Histone modifying genes are involved in the molting period during soldier differentiation in Zootermopsis nevadensis

Caste differentiation in eusocial insects is an outstanding example of phenotypic plasticity. Recent studies indicate that epigenetic regulation, including DNA methylation and histone modification, play a role in the morphological and behavioral polyphenism observed in the caste differentiation of hymenopteran insects. The role of epigenetic regulation in termite caste differentiation, however, is still obscure. In this study, we performed a functional analysis of epigenetic-related genes during soldier differentiation in Zootermopsis nevadensis, for which the entire genome sequence is available. In an incipient colony of this species, the oldest 3rd instar larva (No. 1 larva) always differentiates into a presoldier (intermediate stage of soldier), and the next-oldest 3rd instar larva (No. 2 larva) molts into a 4th instar (which functions as a worker). First, we detected seven epigenetic-related genes with significantly increased expression levels in the soldier-destined No. 1 larvae using RNA-seq data. Second, RNA interference (RNAi) of these seven genes was performed in the No. 1 larvae. RNAi of three histone modifying genes extended the presoldier molting period. Furthermore, these RNAi treatments reduced the expression levels of genes involved in juvenile hormone (JH) synthesis, binding and signaling. These results indicate that epigenetic-related genes do not directly affect termite soldier differentiation; nonetheless, some histone modifying genes have an effect on molting periods, possibly due to the regulation of JH action during soldier differentiation.