柑橘大实蝇蛹期滞育的分子机理研究

柑橘大实蝇是一种危害严重的柑橘害虫，近年来在我国多地暴发成灾，造成大量经济损失，因此受到人们广泛重视。该虫于蛹期出现专性滞育，对协调其与环境的关系具有重要的作用。研究滞育机理不仅有助于了解其如何协调与环境之间的相互关系，还能在此基础上人为解除或调控滞育，帮助解决其实验室内饲养技术的难题，也能为其综合防治提供新思路。然而，目前尚未对滞育的分子机理展开相关研究。因此，本项首先采用长期监测呼吸产生二氧化碳浓度的方法，计算拟合整个蛹期的代谢速率曲线，从而判断滞育的发生与结束。随后，在其基础上比较分析滞育准备期、滞育早、中、晚期以及滞育结束后发育期5个时间点上样品的数字基因表达谱与代谢谱差异，找出各时间点上具有特异性或显著差异的基因表达与代谢产物水平，随后利用RNAi技术对相关重要基因进行沉默等手段验证基因编码蛋白的功能。最后综合分析各研究手段的结果，达到揭示柑橘大实蝇滞育机理的目的。

Chinese citrus fly, Bactrocera (Tetradacus) minax (Enderlein), is a devastating citrus pest. The outbreaks of it in China have caused large economic losses, thus arousing increasing attentions. The obligatory diapause occurring in pupal stage plays an important role in coordinating the relationships between the pest and the environment. Therefore, study on the mechanisms underlying the diapause is conducive to understand how the diapause coordinates pest-environment relationships. Also, it contributes to artificial termination and regulation of diapause, thereby solving the problems in rearing of this pest in the laboratory and providing new thoughts in pest management. So far,the molecular mechanisms underlying diapause, however, remains unkown. So, this study will start by long-term monitoring the concentrations of carbon dioxide produced by respiration in order to fit the metabolic rate curve of whole pupal stage, based on which the entry and termination of diapause can be determined. Then, samples at pre-, early, middle, late, and post-diapause stages will be compared in terms of digital gene expression and metabolic profiling to find out the specific or significantly different gene expressions and metabolite levels. Several most important genes will subsequently be screened to investigate the function of their encoding protein in vivo through RNAi technique. At last, the results of all these approaches will be comprehensively analyzed to reveal the mechanisms underlying the pupal diapause of Chinese citrus fly.