Institute of Entomology and Molecular Biology, College of Life Sciences, Chongqing Normal University, Chongqing 401331, China.
J Insect Physiol. 2012 Nov;58(11):1444-9. doi: 10.1016/j.jinsphys.2012.08.010. Epub 2012 Sep 15.
To gain a better understanding of the molecular mechanisms regulating pupal diapause of the onion maggot Delia antiqua, PCR-based suppressive subtractive hybridization was performed to identify genes involved in summer and/or winter diapause. A total of 209 unique sequences were obtained including 89 in forward library for winter diapausing pupae and 120 in the reverse library for summer diapausing pupae. 76.4% (68/89) and 68.3% (82/120) unique sequences had significant hits to non-redundant proteins database. Gene functional annotation showed these non-redundant sequences are involved in stress response and innate immunity, metabolism and energy, information processing and regulation, binding, food storage, morphogenesis and development, cell skeleton and cycle, protein synthesis and folding. Approximately 28.2% (59/209) transcripts showed no significant similarity to any other sequence in the public databases, probably representing unique genes of the onion maggot. Semi-quantitative RT-PCR revealed that the relative expression levels of 18 genes were comparable between summer and winter diapause. This study elucidates the temporal expression of diapause-related genes in onion maggot, also provides new insights into the differences in the physiological changes in summer and winter pupae. Functional characterization of some candidate genes will further enhance the understanding of the generating, maintaining, and breaking mechanism of diapause.
为了更好地了解调控洋葱地种蝇蛹滞育的分子机制,我们进行了基于 PCR 的抑制性消减杂交,以鉴定参与夏眠和/或冬眠的基因。总共获得了 209 个独特的序列,包括冬季滞育蛹正向文库中的 89 个和夏季滞育蛹反向文库中的 120 个。76.4%(89/117)和 68.3%(120/176)的独特序列与非冗余蛋白质数据库有显著的匹配。基因功能注释表明,这些非冗余序列参与应激反应和先天免疫、代谢和能量、信息处理和调节、结合、食物储存、形态发生和发育、细胞骨架和周期、蛋白质合成和折叠。大约 28.2%(59/209)的转录本与公共数据库中的任何其他序列没有显著相似性,可能代表洋葱地种蝇的独特基因。半定量 RT-PCR 显示,18 个基因在夏季和冬季滞育之间的相对表达水平相当。本研究阐明了洋葱地种蝇滞育相关基因的时空表达,也为夏眠和冬眠蛹生理变化的差异提供了新的见解。一些候选基因的功能表征将进一步增强对滞育产生、维持和打破机制的理解。