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Article

Genome-Wide DNA Methylation Analysis of Performance Variation in the 5000-m Speed Race of Yili Horses

1
College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China
2
Xinjiang Key Laboratory of Equine Breeding and Exercise Physiology, Urumqi 830052, China
*
Author to whom correspondence should be addressed.
Animals 2026, 16(2), 302; https://doi.org/10.3390/ani16020302
Submission received: 4 December 2025 / Revised: 6 January 2026 / Accepted: 9 January 2026 / Published: 19 January 2026
(This article belongs to the Section Animal Genetics and Genomics)

Simple Summary

Employing whole-genome bisulfite sequencing (WGBS), we generated the first single-base-resolution methylome of pre-race jugular blood in Yili horses (n = 6) contesting a 5000 m speed trial. High—versus average—performers exhibited > 10,000 CG-biassed differentially methylated regions (DMRs) concentrated in promoter and CpG-island landscapes. Hypomethylated DMRs in elite animals significantly targeted nuclear (PPARGC1A) and mitochondrial (ND1/ND2) oxidative phosphorylation genes together with neuro-muscular plasticity loci (axon guidance, glutamatergic synapse), indicating an epigenetically primed energy–neural axis conducive to explosive ATP provision and rapid motor unit recruitment. The data validate blood-derived DNA methylation signatures as non-invasive epigenetic biomarkers for early selection and tailored training of racing stock and provide novel molecular targets for precision breeding within the Yili horse population.

Abstract

Whole-genome bisulfite sequencing (WGBS) was employed in this article to map blood DNA methylation profiles at single-base resolution in Yili horses before a 5000 m speed race, with comparative analysis of epigenetic differences between the ‘elite group’ and ‘ordinary group’ across six four-year-old stallions. The overall methylation level in the elite group was generally higher than that in the ordinary groups, with a minority of regions showing hypomethylation. For instance, the promoter regions of key metabolic and neuro-related genes exhibited significant hypomethylation. The article identified over 10,000 CG differential methylation regions (DMRs), predominantly enriched in promoter and CpG island regions, anchoring 7221 differentially methylated genes (DMGs). These DMGs were significantly enriched in key biological processes including oxidative phosphorylation, protein binding, axon guidance, glutamatergic synapses, and the Hedgehog signalling pathway. Among these, six genes—ACTN3, MSTN, FOXO1, PPARGC1A, ND1, and ND2—were selected as core candidate genes closely associated with muscle strength, energy metabolism, and stress adaptation. The study confirms that the differences in athletic ability among Yili horses have a significant epigenetic basis, with DNA methylation participating in the epigenetic regulation of athletic traits by modulating the expression of genes related to energy metabolism and neuroplasticity. The constructed “promoter hypomethylated DMR panel” holds promise for translation into non-invasive blood-based epigenetic markers for early performance evaluation and targeted breeding in racehorses. This provides a theoretical basis and molecular targets for improving equine athletic phenotypes and optimising training strategies.
Keywords: Yili horse; genome-wide bisulfite sequencing; DNA methylation; differentially methylated regions; differentially methylated sites Yili horse; genome-wide bisulfite sequencing; DNA methylation; differentially methylated regions; differentially methylated sites

Share and Cite

MDPI and ACS Style

Shan, D.; Yao, X.; Ren, W.; Huang, Q.; Su, Y.; Li, Z.; Li, L.; Wang, R.; Ma, S.; Wang, J. Genome-Wide DNA Methylation Analysis of Performance Variation in the 5000-m Speed Race of Yili Horses. Animals 2026, 16, 302. https://doi.org/10.3390/ani16020302

AMA Style

Shan D, Yao X, Ren W, Huang Q, Su Y, Li Z, Li L, Wang R, Ma S, Wang J. Genome-Wide DNA Methylation Analysis of Performance Variation in the 5000-m Speed Race of Yili Horses. Animals. 2026; 16(2):302. https://doi.org/10.3390/ani16020302

Chicago/Turabian Style

Shan, Dehaxi, Xinkui Yao, Wanlu Ren, Qiuping Huang, Yi Su, Zexu Li, Luling Li, Ran Wang, Shikun Ma, and Jianwen Wang. 2026. "Genome-Wide DNA Methylation Analysis of Performance Variation in the 5000-m Speed Race of Yili Horses" Animals 16, no. 2: 302. https://doi.org/10.3390/ani16020302

APA Style

Shan, D., Yao, X., Ren, W., Huang, Q., Su, Y., Li, Z., Li, L., Wang, R., Ma, S., & Wang, J. (2026). Genome-Wide DNA Methylation Analysis of Performance Variation in the 5000-m Speed Race of Yili Horses. Animals, 16(2), 302. https://doi.org/10.3390/ani16020302

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