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【目的】本研究旨在筛选小菜蛾Plutella xylostella应对玫烟色棒束孢Isaria fumosorosea侵染的免疫应答及其网络调控基因,以进一步探讨小菜蛾对玫烟色棒束孢的防御机制。【方法】采用第二代高通量测序技术RNA-seq,对处理后12 h的感染玫烟色棒束孢和健康(平行对照)的小菜蛾4龄幼虫转录组进行了测序,通过生物信息学分析对得到的差异基因进行了功能注释和分类,对差异基因参与的信号通路进行了分析。【结果】在感菌和健康小菜蛾幼虫转录组两个表达谱里,分别获得了12 346 987和12 315 210个clean reads,有60.93%和61.26%的reads分别能比对到参考基因库里,其中完美匹配(perfect match)的比例分别为32.15%和32.73%。共得到351个显著差异表达基因(differentially expressed unigenes,DEUs),上调表达基因有275个,下调表达基因有76个,与免疫防御反应潜在相关的基因有156个。GO(Gene Ontology)富集分析有102个DEUs分布到46个GO term里,KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway富集分析结果显示,有132个DEUs显著富集在13个代谢通路(pathway)里。【结论】这些差异表达基因中,大部分编码潜在的与免疫识别及调控相关的基因,集中在能量代谢、疾病反应和防御反应等相关通路。研究结果为挖掘与玫烟色棒束孢侵染相关的小菜蛾免疫应答候选基因提供了重要数据库,也为阐明小菜蛾对玫烟色棒束孢的免疫机制奠定理论基础。
【Objective】 The aim of this study was to screen for the immune response and its network regulatory genes of Plutella xylostella against Isaria fumosorosea infection in Plutella xylostella to further explore the defense mechanism of Plutella xylostella against Plutella xylostella against Plutella xylostella. 【Method】 The fourth generation larvae of the diamondback moth, Plutella xylostella, infected with C. aeruginosa and the healthy (parallel control) 12-h post-treatment were sequenced using the second-generation high-throughput sequencing technique RNA-seq. The functional analysis and classification of the differentially expressed genes were analyzed, and the signal pathways involved in the differential gene were analyzed. 【Result】 The results showed that 12 346 987 and 12 315 210 clean reads were obtained in the two expression profiles of the susceptible and healthy Plutella xylostella larvae, respectively, and 60.93% and 61.26% of the reads were aligned to the reference gene pool , Of which the proportions of perfect match were 32.15% and 32.73% respectively. A total of 351 differentially expressed unigenes (DEUs) were obtained, including 275 up-regulated genes, down-regulated genes from 76, and 156 genes potentially related to immune defense response. Enrichment analysis of GO (Gene Ontology) 102 DEUs were distributed in 46 GO terms, enrichment analysis of KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway showed that 132 DEUs were significantly enriched in 13 metabolic pathways )in. 【Conclusion】 Most of these differentially expressed genes encode potential genes involved in immune recognition and regulation, focusing on energy metabolism, disease response and defense response. The results provide an important database for mining the candidate genes of immune response to diamondback moth invader Plutella xylostella and lay a theoretical foundation for elucidating the immune mechanism of Plutella xylostella to C. litura.