It is a fundamental challenge to discover the association of genetic traits with phenotypic traits. In this study, we aimed to identify possible genetic traits related to horse temperament. Based on previous findings, we selected 71 candidate genes related to temperamental trait and examined them in the human and horse reference genomes (hg38 and equCab2, respectively). We found 16 orthologous genes and, by comparing with the human reference genome, 17 homologous genes in the horse reference genome. We designed probes specific for the 33 horse genes. Using the probes, we built sequencing libraries of the genomic DNA samples from eight aggressive and eight docile horses, and sequenced the constructed libraries using the Illumina Hiseq2500 platform. Through the analysis of the targeted exome sequences, we identified single nucleotide polymorphisms (SNPs) in the genes. SNPs could be served as genetic markers to evaluate aggressive or docile levels of horses. To examine whether any genetic variants are associated with horse temperament, we performed genome-wide association study (GWAS) using the SNP data. GWAS analysis identified ten variants (p-value <0.05) which could be related to horse temperament. We validated the variants using Sanger sequencing. The most significant variants were found in MAOA (c.1164+41T>C) and AR (c.1047+27G>T) genes with 8.09 × 10−4 p-value. We suggest that the variants might be used to assess horse temperament and to determine superior horses for riding or racing.
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This work was supported by Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through Agri-Bio industry Technology Development Program, funded by Ministry of Agriculture, Food and Rural Affairs (MAFRA)(316026).
Conflict of interest
Soyoung song declares that she has no conflict of interest. Dong-Yep Oh declares that he has no conflict of interest. Gil-Jae Cho declares that he has no conflict of interest. Dong Hee Kim declares that she has no conflict of interest. Yong-Soo Park declares that he has no conflict of interest. Kyuong Han declares that he has no conflict of interest.
All research protocols and animal experiments in this study were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) in Gyeongsangbuk-do, Republic of Korea (Gyeongbuk IACUC-81).
The original version of this article was revised: The error in the author name has been corrected.
A correction to this article is available online at https://doi.org/10.1007/s13258-017-0627-3.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Supplementary Figure 1—Sanger sequencing validation of the SNP in NTSR1, MAOB, NLN, and DRD3 gene. The SNPs in NTSR1, MAOB, NLN, and DRD3 gene were validated by Sanger sequencing in 8 aggressive and 8 docile horses. Samples with homozygous variation were colored in dark blue box and those with heterozygous variation were colored in light blue box. (PPTX 405 KB)
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Song, S., Oh, DY., Cho, GJ. et al. Targeted next-generation sequencing for identifying genes related to horse temperament. Genes Genom 39, 1325–1333 (2017). https://doi.org/10.1007/s13258-017-0597-5
- Genetic marker
- GWAS analysis
- Horse temperament
- Targeted exome sequencing