Linking fluvial barriers and habitat specialization in the evolution of microendemism in riverine fishes
Data files
Jun 30, 2026 version files 506.77 MB
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BPP_4sp.ctl
1.42 KB
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BPP_5sp.ctl
1.45 KB
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BPP_loci.csv
7.08 MB
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EnvironmentalVariableAnalysis.R
12.11 KB
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Eth_bellator_chermocki_background_variables.csv
528.61 KB
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Eth_bellator_presence_variables_obscured.csv
5.74 KB
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Eth_brevirostrum_background_variables.csv
756.17 KB
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Eth_brevirostrum_presence_variables_obscured.csv
8.76 KB
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Eth_chermocki_presence_variables_obscured.csv
1.54 KB
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Kim_etal_IQTREE2_mltree.treefile
10.99 KB
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Kim_etal_PoMo_data.cf
2.18 MB
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Kim_etal_PoMo_SpeciesTree.treefile
3.94 KB
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Kim_etal_SNAPP_data.nex
96.42 KB
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Kim_etal_SNAPP_TimeCalibratedSpeciesTree.nex
18.57 KB
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Kim_etal_SVDQuartets_data.nex
5.63 MB
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Kim_etal_SVDQuartets_SpeciesTree.nex
3.78 KB
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Not_chlorobranchius_background_variables.csv
1.04 MB
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Not_chlorobranchius_presence_variables.csv
8.57 KB
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params-iPyrad.txt
3.02 KB
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README.md
12.68 KB
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topo_analysis.zip
312.04 MB
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Treemix.snps.hdf5
177.33 MB
Abstract
Species with highly restricted geographic distributions are key components of biodiversity hotspots. In southeastern North America, a global hotspot of temperate freshwater biodiversity, such microendemic distributions exhibited by freshwater fishes have been attributed to geographic isolation related to spatial shifts in suitable habitat associated with climatic and geologic processes. Here, through examination of phylogenetic relationships and analysis of abiotic characteristics of the environment, we test hypotheses for the origin of microendemism within the Holiday Darter Etheostoma brevirostrum (Suttkus and Etnier) species complex. Phylogenomic analyses of double digest restriction-site associated DNA (ddRAD) sequence data find four geographically disjunct and highly diverged lineages, each of which exhibits remarkably narrow geographic distributions in the Coosa River system in Alabama, Georgia, and Tennessee, USA. We employ comparative analyses of environmental variables to quantify the degree of ecological specialization exhibited by E. brevirostrum as compared to other darter lineages that have also diversified at small scales within other river basins. We propose that ecological specialization to high-elevation streams underlain by metamorphic rock as well as the presence of knickpoints, steep sections of river channels that act as barriers to dispersal between tributaries, generated opportunities for geographic isolation and diversification in E. brevirostrum.
The files in this repository include datasets and scripts that can reproduce analyses from the related manuscript.
Dataset DOI: 10.5061/dryad.4qrfj6qkk
Description of the data and file structure
The dataset includes data and scripts to reproduce the analyses in Stokes, Kim, et al., 2026 Evolution. In this project, we analyze double-digest restriction-site associated DNA sequence data (archived at the NCBI archive, BioProject Accession Number PRJNA1163742). We perform phylogenetic and population genomic analyses. We then analyze abiotic variables to test for evidence of ecological specialization.
Genomic Analyses
The files below were used for phylogenomic analyses of Etheostoma brevirostrum including inference of a maximum-likelihood phylogeny (IQTREE), a polymorphism-aware species tree (PoMo), a time-calibrated tree (SNAPP), a species-tree inferred using SVDQuartets, gene-flow anlaysis using TreeMix, files for estimating the genealogical divergence index (gdi) using BPP, and a parameter file for the processing of the ddRAD sequence data using iPyrad.
File: Kim_etal_IQTREE2_mltree.treefile
Description: Results of the maximum likelihood phylogenetic analysis of concatenated ddRAD-loci data with IQTREE2.
File: Kim_etal_PoMo_SpeciesTree.treefile
Description: Results of the species tree analysis with the Polymorphism-aware Phylogenetic Model (PoMo)
File: Kim_etal_SNAPP_TimeCalibratedSpeciesTree.nex
Description: Results of the divergence time analysis with SNAPP
File: Kim_etal_SNAPP_data.nex
Description: Input data used in the divergence time analysis with SNAPP
File: Kim_etal_PoMo_data.cf
Description: Input data used in the PoMo analysis
File: Kim_etal_SVDQuartets_SpeciesTree.nex
Description: Results of the species tree analysis with SVDQuartets
File: Kim_etal_SVDQuartets_data.nex
Description: Input data used in the species tree analysis with SVDQuartets
File: BPP_4sp.ctl
Description: Input file for 4-species BPP analysis.
File: BPP_5sp.ctl
Description: Input file for 5-species BPP analysis.
File: params-iPyrad.txt
Description: Parameter file for iPyrad data processing.
File: BPP_loci.csv
Description: Data file for BPP analysis.
File: Treemix.snps.hdf5
Description: Data file for TreeMix analysis.
Environmental Niche Modeling
The files below were used for environmental niche modeling. There are six data files (.csv) that contain environmental variable data for each of the species analyzed. There is also an R script that can be used with these files to infer the relative degree of specialization in these three species, as well as evidence for niche conservatism/divergence within Etheostoma brevirostrum.
The data in these files were compiled from a variety of sources:
- EPA StreamCat (https://www.epa.gov/national-aquatic-resource-surveys/streamcat-dataset)
- National Land Cover NLCD database
- Fishnet2 (https://www.fishnet2.net/
- State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- National Hydrography Dataset v2 (https://www.usgs.gov/national-hydrography/national-hydrography-dataset)
- Hydrosheds hydrologically conditioned North America digital elevation model (3-arc second) (https://www.hydrosheds.org/products)
- National Elevation Dataset USGS 3DEP program, acquired from https://opentopography.org/
Files: [Species]_background/presence_variables.csv
- Eth_bellator_chermocki_background_variables.csv
- Eth_bellator_presence_variables_obscured.csv
- Eth_brevirostrum_background_variables.csv
- Eth_brevirostrum_presence_variables_obscured.csv
- Eth_chermocki_presence_variables_obscured.csv
- Not_chlorobranchius_background_variables.csv
- Not_chlorobranchius_presence_variables.csv
Description: Data files that contain environmental variable data for each species complex (Nothonotus chlorobranchius, Etheostoma brevirostrum, and Etheostoma bellator and congeners).
Note: If file contains "_obscured" in the filename, the latitude and longitude have been obscured because of the vulnerable status of the species. The latitude and longitude are reported to one decimal degree of accuracy.
Variables
Note: Some rock-type and population variables are not present in all files.
- latitude: location (decimal degrees)
- longitude: location (decimal degrees)
- comid: Unique stream reach identifier for the National Hydrography Dataset
- hydrlcondcat: Hydraulic conductivity (%)
- tmean8110cat: Mean Annual Temperature (Degrees Celcius)
- precip8110cat: Mean Annual Precipitation (mm/yr)
- slope: topographic gradient (m/m)
- qe_ma: mean annual discharge (m3 /s)
- ve_ma: mean velocity (m/s)
- meanelevsmo: mean elevation (m)
- Forest: % of HUC-12 catchment covered by the NLCD landcover category
- Developed: % of HUC-12 catchment covered by the NLCD landcover category
- BAH: not used
- Sedimentary_Clastic: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Unconsolidated_Coarse_detrital: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Sedimentary_Carbonate: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Metamorphic_Metasedimentary: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Metamorphic_Schist: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Sedimentary_Chemical: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Unconsolidated_Coarse_detrital: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Metamorphic_Amphibolite: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Metamorphic_Gneiss: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Igneous_Plutonic: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Metamorphic_Metaigneous: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Igneous_Plutonic: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Population: text describing which lineage they are assigned to (as inferred from the phylogenomic analyses)
- Population2:text describing which lineage they are assigned to (here we split the Etowah into the Etowah and Amicalola Rivers)
- Metamorphic_Metaigneous: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Tectonite: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- Igneous_Volcanic: % of HUC-12 catchment covered by the rock-type as specified by MajorRockType in the State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
File: EnvironmentalVariableAnalysis.R
Description: Script for analysis of environmental variables.
Topographic Analysis
The files and scripts here were used to make topographic maps of the study area, geologic maps of the study area, plot longitudinal topographic profiles of the rivers of interest, and identify major knickpoints that separate lineages of Etheostoma brevirostrum. We use the knickpointfinder algorithm of TopoToolbox2 in MATLAB (https://topotoolbox.wordpress.com/download/)
File: topo_analysis.zip
Description: Scripts and files necessary for topographic analysis.
File: coos_geoproj.tif
Description: geologic map
File: CoosaDEM_clip.tif
Description: digital elevation model
File: E_brevirostrum_localites_obscured.xlsx
Description: E. brevirostrum localities used in making maps. Note that latitude and longitude are obscured to one decimal degree in the file.
File: faults_proj.shp
Description: vector file of fault locations that separate Blue Ridge from Valley and Ridge
File: topoanalysis.m
Description: MATLAB script for conducting topographic analysis (mapping and identifying knickpoint)
Code/software
ipyrad v.09.90
TreeMix v.1.13( implemented in ipyrad)
IQ-TREE2
PoMo (implemented in IQ-TREE2)
ModelFinder (implemented in IQ-TREE2)
FigTree v. 1.3.1
PoMo
BPP v.4.1
SVDQuartets (implemented in PAUP* v.4.0a168)
SNAPP (implemented in BEAST v.2.5.2)
TreeAnnotator (implemented in BEAST v.2.5.2)
VCFtools v.0.1.16
Tracer v.1.7.1
R packages: LEA, PopGenome, Hierfstat, StreamCatTools, FactoMineR
MATLAB
TopoToolbox2
ArcGIS Pro
Access information
Other publicly accessible locations of the data:
- DNA sequence data is available at the NCBI archive - BioProject Accession Number PRJNA1163742.
- A reference genome was used: Etheostoma perlongum NCBI Assembly No. GCA_026937815.1
Environmental niche analysis and topographic analysis used data from the following sources:
- EPA StreamCat (https://www.epa.gov/national-aquatic-resource-surveys/streamcat-dataset)
- National Land Cover NLCD database
- Fishnet2 (https://www.fishnet2.net/
- State Geologic Map Compilation (https://www.sciencebase.gov/catalog/item/5888bf4fe4b05ccb964bab9d)
- National Hydrography Dataset v2 (https://www.usgs.gov/national-hydrography/national-hydrography-dataset)
- Hydrosheds hydrologically conditioned North America digital elevation model (3-arc second) (https://www.hydrosheds.org/products)
- National Elevation Dataset USGS 3DEP program, acquired from https://opentopography.org/
