Endocrine system in supernumerary molting of the flour beetle, Triboliwn freemani, under crowded conditions

In the flour beetle, Tribolium freemani, a crowded environment in the last larval instar delays the development into a pupa, but the beetle continues to engage in larval-larval molting, which is an adaptive response to avoid being the victim of cannibalism as an immobile pupa. To understand the endocrine mechanism involved in this developmental process, we investigated the components of the juvenile hormone and ecdysone signaling systems. We examined whether elevated juvenile hormone levels in the crowded condition is the sole causal factor for the supernumerary molting. RNA interference (RNAi) of the JH acid methyltransferase (TfMT3) for lowering juvenile hormone titer in the crowded condition could not rescue pupation and instead resulted in lethality with developmental arrest at the prepupal stage. Kruppel-homolog 1 (7fKr-h1), the immediate downstream JH signal, was highly upregulated even in the RNAi of TfMT3 in a crowded condition. RNAi of TfKr-h1 resulted in a phenocopy of the lethal TfMT3 RNAi in a crowded condition. In addition, RNAi of TfM7'3 in a crowded condition resulted in lack of the major ecdysone peak in the prepupal stage. We conclude that while a crowded condition induces supernumerary molts by elevating juvenile hormone levels, it can also inhibit metamorphosis by disrupting additional endocrine processes. The current study suggests that crowded conditions affect multiple independent factors in the endocrine and the downstream signaling systems.