A transcriptional element of the diapause program in the Asian tiger mosquito, Aedes albopictus, identified by suppressive subtractive hybridization

Many temperate insects cope with the unfavorable conditions of winter by entering a photoperiodic diapause, but the molecular basis of this crucial adaptation is not well understood. In temperate populations of Aedes albopictus, exposure to short-day lengths causes the adult female to oviposit diapausing eggs. Suppressive subtractive hybridization (SSH) and quantitative reverse transcription PCR (qRT-PCR) were performed on RNA isolated from mature (stage V) oocytes of a temperate population of Ae. albopictus. A total of 438 inserts were sequenced from the SSH library and 324 unique (non-redundant) sequences were identified. QRT-PCR experiments were performed for 53 transcripts using a novel experimental design that included replicate temperate populations that do undergo photoperiodic diapause and replicate tropical populations that do not undergo a photoperiodic diapause. There was greater abundance of an epithelial membrane protein transcript under short-day versus long-day photoperiods in multiple temperate, but not tropical, populations. This gene may function during the diapause program by increasing desiccation resistance or energy reserves. The expression of a phosphatidylethanolamine-binding protein transcript in response to SD versus LD photoperiod differed between temperate and tropical populations but does not appear to be causally involved in diapause. Rapid Amplification of cDNA Ends (RACE) was performed to determine the entire cDNA sequence of both transcripts.