microRNAs(miRNAs)通过调节转录后基因表达在重要的生物过程中发挥关键作用,并在发育过程中表现出差异表达模式。免疫反应,强调挑战。小菜蛾对全世界的农作物造成重大的经济损失。尽管在理解这种害虫的分子生物学方面取得了重大进展,关于miRNA在调节关键免疫相关基因中的作用的知识仍然有限.在这项研究中,我们利用小菜蛾感染绿僵菌的全转录组重新测序数据,鉴定靶向酚氧化酶原活化蛋白酶1(PAP1)基因的特异性miRNA,并在黑化过程中调节酚氧化酶(PO)级联反应.七个miRNA(pxy-miR-375-5p,pxy-miR-4448-3p,pxy-miR-279a-3p,pxy-miR-3286-3p,pxy-miR-965-5p,pxy-miR-8799-3p,和pxy-miR-14b-5p)进行筛选。荧光素酶报告基因测定证实,pxy-miR-279a-3p与开放阅读框(ORF)结合,pxy-miR-965-5p与PAP1的3'非翻译区(3'UTR)结合。我们的实验表明,pxy-miR-965-5p模拟物显著降低了小菜蛾幼虫的PAP1表达,抑制PO活性,和增加幼虫死亡率。相反,注射pxy-miR-965-5p抑制剂可增加PAP1表达和PO活性,同时降低幼虫死亡率.此外,我们鉴定了可能与pxy-miR-965-5p相互作用的4种LncRNAs(MSTRG.32910.1,MSTRG.7100.1,MSTRG.6802.1和MSTRG.22113.1).使用反义寡核苷酸(ASO)的干扰测定显示,沉默MSCRG.710.1和MSCRG.22113.1增加了pxy-miR-965-5p的表达。这些发现揭示了pxy-miR-965-5p在木雀鱼感染的免疫应答中的潜在作用,并为针对该害虫免疫系统的生物防治策略提供了理论依据。
MicroRNAs (miRNAs) play a pivotal role in important biological processes by regulating post-transcriptional gene expression and exhibit differential expression patterns during development, immune responses, and stress challenges. The diamondback moth causes significant economic damage to crops worldwide. Despite substantial advancements in understanding the molecular biology of this pest, our knowledge regarding the role of miRNAs in regulating key immunity-related genes remains limited. In this study, we leveraged whole transcriptome resequencing data from Plutella xylostella infected with Metarhizium anisopliae to identify specific miRNAs targeting the prophenoloxidase-activating protease1 (PAP1) gene and regulate phenoloxidase (PO) cascade during melanization. Seven miRNAs (pxy-miR-375-5p, pxy-miR-4448-3p, pxy-miR-279a-3p, pxy-miR-3286-3p, pxy-miR-965-5p, pxy-miR-8799-3p, and pxy-miR-14b-5p) were screened. Luciferase reporter assays confirmed that pxy-miR-279a-3p binds to the open reading frame (ORF) and pxy-miR-965-5p to the 3\' untranslated region (3\' UTR) of PAP1. Our experiments demonstrated that a pxy-miR-965-5p mimic significantly reduced PAP1 expression in P. xylostella larvae, suppressed PO activity, and increased larval mortality rate. Conversely, the injection of pxy-miR-965-5p inhibitor could increase PAP1 expression and PO activity while decreasing larval mortality rate. Furthermore, we identified four LncRNAs (MSTRG.32910.1, MSTRG.7100.1, MSTRG.6802.1, and MSTRG.22113.1) that potentially interact with pxy-miR-965-5p. Interference assays using antisense oligonucleotides (ASOs) revealed that silencing MSTRG.7100.1 and MSTRG.22113.1 increased the expression of pxy-miR-965-5p. These findings shed light on the potential role of pxy-miR-965-5p in the immune response of P. xylostella to M. anisopliae infection and provide a theoretical basis for biological control strategies targeting the immune system of this pest.