Vitamin D deficiency can negatively impact the health and training efficiency of athletes. The rs10741657(G) allele in CYP2R1 gene has been associated with lower vitamin D status, potentially contributing to vitamin D deficiency observed across various populations, including Indians. This study aimed to investigate whether the CYP2R1 (rs10741657) polymorphism is contributing to the vitamin D status among Indian athletes. Blood samples were collected from 92 Indian elite and sub-elite athletes participating in weightlifting (29), badminton (32), athletics (14), and rowing (17) for laboratory analysis. PCR-RFLP of the CYP2R1 (rs10741657) SNP was performed, and vitamin D levels were measured. Statistical analysis was done to assess the association between genotype and vitamin D status. Vitamin D levels were comparable between genders but differed across sports, with indoor athletes, especially badminton players having higher levels. The AA genotype was associated with higher vitamin D levels compared to the GG and AG genotypes. Vitamin D deficiency is highly prevalent among the athletes studied. This deficiency may be due to the higher prevalence of the alternate allele (G) of CYP2R1 polymorphism, suggesting that genetic predisposition plays a key role in determining the vitamin D status of athletes.