Characterization of microsatellite loci for the Gulf Coast waterdog (Necturus beyeri) using paired-end Illumina shotgun sequencing and cross-amplification in other Necturus

Abstract

Amphibians are one of the most threatened groups of vertebrates (Stuart et al. 2004; Wake and Vredenburg 2008), and the application of molecular techniques to amphibian ecology and genetics has dramatically improved our ability to conserve species and populations (see Shaffer et al. [2015] for review). Microsatellites, tandem repeats of two to six nucleotides in the nuclear genome, are highly variable molecular markers that can be used to describe gene flow and genetic diversity, each of which is positively correlated with population persistence (Allendorf and Luikart 2007; Allentoft and O’Brien 2010; Avise 2004; Selkoe and Toonen 2006). Microsatellite loci have frequently been applied to studies involving terrestrial and pond breeding amphibians (Emel and Storfer 2012), but fewer studies have focused on taxa inhabiting lotic systems (Emel and Storfer 2012). For example, studies characterizing microsatellite loci are completely lacking for a group of permanently aquatic salamanders, the waterdogs and mudpuppies (Family Proteidae, Genus Necturus ) (Rafinesque 1819). The genus Necturus consists of several species of perennibranch salamanders that can be found throughout many freshwater streams, rivers, and lakes in North America (Petranka 1998). Some authorities recognize five species (Crother 2012; Petranka 1998), including the Mudpuppy ( Necturus maculosus ) (Rafinesque 1819), Gulf Coast Waterdog ( N. beyeri ) (Viosca 1937), Black Warrior Waterdog ( N. alabamensis ) (Viosca 1937), Neuse River Waterdog ( N. lewisi ) (Brimley 1924), and Dwarf Waterdog ( N. punctatus ) (Gibbes 1850). This taxonomy also recognizes two subspecies within N. maculosus , including the Common Mudpuppy ( N. m. maculosus ) and the Red River Waterdog ( N. m. louisianensis ) (Crother 2012; Petranka 1998; Schmidt 1953). Other authorities suggest that there are six or seven species within Necturus (Collins 1990; Frost 2016; Powell et al. 2016). These more diverse schemes recognize each of the aforementioned five species while also elevating the Red River Waterdog ( N. louisianensis ) (Collins 1990; Frost 2016; Powell et al. 2016; Viosca 1938) and Löding’s Waterdog ( N. lödingi or N. cf. beyeri ) (Bart et al. 1997; Guyer 2005a; Viosca 1938). Allozyme work by Guttman et al. (1990) suggests that there is at least one cryptic species of Necturus in drainages east of the Mobile Basin and south of the Alabama River, and both Bart et al. (1997) and Guyer (2005a) advise that these populations should be referred to as N. cf. beyeri . However, until range wide studies incorporating genetic and other data are published, we will follow the five species taxonomy outlined by Crother (2012) while acknowledging that certain taxa, such as N. maculosus and N. beyeri , may require systematic revision.