Evolution and conservation of Characidium sex chromosomes

Abstract

Fish species exhibit substantial variation in the degree of genetic differentiation between sex chromosome pairs, and therefore offer the opportunity to study the full range of sex chromosome evolution. We used restriction-site associated DNA sequencing (RAD-seq) to study the sex chromosomes of Characidium gomesi, a species with conspicuous heteromorphic ZW/ZZ sex chromosomes. We screened 9863 single-nucleotide polymorphisms (SNPs), corresponding to ~1 marker/100 kb distributed across the genome for sex-linked variation. With this data set, we identified 26 female-specific RAD loci, putatively located on the W chromosome, as well as 148 sex-associated SNPs showing significant differentiation (average F_ST=0.144) between males and females, and therefore in regions of more recent divergence between the Z and W chromosomes. In addition, we detected 25 RAD loci showing extreme heterozygote deficiency in females but which were in Hardy-Weinberg equilibrium in males, consistent with degeneration of the W chromosome and therefore female hemizygosity. We validated seven female-specific and two sex-associated markers in a larger sample of C. gomesi, of which three localised to the W chromosome, thereby providing useful markers for sexing wild samples. Validated markers were evaluated in other populations and species of the genus Characidium, this exploration suggesting a rapid turnover of W-specific repetitive elements. Together, our analyses point to a complex origin for the sex chromosome of C. gomesi and highlight the utility of RAD-seq for studying the composition and evolution of sex chromosomes systems in wild populations.

Figure 1

Geographic map of sampling sites for Characidium species and populations used in this study.

Figure 2

Mitotic plate of Characidium gomesi female after C-banding, showing a strong C-band positive pattern on W chromosome.

Figure 3

Distribution of genetic differentiation (F_ST) between male and female subpopulations for the 148 sex-associated SNPs (dark grey) and a random sample of 500 SNPs not associated with sex (light grey).

Figure 4

Banding patterns of validated female-specific markers identified in Characidium gomesi in Paranapanema (PR) and Tiete (TR) populations. Amplification on the W chromosome DNA library is also shown.