A rapid approach for sex assignment by RAD-seq using a reference genome

PLoS One. 2024 Apr 5;19(4):e0297987. doi: 10.1371/journal.pone.0297987. eCollection 2024.

Abstract

Sex identification is a common objective in molecular ecology. While many vertebrates display sexual dimorphism, determining the sex can be challenging in certain situations, such as species lacking clear sex-related phenotypic characteristics or in studies using non-invasive methods. In these cases, DNA analyses serve as valuable tools not only for sex determination but also for validating sex assignment based on phenotypic traits. In this study, we developed a bioinformatic framework for sex assignment using genomic data obtained through GBS, and having an available closely related genome assembled at the chromosome level. Our method consists of two ad hoc indexes that rely on the different properties of the mammalian heteromorphic sex chromosomes. For this purpose, we mapped RAD-seq loci to a reference genome and then obtained missingness and coverage depth values for the autosomes and X and Y chromosomes of each individual. Our methodology successfully determined the sex of 165 fur seals that had been phenotypically sexed in a previous study and 40 sea lions sampled in a non-invasive way. Additionally, we evaluated the accuracy of each index in sequences with varying average coverage depths, with Index Y proving greater reliability and robustness in assigning sex to individuals with low-depth coverage. We believe that the approach presented here can be extended to any animal taxa with known heteromorphic XY/ZW sex chromosome systems and that it can tolerate various qualities of GBS sequencing data.

MeSH terms

  • Animals
  • Genome* / genetics
  • Genomics
  • Humans
  • Mammals / genetics
  • Reproducibility of Results
  • Sex Chromosomes* / genetics
  • Y Chromosome

Grants and funding

DMP received funding from the National Geographic Society (10000-16 and 269 EC- 52880R-18) and the Sociedad Argentina para el Estudio de los Mamíferos (Premio Kravetz 2019). JIT recieved funding from the Universidad Nacional de Luján (Finalidad 3.5 año 2018). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.