This study evaluated the association between genotypes of Leptin (LEP), Toll-like receptor 4 (TLR4), and Chemokine receptor of interleukin 8 C-X-C motif (CXCR1) genes and fertility traits in Czech Fleckvieh cattle. In LEP g.-963C>T the TT genotype heifers firstly calved 24 days earlier than CC genotype and the CT genotype cow showed a tendency for shorter days open (DO) and higher pregnancy rate. In LEP c. 357C>T we observed longer calving interval (CLI) and DO period in TT cows. Hétérozygotes in TLR4 g.-226C>G was significantly associated with shorter CLI. In CXCR1 c.777C>G we did not observe any relationship of this SNP with reproduction. Overall, the results showed that LEP and TLR4 could be effective markers for improving reproduction in Czech Fleckvieh cattle.