Abstract

The identification of genes involved in the adaptive evolution of non-model organisms with uncharacterized genomes constitutes a major challenge. This study employed a rigorous and targeted candidate gene approach to test for positive selection on protein-coding genes of the blue crab, Callinectes sapidus. Four genes with putative roles in physiological adaptation to environmental stress were chosen as candidates. A fifth gene not expected to play a role in environmental adaptation was used as a control. Large samples (n>800) of DNA sequences from C. sapidus were used in tests of selective neutrality based on sequence polymorphisms. In combination with these, sequences from the congener C. similis were used in neutrality tests based on interspecific divergence. In multiple tests, significant departures from neutral expectations and indicative of positive selection were found for the candidate gene trehalose 6-phosphate synthase (tps). These departures could not be explained by any of the historical population expansion or bottleneck scenarios that were evaluated in coalescent simulations. Evidence was also found for balancing selection at ATP-synthase subunit 9 (atps) using a maximum likelihood version of the Hudson, Kreitmen, and Aguadé test, and positive selection favoring amino acid replacements within ATP/ADP translocase (ant) was detected using the McDonald-Kreitman test. In contrast, test statistics for the control gene, ribosomal protein L12 (rpl), which presumably has experienced the same demographic effects as the candidate loci, were not significantly different from neutral expectations and could readily be explained by demographic effects. Together, these findings demonstrate the utility of the candidate gene approach for investigating adaptation at the molecular level in a marine invertebrate for which extensive genomic resources are not available.

Highlights

  • According to the neutral theory of molecular evolution, natural selection acts mainly to remove deleterious mutations from populations, leaving behind allelic variation that is almost entirely neutral [1,2]

  • Mean divergence (KJC) between C. sapidus and C. similis was highest for tps (0.054) and lowest for atps (0.003) (Table 2)

  • Thorough examination of the chromatogram sequence traces for hsp (n = 854) as well as the other four loci provided no evidence for paralogous loci or pseudogenes in the form of triple peaks

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Summary

Introduction

According to the neutral theory of molecular evolution, natural selection acts mainly to remove deleterious mutations from populations, leaving behind allelic variation that is almost entirely neutral [1,2]. The enduring value of this theory is that it predicts how genetic variation should be distributed within and between populations by non-selective forces such as gene flow and genetic drift. As the genomes of more species have been sequenced, it has become evident that natural selection acts on variation at a greater proportion of loci than anticipated by the classical neutral theory. Evidence for widespread selection has been found in the genomes of model organisms including Drosophila [3,4,5], Arabidopsis [6,7], and Homo sapiens [8,9,10]. Studies of non-model organisms have provided evidence for the importance of natural selection at the molecular level [11,12,13,14]

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