Jiangsu Key Laboratory of Biochemistry and Biotechnology of Marine Wetland, School of Marine and Biological Engineering, Yancheng Institute of Technology, Yancheng, 224051, Province Jiangsu, China.
Jiangsu Key Laboratory of Biochemistry and Biotechnology of Marine Wetland, School of Marine and Biological Engineering, Yancheng Institute of Technology, Yancheng, 224051, Province Jiangsu, China.
Fish Shellfish Immunol. 2021 Dec;119:651-669. doi: 10.1016/j.fsi.2021.11.002. Epub 2021 Nov 4.
Macrobrachium rosenbergii (M. rosenbergii), is a major aquaculture species in China and Southeast Asia. However, infection with Spiroplasma eriocheiris (S. eriocheiris) has caused huge economic losses to the cultivation of M. rosenbergii. Currently, there are few reports on the immune response mechanism of M. rosenbergii that are infected with S. eriocheiris. To clarify the immune response mechanism of M. rosenbergii infected with S. eriocheiris, the key immune genes which respond to the infection with the pathogen and the regulation of related microRNAs (miRNAs) on them were identified. In this study, the mRNA and miRNA transcriptome of hepatopancreas of M. rosenbergii at different infection stages were analyzed using high-throughput sequencing and qRT-PCR. In the mRNA transcriptome, 27,703 and 33,402 genes were expressed in healthy and susceptible M. rosenbergii, respectively. By digital gene-expression profiling analysis, 23,929 and 24,325 genes were expressed, and 223 and 373 genes were significantly up-regulated and down-regulated, respectively. A total of 145 key genes related to Toll, IMD, JAK/STAT and MAPK were excavated from the transcriptome. In the miRNA transcriptome, 549 miRNAs (Conserved: 41, PN-type: 83, PC-type: 425) were sequenced, of which 87 were significantly up-regulated and 23 were significantly down-regulated. Among the related immune pathways, there are 259 miRNAs involved in the regulation of target genes in the Toll and IMD pathways, 231 JAK/STAT pathways and 122 MAPK pathways. qRT-PCR differential detection of immune-related miRNAs and mRNAs showed that 22 miRNAs with significant differences (P < 0.05) such as mro-miR-100, PC-mro-3p-27 and PN-mro-miR-316 had corresponding regulatory relationships with 22 important immune genes such as TLR2, TLR3, TLR4, TLR5, MyD88, Pelle and Relish in different stages after infection. In this study, the immune genes and related regulatory miRNAs of M. rosenbergii in response to S. eriocheiris infection were obtained. The results can provide basic data to further reveal the immune defense mechanism of M. rosenbergii against S. eriocheiris infection.
罗氏沼虾（Macrobrachium rosenbergii）是中国和东南亚地区主要的水产养殖品种。然而，感染沼虾螺旋体（Spiroplasma eriocheiris）会给罗氏沼虾的养殖带来巨大的经济损失。目前，关于感染沼虾螺旋体的罗氏沼虾的免疫反应机制的报道较少。为了阐明罗氏沼虾感染沼虾螺旋体的免疫反应机制，本研究鉴定了对病原体感染有反应的关键免疫基因及其相关微小 RNA（miRNA）的调控。本研究采用高通量测序和 qRT-PCR 分析了罗氏沼虾在不同感染阶段的肝胰腺的 mRNA 和 miRNA 转录组。在 mRNA 转录组中，健康罗氏沼虾和易感罗氏沼虾分别表达了 27703 个和 33402 个基因。通过数字基因表达谱分析，分别有 23929 个和 24325 个基因表达，分别有 223 个和 373 个基因显著上调和下调。从转录组中挖掘出与 Toll、IMD、JAK/STAT 和 MAPK 相关的 145 个关键基因。在 miRNA 转录组中，共测序到 549 个 miRNA（保守型：41 个，PN 型：83 个，PC 型：425 个），其中 87 个 miRNA 显著上调，23 个 miRNA 显著下调。在相关免疫途径中，有 259 个 miRNA 参与 Toll 和 IMD 途径、231 个 JAK/STAT 途径和 122 个 MAPK 途径中靶基因的调控。免疫相关 miRNA 和 mRNA 的 qRT-PCR 差异检测显示，在感染后不同阶段，22 个差异显著（P<0.05）的 miRNA，如 mro-miR-100、PC-mro-3p-27 和 PN-mro-miR-316，与 22 个重要免疫基因，如 TLR2、TLR3、TLR4、TLR5、MyD88、Pelle 和 Relish 之间存在相应的调控关系。本研究获得了罗氏沼虾对沼虾螺旋体感染的免疫基因和相关调控 miRNA，可为进一步揭示罗氏沼虾对沼虾螺旋体感染的免疫防御机制提供基础数据。
