MicroRNAs (miRNAs) are a group of short non-coding RNAs involved in the inhibition of protein translation or in mRNA degradation. Although the regulatory roles of miRNAs in various biological processes have been investigated, there is as yet an absence of studies about the regulatory roles of miRNAs involved in the metabolism of plant allelochemicals in insects. We constructed five small RNA libraries from apterous Aphis gossypii adults that had fed on an artificial diet containing various allelochemicals. Using Illumina sequencing, a total of 73.27 million clean reads was obtained, and 292 miRNAs were identified from A. gossypii. Comparative analysis of read counts indicated that both conserved and novel miRNAs were differently expressed among the five libraries, and the differential expression was validated via qRT-PCR. We found that the transcript levels of several miRNAs were increased or decreased in all of the allelochemical treatment libraries compared to the control. The putative target genes of the miRNAs were predicted using in silico tools, and the target genes of several miRNAs were presumed to be involved in the metabolism of xenobiotic compounds. Furthermore, the target prediction results were confirmed using dual luciferase reporter assay, and Ago-miR-656a-3p was demonstrated to regulate the expression of CYP6J1 post-transcriptionally through binding to the 3′ UTR of CYP6J1. Our research results indicate that miRNAs may be involved in the metabolism of plant allelochemicals in A. gossypii, and these results also represent an important new small RNA genomics resource for further studies on this topic.

中文翻译：

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鉴定棉蚜中的microRNA及其对植物化感物质胁迫的响应（半翅目：蚜科）。

MicroRNA（miRNA）是一组短的非编码RNA，参与抑制蛋白质翻译或mRNA降解。尽管已经研究了miRNA在各种生物过程中的调控作用，但目前尚无关于miRNA参与昆虫植物化感物质代谢的调控作用的研究。我们从无性蚜虫成虫中构建了五个小RNA文库，这些成虫以人工饮食为食，其中含有各种化感物质。使用Illumina测序，总共获得了7327万条纯净读数，并从棉球菌中鉴定出292个miRNA。读取计数的比较分析表明，在五个文库中保守和新型miRNA的表达均不同，并且通过qRT-PCR验证了差异表达。我们发现，与对照相比，在所有化感处理库中，几种miRNA的转录水平均升高或降低。使用计算机模拟工具预测了miRNA的推定靶基因，并推测了几种miRNA的靶基因参与了异源化合物的代谢。此外，使用双荧光素酶报告基因测定法证实了目标预测结果，并且证明了Ago-miR-656a-3p通过与CYP6J1的3'UTR结合来调节转录后CYP6J1的表达。我们的研究结果表明，miRNA可能参与了棉铃虫中植物化感物质的代谢，这些结果也代表了一个重要的新小RNA基因组学资源，可供对该主题进行进一步研究。