Primer sequences used in this study.

To understand the mechanisms underlying social evolution and caste development in social insects, caste-specific organs and genes should be investigated. In the rhinotermitid termite, Reticulitermes speratus, the lipocalin gene RS008881, which encodes a protein transporter, is expressed in the ovarian accessory glands of primary queens. To obtain additional data on its expression and product localization, we conducted real-time quantitative polymerase chain reaction and protein assays using a peptide antibody. Gene expression analysis of the castes revealed that RS008881 was highly expressed in female primary and secondary reproductives. Further analysis of its expression during reproductive caste differentiation showed that its expression levels increased prior to molting into reproductive individuals, even during the winged imago (alates) stage. Western blotting and fluorescent immunohistochemical staining revealed that the RS008881 product was localized in the ovary as well as the eggshells produced by female reproductives. RS008881 may play a significant role in the reproductive biology of R. speratus; protein localization in both the ovary and eggshell suggests multiple functions related to embryo protection and potential pheromone interactions.