was a P sink at 40 % and 67 % of sites in 2019 and 2021, respectively. During both years, spatial variation in EPC0 was shaped by stream water SRP concentrations, sediment P saturation, and sediment physicochemical characteristics. In turn, SRP concentrations and sediment P saturation (PSR) were influenced by agricultural land use and stream size. Effect of stream size differed among years with stream size having a greater effect on SRP in 2019 and on PSR in 2021. Streambed sediment is currently a net P sink across the sites sampled in the Maumee River network during summer, but sediment at these locations, especially sites in headwater streams, may become a P source if stream water SRP concentrations decrease. Our results improve the understanding of watershed- and reach-scale controls on EPC0 but also indicate the need for further research on how changes in SRP concentration as a result of conservation management implementation influences the role of streambed sediment in P transport to Lake Erie.