Background: Manual topping is a routine agronomic practice for balancing the vegetative and reproductive growth of cotton (Gossypium hirsutum) in China, but its cost-effectiveness has decreased over time. Therefore, there is an urgent need to replace manual topping with new approaches, such as biological topping. In this study, we examined the function of GhREV transcription factors (a class III homeodomain-leucine zipper family, HD-ZIP III) in regulating the development of shoot apical meristem (SAM) in cotton with the purpose of providing candidate genes for biological topping of cotton in the future. Results: We cloned four orthologous genes of AtREV in cotton, namely GhREV1, GhREV2, GhREV3, and GhREV4. All the GhREVs expressed in roots, stem, leaves, and SAM. Compared with GhREV1 and GhREV3, the expression level of GhREV2 and GhREV4 was higher in the SAM. However, only GhREV2 had transcriptional activity. GhREV2 is localized in the nucleus; and silencing it via virus-induced gene silencing (VIGS) produced an abnormal SAM. Two key genes, GhWUSA10 and GhSTM, which involved in regulating the development of plant SAM, showed about 50% reduction in their transcripts in VIGS-GhREV2 plants.Conclusion: GhREV2 positively regulates the development of cotton SAM by regulating GhWUSA10 and GhSTM potentially.