基于转录组和基因表达谱的葱蝇分泌组挖掘及其在滞育发育中的可能作用分析（英文）

Mining the secretome of Delia antiqua(Diptera:Anthomyiidae)based on the transcriptomic and gene expression data and analysis of its potential roles in diapause development

【目的】昆虫滞育是一种重要的季节性适应生存的环境生理现象,但是调控葱蝇Delia antiqua夏滞育和冬滞育的分子机理却知之甚少。本研究旨在鉴定与葱蝇蛹滞育相关的候选基因。【方法】利用RNA-seq和数字基因表达谱数据,通过生物信息学的方法预测分析葱蝇滞育相关的分泌蛋白,并进行了氨基酸序列相似性、基本理化性质分析;从表达谱中选取12个差异表达基因,利用qRT-PCR验证了它们在非滞育蛹以及夏滞育和冬滞育蛹的不同发育时期的表达特点。【结果】共鉴定出38个在葱蝇夏滞育蛹和冬滞育蛹之间差异表达的可能的分泌蛋白。同源基因注释表明,这些基因可能在气味分子结合、几丁质和脂类代谢、免疫及发育等方面发挥作用,其中脂类和几丁质代谢过程的调节似乎在滞育发育过程起到重要作用。qRT-PCR分析结果表明,12个被选择基因的表达谱与RNA-seq结果相吻合(R=0.862,P=0.001)。【结论】本研究为基于高通量测序的分泌组挖掘及滞育相关基因的鉴定提供了强有力地分析策略。

【Aim】 Insect diapause is a critical eco-physiological adaptation for seasonal survival, but little is known about the molecular mechanisms of summer and winter diapause of Delia antiqua. Here, our main goal is to identify candidate genes linked to pupal diapause of D. antiqua. 【Methods】 A straightforward in silico approach was performed to predict the secreted diapause-related proteins based on the RNA-seq and digital gene expression datasets. Amino acid sequence similarity and basic physiochemical characteristics analysis were conducted. Gene expression levels of 12 selected genes from RNA-seq data were further validated in non-diapause pupae, and different developmental stages of summer and winter diapause pupae by qRT-PCR. 【Results】 A total of 38 putative secreted proteins were differentially regulated between the summer and winter diapause pupae of D. antiqua. Gene Ontology analysis revealed differentially expressed genes to be associated with odorant binding, chitin and lipid metabolic processes, innate immunity and development, etc. Among them, regulation of lipid mobilization and chitin-related metabolism was found to be important during diapause development. qRT-PCR analyses showed that the expression profiles of the 12 selected genes were in good agreement with results from RNA-seq （R=0.862, P=0.001）. 【Conclusion】 This study establishes a robust pipeline for the discovery of diapause-related genes by secretome mining based on the high-throughput sequencing.