Anther culture is a promising technique used in rice (Oryza sativa L.) breeding, although the genetic mechanisms associated with anther culturability remain elusive. In the process of in vitro anther culture, favorable alleles for anther culture are preferentially transmitted to doubled haploid (DH) populations, resulting in biased segregation of DH populations. Therefore, the segregation distortion (SD) method is an ideal approach for mapping the genetic loci controlling anther culturability. Performing a SD analysis using a genetic map consisting of 666 single nucleotide polymorphism markers, we identified five potential loci (logarithm of odds [LOD] > 3.5) that may be associated with anther culturability, including SDL2 and SDL3, which are overrepresented in japonica alleles, and SDL7, SDL9, and SDL11, which are overrepresented in indica alleles. In addition, nine pairs of epistatic interactions (EPIs) that contribute significantly (p < 0.05) to SD in the DH population were identified. Among them, the interaction between SDL2 and SDL3 is particularly noteworthy. These findings suggest that anther culturability in rice is governed by complex genetic mechanisms involving multiple nuclear genes and EPIs. This study provides insights into the genetic control underlying anther culturability in rice and lays the foundation for future research aimed at identifying causal genes associated with anther culturability. Core Ideas: A segregation distortion loci (SDLs) analysis was conducted on a doubled haploid (DH) population comprising 154 lines.Five SDLs (logarithm of odds [LOD] > 3.5) that may be associated with anther culturability were identified.Nine pairs of epistatic interactions that contribute significantly to SD in the DH population were identified.Rice culturability may be governed by complex genetic mechanisms with multiple genes and epistatic interactions. [ABSTRACT FROM AUTHOR]