Abstract
Single nucleotide variations (SNVs) in the nuclear genome have been used
widely to analyze phylogenetic and population genomic structure.
Cost-effective genotyping can be achieved by sequencing PCR amplicons
using short 3-10 base sequences as primers to arbitrarily amplify
thousands of sites in the genome using only a few primers. While
previous methods have produced an insufficient number of SNVs to perform
population genomic analyses, we designed a new primer set to improve the
sequencing efficiency. To demonstrate the effectiveness of our method,
we examined the population genetic structure of the small freshwater
fish, medaka (Oryzias latipes). Specifically, we attempted to
reconstruct the genetic admixture of the orange mutant strain, Himedaka.
Although the strain is widely kept as an ornamental fish and for
experimental purposes, the genetic background of the nuclear genome of
commercial stock is less clear. We obtained 2987 informative SNVs with
no missing genotype calls for 67 individuals from 15 wild populations
and three artificial strains using the HiSeq X platform. The estimated
phylogenic and population genetic structures of the wild populations
were consistent with previous studies, corroborating the accuracy of our
genotyping method. Admixture analysis focusing on Himedaka showed that
at least two wild populations contributed SNVs to the nuclear genome of
this mutant strain. Population genomics analyses based on nuclear SNVs
data are indispensable to identify admixture events, including natural
hybridization and anthropogenic introductions. The method developed in
this study will be useful for future population genomics studies on
medaka and on other organisms.