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罗氏沼虾(giant fresh water prawn)幼体转录组分析对罗氏沼虾诺达病毒(MrNV)感染的反应:从头组装和功能注释。

Transcriptomic analysis of Macrobrachium rosenbergii (giant fresh water prawn) post-larvae in response to M. rosenbergii nodavirus (MrNV) infection: de novo assembly and functional annotation.

机构信息

Department of Biology, Srinakharinwirot University, Sukhumvit 23, Bangkok, 10110, Thailand.

QCPU, Queen's Cardiopulmonary Unit, Translational Institute of Medicine (TIME), Department of Medicine, Queen's University, Kingston, ON, K7L 3N6, Canada.

出版信息

BMC Genomics. 2019 Oct 22;20(1):762. doi: 10.1186/s12864-019-6102-6.

Abstract

BACKGROUND

Macrobrachium rosenbergii, is one of a major freshwater prawn species cultured in Southeast Asia. White tail disease (WTD), caused by Macrobrachium rosenbergii nodavirus (MrNV), is a serious problem in farm cultivation and is responsible for up to 100% mortality in the post larvae stage. Molecular data on how M. rosenbergii post-larvae launches an immune response to an infection with MrNV is not currently available. We therefore compared the whole transcriptomic sequence of M. rosenbergii post-larvae before and after MrNV infection.

RESULTS

Transcriptome for M. rosenbergii post-larvae demonstrated high completeness (BUSCO Complete: 83.4%, fragmentation: 13%, missing:3.3%, duplication:16.2%; highest ExN50 value: 94%). The assembled transcriptome consists of 96,362 unigenes with N of 1308 bp. The assembled transcriptome was successfully annotated against the NCBI non-redundant arthropod database (33.75%), UniProt database (26.73%), Gene Ontology (GO) (18.98%), Evolutionary Genealogy of Genes: Non-supervised Orthologous Groups (EggNOG) (20.88%), and Kyoto Encyclopedia of Genes and Genome pathway (KEGG) (20.46%). GO annotations included immune system process, signaling, response to stimulus, and antioxidant activity. Differential abundance analysis using EdgeR showed 2413 significantly up-regulated genes and 3125 significantly down-regulated genes during the infection of MrNV.

CONCLUSIONS

This study reported a highly complete transcriptome from the post-larvae stage of giant river prawn, M. rosenbergii. Differential abundant transcripts during MrNV infection were identified and validated by qPCR, many of these differentially abundant transcripts as key players in antiviral immunity. These include known members of the innate immune response with the largest expression change occurring in the M. rosenbergii post-larvae after MrNV infection such as antiviral protein, C-type lectin, prophenol oxidase, caspase, ADP ribosylation factors, and dicer.

摘要

背景

罗氏沼虾是东南亚主要养殖的淡水虾品种之一。白尾病(WTD)由罗氏沼虾诺达病毒(MrNV)引起,是养殖中的严重问题,在幼虾后期可导致高达 100%的死亡率。目前尚无罗氏沼虾幼虾感染 MrNV 后如何启动免疫反应的分子数据。因此,我们比较了 MrNV 感染前后罗氏沼虾幼虾的全转录组序列。

结果

罗氏沼虾幼虾的转录组具有很高的完整性(BUSCO 完整度:83.4%,片段化:13%,缺失:3.3%,重复:16.2%;最高 ExN50 值:94%)。组装的转录组由 96362 个 unigene 组成,N 为 1308bp。组装的转录组成功地注释了 NCBI 非冗余节肢动物数据库(33.75%)、UniProt 数据库(26.73%)、基因本体论(GO)(18.98%)、非监督同源群基因进化(EggNOG)(20.88%)和京都基因与基因组百科全书途径(KEGG)(20.46%)。GO 注释包括免疫系统过程、信号转导、对刺激的反应和抗氧化活性。使用 EdgeR 进行差异丰度分析显示,MrNV 感染期间有 2413 个显著上调基因和 3125 个显著下调基因。

结论

本研究报道了罗氏沼虾幼虾阶段高度完整的转录组。通过 qPCR 验证了 MrNV 感染过程中差异丰度的转录本,其中许多差异丰度的转录本是抗病毒免疫的关键因子。这些包括先天免疫反应的已知成员,其中 MrNV 感染后罗氏沼虾幼虾中表达变化最大的是抗病毒蛋白、C 型凝集素、原酚氧化酶、半胱天冬酶、ADP 核糖基化因子和 dicer。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/912d/6805343/34cf51351dd9/12864_2019_6102_Fig1_HTML.jpg

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