A functional genetic analysis in flour beetles (Tenebrionidae) reveals an antennal identity specification mechanism active during metamorphosis in Holometabola

The antenna was the first arthropod ventral appendage to evolve non-leg identity. Models of antennal evolution have been based on comparisons of antennal and leg identity specification mechanisms in Drosophila melanogaster, a species in which appendages develop from highly derived imaginal discs during the larval period. We test for conservation of the Drosophila antennal identity specification mechanism at metamorphosis in Tribolium castaneum and three other flour beetle species (Tribolium confusum, Tribolium brevicornis and Latheticus oryzae) in the family Tenebrionidae. In Drosophila, loss of function of four transcription factors-homothorax, extradenticle, Distal-less, and spineless-causes large-scale transformations of the antenna to leg identity. Distal-less and spineless function similarly during metamorphosis in T. castaneum. RNA interference (RNAi) targeting homothorax (hth) or extradenticle (exd) caused transformation of the proximal antenna to distal leg identity in flour beetles, but did not affect the identity of the distal antenna. This differs from the functional domain of these genes in early instar Drosophila, where they are required for identity specification throughout the antenna, but matches their functional domain in late instar Drosophila. The similarities between antennal identity specification at metamorphosis in flour beetles and in late larval Drosophila likely reflect the conservation of an ancestral metamorphic developmental mechanism. There were two notable differences in hth/exd loss of function phenotypes between flies and beetles. Flour beetles retained all of their primary segments in both the antenna and legs, whereas flies undergo reduction and fusion of primary segments. This difference in ground state appendage morphology casts doubt on interpretations of developmental ground states as evolutionary atavisms. Additionally, adult Tribolium eyes were transformed to elytron-like structures; we provide a developmental hypothesis for this evolutionarily surprising transformation.