Dissecting the Isoform-Specific Roles of FTZ-F1 in the Larval-Larval and Larval-Pupal Ecdyses in Henosepilachna vigintioctopunctata

Simple Summary Fushi Tarazu Factor-1 (FTZ-F1) plays a crucial regulatory role in molting in insects. It is hypothesized that, by alternative transcription start and splicing, the FTZ-F1 gene generates two isomers (alpha- and beta FTZ-F1) that exert isoform-specific roles in non-Drosophilid insects. In the present paper, we first unveiled that the same post-transcriptional processing in FTZ-F1 occurred in coleopterans, lepidopterans, dipterans and hymenopterans. We then found that alpha FTZ-F1 and beta FTZ-F1 were actively transcribed throughout the development, from embryo to adult, in Henosepilachna vigintioctopunctata. Moreover, by RNA interference, we confirmed that both FTZ-F1 isoforms act as regulators in larval-larval molting and beta FTZ-F1 is involved in the regulation of the larval-pupal transition. Fushi Tarazu Factor 1 (FTZ-F1), a member of the nuclear receptor superfamily, is the downstream factor of 20-hydroxyecdysone signaling. In Drosophila melanogaster, alternative transcription start and splicing in the FTZ-F1 gene generate alpha FTZ-F1 and beta FTZ-F1 isoforms, which are vital for pair-rule segmentation in early embryogenesis and post-embryonic development, respectively. However, whether the same mRNA isoforms are present and exert the conservative roles remains to be clarified in other insects. In the present paper, we first mined the genomic data of representative insect species and unveiled that the same post-transcriptional processing in FTZ-F1 occurred in coleopterans, lepidopterans, dipterans and hymenopterans. Our expression data in Henosepilachna vigintioctopunctata, a serious polyphagous defoliator damaging a wide range of crops in Solanaceae and Cucurbitaceae, showed that both alpha FTZ-F1 and beta FTZ-F1 were actively transcribed throughout the development, from embryo to adult. The RNA interference-aided knockdown of both isoforms completely arrested larval ecdysis from the third to the fourth instar, in contrast to the depletion of either isoform. In contrast, silencing beta FTZ-F1, rather than alpha FTZ-F1, severely impaired the larval-pupal transformation. We accordingly propose that both FTZ-F1 isoforms are essential but mutually interchangeable for larval-larval molting, while beta FTZ-F1 is necessary for the larval-pupal transition and sufficient to exert the role of both FTZ-F1s during larval-pupal metamorphosis in H. vigintioctopunctata.

Automated summary

FTZ-F1 plays a crucial regulatory role in molting in insects, and the functions of two isoforms were confirmed through RNA interference.