Abstract

BackgroundComparative genomics approaches help to shed light on evolutionary processes that shape differentiation between lineages. The nine-spined stickleback (Pungitius pungitius) is a closely related species of the ecological ‘supermodel’ three-spined stickleback (Gasterosteus aculeatus). It is an emerging model system for evolutionary biology research but has garnered less attention and lacks extensive genomic resources. To expand on these resources and aid the study of sticklebacks in a phylogenetic framework, we characterized nine-spined stickleback transcriptomes from brain and liver using deep sequencing.ResultsWe obtained nearly eight thousand assembled transcripts, of which 3,091 were assigned as putative one-to-one orthologs to genes found in the three-spined stickleback. These sequences were used for evaluating overall differentiation and substitution rates between nine- and three-spined sticklebacks, and to identify genes that are putatively evolving under positive selection. The synonymous substitution rate was estimated to be 7.1 × 10-9 per site per year between the two species, and a total of 165 genes showed patterns of adaptive evolution in one or both species. A few nine-spined stickleback contigs lacked an obvious ortholog in three-spined sticklebacks but were found to match genes in other fish species, suggesting several gene losses within 13 million years since the divergence of the two stickleback species. We identified 47 SNPs in 25 different genes that differentiate pond and marine ecotypes. We also identified 468 microsatellites that could be further developed as genetic markers in nine-spined sticklebacks.ConclusionWith deep sequencing of nine-spined stickleback cDNA libraries, our study provides a significant increase in the number of gene sequences and microsatellite markers for this species, and identifies a number of genes showing patterns of adaptive evolution between nine- and three-spined sticklebacks. We also report several candidate genes that might be involved in differential adaptation between marine and freshwater nine-spined sticklebacks. This study provides a valuable resource for future studies aiming to identify candidate genes underlying ecological adaptation in this and other stickleback species.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-14-756) contains supplementary material, which is available to authorized users.

Highlights

  • Comparative genomics approaches help to shed light on evolutionary processes that shape differentiation between lineages

  • Contig assembly of the reads were combined from the four cDNA libraries into one “nine-spined stickleback transcriptome” containing 7,932 contigs ≥ 100 bp with an average coverage depth of 38 reads (Additional file 2: Table S1)

  • Functional annotation A BLASTX search returned 3,347 (42.2%) nine-spined stickleback contigs with significant hits to three-spined stickleback genes. This proportion of contigs with BLAST hits is similar to previous transcriptome studies [47,48,49], in which contigs without significant hits may consist of untranslated transcripts, chimeras or assembly artifacts

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Summary

Introduction

Comparative genomics approaches help to shed light on evolutionary processes that shape differentiation between lineages. The nine-spined stickleback (Pungitius pungitius) is a closely related species of the ecological ‘supermodel’ three-spined stickleback (Gasterosteus aculeatus) It is an emerging model system for evolutionary biology research but has garnered less attention and lacks extensive genomic resources. The three-spined stickleback (Gasterosteus aculeatus) has become a vertebrate ‘supermodel’ allowing a combination of studies at molecular, developmental, phenotypic, and population genetic levels to explore factors and processes relevant for adaptive evolution in ecologically relevant contexts [3,4]. Populations that have colonized freshwater habitats after the retreat of Pleistocene ice sheets have evolved remarkable morphological and behavioral diversity as compared to marine populations [5,6] They have repeatedly evolved changes in body shape, skeletal armor, trophic apparati, pigmentation, osmoregulatory functions, life history, and behavior [5]. Relatively little is known about the genetics of these or other traits in other stickleback species (but see: [16,17,18,19,20])

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