Integrated analysis of mRNA-seq in the haemocytes of Eriocheir sinensis in response to Spiroplasma eriocheiris infection
Integrated analysis of mRNA-seq in the haemocytes of Eriocheir sinensis in response to Spiroplasma eriocheiris infection
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中华绒螯蟹血细胞响应绒毛虫感染的 mRNA-seq 整合分析
DOI：
10.1016/j.fsi.2017.07.036
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发表时间：
2017
影响因子：
4.7
通讯作者：
Meng Qingguo
中科院分区：
文献类型：
--
作者：
Wang Yinghui;Xiu Yunji;Bi Keran;Ou Jiangtao;Gu Wei;Wang Wen;Meng Qingguo
中华绒螯蟹（Eriocheir sinensis）是一种重要的经济甲壳动物，但其病害严重。从患震颤病的中华绒螯蟹（E. sinensis）中分离到的螺旋体（Spiroplasmaeriocheiris）是首次发现的淡水甲壳动物致死性病原体。为了了解中华绒螯蟹志贺菌的发病机制，使用Illumina HiSeq 2500获得了注射后1 d和7 d实验组和对照组血细胞的转录组学谱。结果表明，DZ 1（对照组）、DZ 7（对照组）、SY 1（实验组）和SY 7（实验组）的cDNA文库分别获得40，358，724、44，462，112、45，516，576和37，713，728个双端clean reads。总共组装了106，641个独特的转录片段（unigenes），平均长度为710 bp。在刺激的第一天，与对照组相比，实验组中有33，084个上调转录本和19，208个下调转录本。在刺激第7天，实验组与对照组相比，共检测到40,198个上调转录本和12,032个下调转录本。通过KEGG途径分析鉴定了一些典型的免疫相关途径，包括补体和凝血级联、VEGF信号传导途径、Wnt信号传导途径、自然杀伤细胞介导的细胞毒性、MAPK信号传导途径、神经活性配体-受体相互作用和溶酶体途径。我们在KEGG通路中发现了重要的免疫相关基因（GNPTAB、MASP 2、F7、F5、NFATC、TRAF 6、MAP 3 K5和TRa），并通过qRT-PCR分析证实了这些基因。此外，显着丰富的神经活性配体-受体相互作用途径与强烈的阵发性震颤的感染螃蟹。本研究结果为进一步分析中华绒螯蟹防御绒螯蟹入侵的机制提供了有价值的信息。
The Chinese mitten crabEriocheir sinensisis an important economic crustacean that has been exposed to various diseases. Spiroplasmaeriocheiris, isolated from tremor-diseasedE. sinensis, was first identified as a lethal pathogen of freshwater crustaceans. To understand the pathogenesis ofS. eriocheiristoE. sinensis, the transcriptomic profiles of haemocytes in the experimental and control groups at 1 d and 7 d post-injection were obtained using Illumina HiSeq 2500. These results showed that 40,358,724, 44,462,112, 45,516,576 and 37,713,728 paired-end clean reads were obtained from the cDNA libraries of DZ1 (the control group at 1 d), DZ7 (the control group at 7 d), SY1 (the experimental group at 1 d) and SY7 (the experimental group at 7 d), respectively. In total, 106,641 unique transcript fragments (unigenes) were assembled, with an average length of 710 bp. On the first day of stimulation, 33,084 up-regulated transcripts and 19,208 down-regulated transcripts were found in the experimental group compared with those in the control group. On the seventh day of stimulation, 40,198 up-regulated transcripts and 12,032 down-regulated transcripts were found in the experimental group compared with those in the control group. Some canonical immune-related pathways were identified via KEGG pathway analysis, including complement and coagulation cascades, the VEGF signalling pathway, the Wnt signalling pathway, natural killer cell-mediated cytotoxicity, the MAPK signalling pathway, neuroactive ligand-receptor interactions, and the Lysosome pathway. We found important immune-related genes (GNPTAB, MASP2, F7, F5, NFATC, TRAF6, MAP3K5, and TRa) in the KEGG pathway, and those genes were confirmed by qRT-PCR analysis. In addition, the significantly enriched neuroactive ligand-receptor interaction pathway was associated with intense paroxysmal tremors of infected crabs. Our results provide valuable information for the further analysis of the mechanisms ofE. sinensisdefence againstS. eriocheirisinvasion.