Novel seminal fluid proteins in the seed beetle Callosobruchus maculatus identified by a proteomic and transcriptomic approach
Novel seminal fluid proteins in the seed beetle Callosobruchus maculatus identified by a proteomic and transcriptomic approach
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DOI：
10.1111/imb.12271
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发表时间：
2017-02-01
影响因子：
2.6
通讯作者：
Arnqvist, G.
中科院分区：
文献类型：
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作者：
Bayram, H.;Sayadi, A.;Arnqvist, G.
种子甲虫Callosobruchus maculatus是一种重要的农业害虫，也是一种两性冲突模式。男性拥有生殖器棘，可以增加精液蛋白（SFP）进入女性身体的转移。由于SFPs改变了雌性的行为和生理，它们可能会调节该物种的生殖和性冲突。在这里，我们使用蛋白质组学与从头转录组相结合来鉴定SFPs。先前的2D-十二烷基硫酸钠聚丙烯酰胺凝胶电泳分析确定了可能在交配时转移到雌性的雄性附属腺蛋白点。这些点的蛋白质组学分析确定了98种蛋白质，其中大部分也存在于从女性收集的精液中。标准注释工作流程揭示了SFP的常见官能团，包括蛋白酶和代谢蛋白。转录组学分析发现84个转录本在性别之间差异表达。值得注意的是，编码15种蛋白质的基因在雄性果蝇中高度表达，而在雌性果蝇中的表达可以忽略不计。这些序列中的大多数对应于“未知”的蛋白质（15个中的9个），并可能代表快速发展的新的种子甲虫的SFP。我们的综合分析突出了44种蛋白质，有强有力的证据表明它们是SFP。这些结果可以为进一步的研究提供信息，以更好地了解种子甲虫性冲突的分子机制。
The seed beetle Callosobruchus maculatus is a significant agricultural pest and increasingly studied model of sexual conflict. Males possess genital spines that increase the transfer of seminal fluid proteins (SFPs) into the female body. As SFPs alter female behaviour and physiology, they are likely to modulate reproduction and sexual conflict in this species. Here, we identified SFPs using proteomics combined with a de novo transcriptome. A prior 2D-sodium dodecyl sulphate polyacrylamide gel electrophoresis analysis identified male accessory gland protein spots that were probably transferred to the female at mating. Proteomic analysis of these spots identified 98 proteins, a majority of which were also present within ejaculates collected from females. Standard annotation workflows revealed common functional groups for SFPs, including proteases and metabolic proteins. Transcriptomic analysis found 84 transcripts differentially expressed between the sexes. Notably, genes encoding 15 proteins were highly expressed in male abdomens and only negligibly expressed within females. Most of these sequences corresponded to 'unknown' proteins (nine of 15) and may represent rapidly evolving SFPs novel to seed beetles. Our combined analyses highlight 44 proteins for which there is strong evidence that they are SFPs. These results can inform further investigation, to better understand the molecular mechanisms of sexual conflict in seed beetles.