Department of Biology, Georgetown University, Reiss 406, Washington, DC 20057, USA.
J Insect Physiol. 2010 Sep;56(9):1147-54. doi: 10.1016/j.jinsphys.2010.03.008. Epub 2010 Mar 21.
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.
许多温带昆虫通过进入光周期休眠来应对冬季的不利条件，但这种关键适应的分子基础还不是很清楚。在白纹伊蚊的温带种群中，暴露在短日照长度下会导致成年雌性产卵休眠。对来自温带白纹伊蚊种群的成熟（V 期）卵母细胞的 RNA 进行抑制性消减杂交（SSH）和定量逆转录 PCR（qRT-PCR）。从 SSH 文库中测序了 438 个插入片段，鉴定了 324 个独特（非冗余）序列。使用包括经历光周期休眠的温带种群和不经历光周期休眠的热带种群的重复实验设计，对 53 个转录物进行了 QRT-PCR 实验。在多个温带种群中，短日照与长日照相比，上皮膜蛋白转录物的丰度更高，但在多个热带种群中并非如此。该基因可能通过增加抗干燥能力或能量储备在休眠程序中发挥作用。与 LD 光周期相比，SD 光周期对磷酸乙醇胺结合蛋白转录物的表达在温带和热带种群之间存在差异，但似乎与休眠无关。进行了快速 cDNA 末端扩增（RACE）以确定这两个转录物的全长 cDNA 序列。
