侵染意大利蜜蜂工蜂的东方蜜蜂微孢子虫的长链非编码RNA差异表达谱及其调控作用
首发时间:2020-04-23
摘要:本研究基于前期获得的东方蜜蜂微孢子虫(Nosema ceranae)感染7 d和10 d的意大利蜜蜂(Apis mellifera ligustica,简称意蜂)工蜂中肠转录组数据筛滤得到病原(NcT1L和NcT2L)的lncRNA组学数据,并结合东方蜜蜂微孢子虫的纯净孢子(NcCKL)的lncRNA组学数据进行比较分析,在NcCKL、NcT1L和NcT2L组分别鉴定出26、23和22条lncRNA,共有lncRNA为21条。通过RT-PCR验证了6条lncRNA的真实表达。共有5条lncRNA可作为反义lncRNA结合相应的mRNA。从NcCKL vs NcT1L、NcCKL vs NcT2L和NcT1L vs NcT2L比较组分别筛选出19、24和4个差异表达lncRNA(DElncRNA)。上述DElncRNA分别调控26、27和2个上下游基因,分别涉及12、6和0个功能条目,以及17、18和0条通路。上述DElncRNA可通过反式作用分别调控30、39和8个mRNA,多数可同时调控1个正相关的mRNA和1个负相关的mRNA。NcCKL vs NcTL1中的15个DElncRNA可靶向195个miRNA及204个mRNA ,NcCKL vs NcT2L中的23个DElncRNA可靶向211个miRNA及216个mRNA,NcT1L vs NcT2L中的4个DElncRNA可靶向94个miRNA及73个mRNA。通过RT-qPCR验证了6条DElncRNA的差异表达和测序数据的可靠性。研究结果提供了东方蜜蜂微孢子虫侵染意蜂工蜂过程的lncRNA差异表达谱,揭示了DElncRNA在病原侵染过程的潜在功能,为侵染机制的阐明提供了基础。
关键词: 东方蜜蜂微孢子虫 意大利蜜蜂 长链非编码RNA 侵染机制 调控网络
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Dfferential expression pattern and regulatory function of long non-coding RNAs in Nosema ceranae infecting Apis mellifera ligustica worker
Abstract:In this study, long non-coding RNA (lncRNA) omics dataset of Nosema ceranae within infection process was filtered out based on our previously obtained transcriptime dataset from Apis mellifera ligustica workers\' midguts at 7 d and 10 d post N. ceranae infection, and comparative analysis was performed combined with lncRNA omics dataset of clean spores of N. ceranae. In NcCKL, NcT1L and NcT2L groups, 26, 23 and 22 lncRNAs were respectively identified, and the number of shared lncRNAs was 21. The true expression of six lncRNAs was validated using RT-PCR. Additionally, five lncRNAs as antisense lncRNAs can link to corresponding mRNAs. Further, 19, 24 and four differentially expressed lncRNAs (DElncRNAs) were identified in NcCKL vs NcT1L, NcCKL vs NcT2L and NcT1L vs NcT2L comparison groups, respectively, and these DElncRNAs can respectively regulate 17, 18 and zero pathways. In addition, the aformentioned DElncRNAs could regulate 30, 39 and 8 mRNAs via trans-acting manner, and majority of DElncRNAs can simutaneously regulated one positively-related and one negatively-related mRNAs. Moreover, 15 DElncRNAs in NcCKL vs NcT1L can target 195 miRNAs and 204 mRNAs; 23 DElncRNAs in NcCKL vs NcT2L can target 211 miRNAs and 216 mRNAs; four DElncRNAs in NcT1L vs NcT2L can target 94 miRNAs and 73 mRNAs. Finally, the differential expression of six DElncRNAs and reliability of sequencing data was verified using RT-qPCR. The results provide the differential expression pattern of lncRNAs in N. ceranae during the infection process, reveal the potential function of DElncRNAs in N. ceranae infecting A. m. ligustica worker, and lay a foundation for clarification of the infection mechanism.
Keywords: Nosema ceranae Apis mellifera ligustica long non-coding RNA infection mechanism regulatory network
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侵染意大利蜜蜂工蜂的东方蜜蜂微孢子虫的长链非编码RNA差异表达谱及其调控作用
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