Table_3_Comprehensive Identification and Characterization of Long Non-coding RNAs Associated With Rice Black-Streaked Dwarf Virus Infection in Laodelphax striatellus (Fallén) Midgut.XLSX

Long non-coding RNAs (lncRNAs) are involved in a variety of biological functions through transcriptional and post-transcriptional regulation. However, little is known about their functions in the process of insect mediated virus transmission. In the present study, we analyzed using RNA-Seq, the lncRNAs that were differentially expressed in response to Rice black-streaked dwarf virus (RBSDV) infection in Laodelphax striatellus (Fallén) midgut. A total of 13,927 lncRNAs were identified and over 69% were assigned to intergenic regions. Among them, 176 lncRNAs were differentially expressed and predicted to target 168 trans-regulatory genes. Ten differentially expressed lncRNAs were selected and their expression changes were validated by RT-qPCR. KEGG analysis showed that these target genes were enriched in the essential biological process, such as Purine metabolism, Valine, leucine and isoleucine degradation, and Fatty acid elongation. The expression levels of the differentially expressed lncRNAs and the predicted target genes that were significantly enriched in the Human papillomavirus infection pathway were analyzed by RT-qPCR. The results showed that several lncRNAs were co-expressed with their target genes. One of the lncRNAs called MSTRG15394 and its target gene, encoding a secreted protease inhibitor (PI), were up-regulated at the transcriptional level after RBSDV infection. Knockdown of MSTRG15394 could down-regulate the PI expression at mRNA level. Inhibition of either MSTRG15394 or PI expression by RNA interference promoted RBSDV accumulation in L. striatellus midgut. Our finding provides new insights into the function of lncRNAs in regulating virus infection in an important insect vector.

长链非编码RNA（long non-coding RNAs，lncRNAs）可通过转录及转录后调控参与多种生物学功能。然而，目前对于其在昆虫介导的病毒传播过程中的功能尚不明晰。本研究借助RNA测序（RNA-Seq）技术，分析了灰飞虱（Laodelphax striatellus，Fallén）中肠响应水稻黑条矮缩病毒（Rice black-streaked dwarf virus，RBSDV）感染时的差异表达lncRNAs。本研究共鉴定得到13927个lncRNAs，其中超过69%的lncRNAs定位于基因间区。在这些lncRNAs中，176个呈现差异表达状态，且预测可靶向调控168个反式调控基因。研究人员选取10个差异表达的lncRNAs，通过实时定量逆转录PCR（RT-qPCR）验证了其表达变化情况。KEGG富集分析结果显示，这些靶基因显著富集于嘌呤代谢、缬氨酸、亮氨酸与异亮氨酸降解以及脂肪酸延伸等关键生物学过程。此外，研究人员还通过RT-qPCR分析了在人乳头瘤病毒感染通路（Human papillomavirus infection pathway）中显著富集的差异表达lncRNAs及其预测靶基因的表达水平。结果表明，部分lncRNAs与其靶基因存在共表达现象。其中，名为MSTRG15394的lncRNA及其编码分泌型蛋白酶抑制剂（secreted protease inhibitor，PI）的靶基因，在RBSDV感染后转录水平显著上调。敲低MSTRG15394的表达可在mRNA水平下调PI的表达量。通过RNA干扰抑制MSTRG15394或PI的表达，均可促进RBSDV在灰飞虱中肠内的积累。本研究为解析长链非编码RNA在重要昆虫媒介调控病毒感染过程中的功能提供了全新的视角。