The Bicyclus lineage of satyrid butterflies exhibits male-specific traits, the scent organ 9 complex, used for chemical communication during courtship. This complex consists of tightly 10 packed brush-like scales (hair-pencils) that rub against scent patches to disperse pheromones, but 11 the evolution and molecular basis of the organ's male-limited development remains unknown. 12 Here, we examine the evolution of the number and location of the scent patches and hair-pencils 13 within 53 species of Bicyclus butterflies, and the involvement of the sex determinant gene 14 doublesex (dsx) in scent organ development in Bicyclus anynana using CRISPR/Cas9. We show 15 that scent patches and hair-pencils arose via multiple, independent gains, in a correlated manner. 16 Further, an initially non-sex-specific Dsx protein expression pattern in developing wing discs 17 becomes male-specific and spatially refined to areas that develop the scent organ complex over 18 the course of development. Functional perturbations of dsx show that this gene is required for 19 male patch development whereas hair-pencils can develop in both sexes without Dsx input. Dsx 20 in females is, instead, required to repress hair-pencils. These findings suggest that the patches 21 and hair-pencils evolve as correlated composite organs that are sex-limited via the spatial 22 regulation of dsx. Divergence in the function of dsx isoforms occurs in both sexes, where the 23 male isoform promotes patch development in males and the female isoform represses hair-pencil 24 development in females, both leading to the development of male-limited traits. Furthermore, 25 evolution in number and location of patches, but not of hair-pencils, appears to be regulated by 26 spatial regulation of dsx. 27 28 65On the other hand, many species in the tribe Satyrini possess scent organs consisting of hair-66 pencils (Fig 1 A,B) and scent patches (Fig 2A), both on the wing, that brush against each other 67 dispersing chemicals in the process (Nieberding et al. 2008;Brattström et al. 2015Brattström et al. , 2016 Aduse-68 Poku et al. 2017). It is to be noted however, that glandular, secretory cells underly some of the 69 130 (hair-pencil 3 without patch 3 in B. jefferyi, B. moyses and B. dorothea) and vice versa (patch 13 131 in B. hyperanthus, B. scaithis and B. elishiae). 132 133 Patterns of androconia diversity within Bicyclus occurs primarily via multiple trait gain, 134 and trait loss 135In order to understand the origins of this diversity within Bicyclus, we first constructed a 136 phylogenetic tree of the species of interest using a Bayesian framework and then reconstructed 137 the evolutionary history of the different hair-pencils and patches on the sampled trees. Our 138