[Aim]  This study aims to investigate the effect of royal jelly (RJ) on honeybee learning and memory to gain a greater understanding of the underlying mechanisms of honeybee memory. [Methods]  After emergence, queens and workers were raised in cages and fed 0% RJ, 10% RJ, and 40% RJ dissolved in 30% sucrose. Five days post-emergence we tested their learning, memory, and odor discrimination using the appetitive olfactory related proboscis extension reflex (PER) paradigm. Given the critical role of Dnmt3 gene in honeybee acquisition and extinction learning and memory formation, we tested the Dnmt3 gene expression pattern by real-time quantitative PCR in the brain of queens and workers fed RJ. [Results]  Results of the PER test indicate that learning, memory, and odor discrimination was significantly improved (worker: F_2,1805=35.29, P<0.000 1; queen: F_2,909=8.69, P=0.000 2) in both castes that were fed 40% RJ. However, odor discrimination was higher in workers compared to queen bees. Gene testing showed that Dnmt3 gene expression was upregulated in worker bees but downregulated in queen bees following a diet of 40% RJ. Here we provide evidence that a high concentration of RJ can improve honeybee cognition and influence the expression pattern of the Dnmt3 gene. [Conclusion]  These findings offer valuable insights into the role of RJ in facilitating honeybee learning and memory.