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.
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Overview
This study investigated DNA methylation differences in Yili horses to understand how epigenetic factors influence their performance in 5000-meter speed races.
Researchers compared the blood DNA methylation profiles of elite and ordinary horses to identify epigenetic markers associated with athletic ability.
Background and Objectives
Yili horses are a breed used in competitive racing, with some individuals displaying superior athletic performance.
The study aimed to explore epigenetic mechanisms, specifically DNA methylation, underlying performance variability.
DNA methylation involves adding methyl groups to DNA, potentially regulating gene expression without altering the genetic code.
Understanding these epigenetic changes could help identify biomarkers for breeding and training strategies.
Methods
Whole-genome bisulfite sequencing (WGBS) was used to map DNA methylation at single-base resolution.
Blood samples were taken from six four-year-old Yili stallions categorized into ‘elite’ and ‘ordinary’ groups based on race performance.
Comparisons were made to detect differences in methylation patterns, particularly in promoter regions and CpG islands.
Differentially methylated regions (DMRs) and genes (DMGs) were identified and analyzed for biological relevance.
Key Findings
The overall DNA methylation level was higher in elite horses compared to ordinary ones, indicating a general epigenetic distinction.
A subset of regions, particularly promoters of genes involved in metabolism and neurological functions, were hypomethylated in elite horses, suggesting increased gene activity in these areas.
More than 10,000 CG DMRs were found, mainly located within promoter and CpG island regions.
7221 differentially methylated genes were linked to biological processes such as:
Oxidative phosphorylation (energy production in cells)
Protein binding (cellular interactions)
Axon guidance (nerve development)
Glutamatergic synapses (neurotransmission)
Hedgehog signaling pathway (cell growth and differentiation)
Six candidate genes were highlighted for their association with physical performance:
ACTN3: muscle fiber function and strength
MSTN: regulation of muscle growth
FOXO1: stress response and metabolism
PPARGC1A: energy metabolism and mitochondrial function
ND1 and ND2: components of mitochondrial respiratory chain linked to energy production
Implications
The study demonstrated a clear epigenetic basis for differences in athletic performance among Yili horses.
DNA methylation changes appear to regulate genes critical for muscle strength, energy use, and nervous system function, all important for racing success.
The identified “promoter hypomethylated DMR panel” could be used to develop blood-based tests to evaluate horse performance potential non-invasively.
This offers promising molecular targets for selective breeding programs aiming to enhance racehorse athleticism.
It also provides a foundation for optimizing training by understanding the epigenetic regulation of key physiological traits.
Conclusion
This research bridges genomics and performance by linking epigenetic modifications to athletic traits in horses.
It pioneers use of whole-genome methylation profiling to reveal how gene regulation differences correspond to variations in speed race performance.
The findings support further exploration of epigenetic biomarkers for improving equine sports performance and personalized training regimens.
Cite This Article
APA
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 (Basel), 16(2), 302.
https://doi.org/10.3390/ani16020302
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