患铁虾综合征罗氏沼虾眼柄转录组研究

Study on the Eyestalk Transcriptome of Macrobrachium rosenbergii Suffering from Iron Prawn Syndrome

中文关键词: 罗氏沼虾  铁虾综合征  眼柄  转录组

英文关键词: Macrobrachium rosenbergii  Iron prawn syndrome  Eyestalk  Transcriptome

中文摘要:

      为探索罗氏沼虾(Macrobrachium rosenbergii)铁虾综合征(IPS)的分子机制，采用高通量测序平台(Illumina Hiseq-2500)分别对患IPS罗氏沼虾(IPS虾)和正常罗氏沼虾开展转录组测序，进行生物信息学分析。结果显示，高通量测序共获得56.42 G高质量数据，拼接后得到221 901条单基因序列(unigene)，长度范围为201~30 985 bp，平均长度为1572 bp，N50长度为2867 bp，N90长度为646 bp。将单基因序列分别在Nr、Nt、Swissprot、KEGG、KOG、GO、PFAM数据库进行序列比对及功能注释，103 570条得到注释，其中，GO数据库注释到的单基因序列最多。差异表达分析显示，2003个基因在IPS虾眼柄中差异表达，包括1209个上调基因和794个下调基因，516个基因被注释到242条KEGG通路中，翻译、信号转导和免疫系统富集的差异基因数目最多。催乳素、雌激素、胰岛素、促性腺激素释放激素、胰高血糖素、催产素、谷氨酸能突触、血清素能突触等与生殖调控相关激素的代谢过程在IPS虾与正常虾眼柄之间存在差异。此外，一些已被证明在免疫反应中起重要作用的基因在IPS虾眼柄中显著上调，如血管内皮生长因子受体1、丝氨酸蛋白酶抑制剂6、C型凝集素、芳基硫酸酯酶B、酚氧化酶原激活酶2a、组织蛋白酶B、组织蛋白酶L、甲壳类抗菌肽4等。同时，注释到溶酶体、吞噬体、抗原处理与呈递、细胞凋亡、内吞作用等多条与免疫相关的途径，支持近期研究得出的罗氏沼虾IPS与病原感染相关的结论。本研究为解析罗氏沼虾IPS的成因和分子机制提供了数据支撑。

英文摘要:

      Since 2010, iron prawn syndrome (IPS), characterized by sexual precocity and slow growth, has seriously affected the development of Macrobrachium rosenbergii aquaculture. To investigate the molecular mechanism of IPS, we performed transcriptome sequencing of M. rosenbergii with and without IPS using next-generation sequencing technology. This resulted in a total of 56.42 G of high-quality data being obtained. A total of 221 901 unigenes were obtained, and the length range was 201~30 985 bp. The average length was 1572 bp. The length of N50 was 2867 bp, and the length of N90 was 646 bp. Sequence alignment and functional annotation of the unigenes were conducted in the Nr, Nt, Swissprot, KEGG, KOG, GO, and PFAM databases. A total of 103 570 unigenes were annotated, and the GO database had the most annotated unigenes. The differential expression analysis revealed that 2003 genes, including 1209 upregulated genes and 794 downregulated genes, were differentially expressed in the eyestalks of M. rosenbergii with IPS. Consequently, 516 genes were mapped into 242 KEGG pathways, and the number of differentially expressed genes in translation, signal transduction, and the immune system was the largest. Many reproduction-related pathways were identified, including prolactin, estrogen, insulin, gonadotropin-releasing hormone (GnRH), glucagon, oxytocin, glutamatergic synapses, serotonergic synapses, and other metabolic processes related to reproductive regulation. In addition, some differentially expressed genes that have been proven to play an essential role in the immune response were upregulated in the eyestalks of M. rosenbergii with IPS, such as vascular endothelial growth factor receptor 1, serine proteinase inhibitor 6, C-type lectin, arylsulfatase B, prophenoloxidase-activating enzyme 2a, cathepsin B, cathepsin L, and crustin 4. At the same time, several immune-related pathways, such as lysosome, phagosome, antigen processing and presentation, apoptosis, endocytosis, and many others, were annotated. These results support the conclusion from the recent study that a pathogen is associated with IPS in M. rosenbergii. This study provides a foundation for understanding the causes and molecular mechanisms of IPS in M. rosenbergii.