Identification of Key Genes and Variants Associated with Boar Sperm Freezability Using Whole Genome Resequencing

Cryopreservation induces various cryodamages to the structural or functional aspects of boar sperm, resulting in the deterioration of sperm quality. The extent of cryodamages varies significantly among different individual boars. In our study, 50 boars with either good sperm freezability (GSF) or poor sperm freezability (PSF) were selected from a population of 402 boars. These two groups exhibited significant differences in sperm quality, acrosome integrity, mitochondrial membrane potential, and plasma membrane integrity. Subsequent whole-genome resequencing and FST analysis of GSF and PSF boars uncovered genetic differentiation among 5632 genes. Spermatogenesis, sperm structure, and lipid composition were identified as the potential factors, and 35 genes, such as RNF8, PACRG, CADM1, SPAG16, PPP3CA and MFF, were identified as key candidate genes associated with sperm freezability. Furthermore, by using MassARRAY genotyping and a general linear model, nine variants, including seven single nucleotide polymorphisms (SNPs) and two insertion variants, were found to be significantly correlated with sperm freezability. These variants, located within the candidate genes, represent potential molecular markers for sperm freezability. Our research contributes novel insights into the genetic mechanisms underlying freezability differences in boar sperm and identifies candidate genes and key molecular markers that could be applied in boar breeding practices.