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Reevaluating claims of ecological speciation in Halichoeres bivittatus

Citation

Warren, Dan et al. (2021), Reevaluating claims of ecological speciation in Halichoeres bivittatus, Dryad, Dataset, https://doi.org/10.5061/dryad.fj6q573vf

Abstract

Allopatry has traditionally been viewed as the primary driver of speciation in marine taxa, but the geography of the marine environment and the larval dispersal capabilities of many marine organisms render this view somewhat questionable. In marine fishes, one of the earliest and most highly cited empirical examples of ecological speciation with gene flow is the slippery dick wrasse, Halichoeres bivittatus. Evidence for this cryptic or incipient speciation event was primarily in the form of a deep divergence in a single mitochondrial locus between the northern and southern Gulf of Mexico, combined with a finding that these two haplotypes were associated with different habitat types (“tropical” vs. “subtropical”) in the Florida Keys and Bermuda, where they overlap. Here we examine habitat assortment in the Florida Keys using a broader sampling of populations and habitat types than were available for the original study. We find no evidence to support the claim that haplotype frequencies differ between habitat types, and little evidence to support any differences between populations in the Keys. These results undermine claims of ecological speciation with gene flow in Halichoeres bivittatus. Future claims of this type should be supported by multiple lines of evidence that illuminate potential mechanisms and allow researchers to rule out alternative explanations for spatial patterns of genetic differences.

Methods

We sampled eight populations in the Florida Keys including four populations on the edge of the continental shelf (Sombrero Light, 11 Foot Mound, XMuta, and Tennessee Reef), two populations on patch reefs in the inshore channel (East Washerwoman and East Turtle Shoal), and two grass beds located directly offshore in water < 2m in depth (near mile marker 62 on Long Key and behind Keys Marine Lab (KML) on Vaca Key).  For broader geographic context we also sampled fishes from two sites further north on the gulf coast of Florida, two sites in the Bahamas, and one site from Belize.  Florida and Bahamas specimens were collected in 2005 and 2006, and Belize specimens were collected in 2006.  In addition to comparing fore reef and inshore patch reef, we included the grass bed habitat as it experiences even greater seasonal and diurnal fluctuations in temperature than the inshore patch reef and as such provides an additional test of the proposed habitat segregation.

All animal handling procedures were approved by the University of California, Davis Institutional Animal Care and Use Committee.  Fish were caught using a combination of hand nets, barrier nets, and otter trawls.  Specimens were euthanized using MS-222 dissolved in seawater, and samples were taken from muscle tissue and preserved in 95% ethanol.  We extracted DNA using DNeasy™ (Qiagen) columns and PCR amplified a fragment of the  mitochondrial cytochrome B gene using the L14768 and H15496 primers from Rocha et al. (2005).  PCR products were cleaned using ExoSap-IT (USB Corp.). Purified templates were dye labeled using BigDye (ABI) and sequenced on an ABI 3077 automated DNA Sanger sequencer.  Sequences were aligned using ClustalW (Thompson, Gibson, and Higgins 2002).

Rocha, Luiz A., D. Ross Robertson, Joe Roman, and Brian W. Bowen. 2005. “Ecological Speciation in Tropical Reef Fishes.” Proceedings. Biological Sciences / The Royal Society 272 (1563): 573–79.

Thompson, Julie D., Toby J. Gibson, and Des G. Higgins. 2002. “Multiple Sequence Alignment Using ClustalW and ClustalX.” Current Protocols in Bioinformatics / Editoral Board, Andreas D. Baxevanis ... [et Al.] Chapter 2 (August): Unit 2.3.