Development and evaluation of 200 novel SNP assays for population genetic studies of westslope cutthroat trout and genetic identification of related taxa

Abstract

DNA sequence data were collected and screened for single nucleotide polymorphisms (SNPs) in westslope cutthroat trout (Oncorhynchus clarki lewisi) and also for substitutions that could be used to genetically discriminate rainbow trout (O. mykiss) and cutthroat trout, as well as several cutthroat trout subspecies. In total, 260 expressed sequence tag-derived loci were sequenced and allelic discrimination genotyping assays developed from 217 of the variable sites. Another 50 putative SNPs in westslope cutthroat trout were identified by restriction-site-associated DNA sequencing, and seven of these were developed into assays. Twelve O. mykiss SNP assays that were variable within westslope cutthroat trout and 12 previously published SNP assays were also included in downstream testing. A total of 241 assays were tested on six westslope cutthroat trout populations (N = 32 per population), as well as collections of four other cutthroat trout subspecies and a population of rainbow trout. All assays were evaluated for reliability and deviation from Hardy-Weinberg and linkage equilibria. Poorly performing and duplicate assays were removed from the data set, and the remaining 200 assays were used in tests of population differentiation. The remaining markers easily distinguished the various subspecies tested, as evidenced by mean G(ST) of 0.74. A smaller subset of the markers (N = 86; average G(ST) = 0.40) was useful for distinguishing the six populations of westslope cutthroat trout. This study increases by an order of magnitude the number of genetic markers available for the study of westslope cutthroat trout and closely related taxa and includes many markers in genes (developed from ESTs).

Development and Validation of Single Nucleotide Polymorphisms (SNPs) Markers from Two Transcriptome 454-Runs of Turbot (Scophthalmus maximus) Using High-Throughput Genotyping

“…The samples from the Cantabrian turbot population were globally in accordance with HWE expectations when tested simultaneously for all loci ( p = 0.9999). These polymorphic values were in the range to those previously described in the species [29], and they were also similar to those reported in other fish species [42,44]. No Linkage disequilibrium (LD) was detected among the 6328 loci pairs after Bonferroni correction ( p = 0.0004).…”

“…One hundred and thirteen SNPs of the 140 tested were amplified and genotyped, 111 being polymorphic in a wild Cantabrian population, showing the utility of the new NGS techniques for true SNP detection (conversion rate = 79.3%). Diversity levels at the population were similar to previous studies [29,42,44] and were in accordance with HWE expectations. An important number of these polymorphic SNPs were located in the coding region and 16 of them (14.4%) represented non-synonymous changes at genes related to immune response and gonad differentiation processes as shown by the detected GO terms.…”

Relative contributions of neutral and non‐neutral genetic differentiation to inform conservation of steelhead trout across highly variable landscapes

“…; Campbell et al. ), we found no evidence for multiple evolutionary lineages in the Snake River, but local environments may influence their differentiation. For reference, six Snake River hatchery stocks were genetically similar to their natural‐origin counterparts that distinguish the Snake River DPS.…”

Environmental DNA in lake sediment reveals biogeography of native genetic diversity

“…; Campbell et al . ). These three species were chosen because they have well‐documented histories of human‐mediated introduction across the landscape and have hybridized extensively throughout North America.…”