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脊尾白虾对低氧响应的转录组学分析
曹梅,王兴强,秦传新,沈晔,张子杨,钱诗悦
1.江苏海洋大学海洋科学与水产学院 连云港 222005;2.中国水产科学研究院南海水产研究所 广东省渔业生态环境重点实验室 广州 510300
摘要:
本研究通过高通量测序,分析低氧胁迫下脊尾白虾(Palaemon carincauda)某些基因的差异表达,获得10.62 Gb高质量测序数据,组装得到155113条转录本和118953条Unigene。注释Unigene 37580条。其中,33659条Unigene与Nr蛋白数据库基因同源;11275条Unigene注释到KEGG数据库,归类到223个代谢通路。低氧胁迫产生1392条差异表达基因,包括311条上调基因和1081条下调基因,784条差异基因得到注释,并富集到抗氧化活性、细胞连接、蛋白结合转录因子活性、多细胞生物过程、复制和生殖等过程,表明低氧胁迫激活了虾体适应缺氧的一系列生理活动。其中,低氧胁迫下,低氧诱导因子1(HIF1) 2个亚基HIF1α和HIF1β表达量上调;实时定量测定证实,在胁迫的后期,脊尾白虾肝胰脏和鳃HIF1α和HIF1β明显上调,推测脊尾白虾细胞在低溶氧环境下诱导HIF产生,刺激机体增加血液氧的供应能力。同时,低氧胁迫下,脊尾白虾差异基因富集到糖酵解/葡萄糖生成、精氨酸和脯氨酸代谢和丙酮酸代谢等通路,表明虾体缺氧使糖酵解等无氧代谢途径增强,同时促进了部分糖类和氨基酸的代谢。另外,低氧胁迫下,脊尾白虾溶酶体通路、吞噬通路、过氧化物酶体通路和内吞作用通路的差异基因较多,推测低氧诱导因子可能通过抑制线粒体生物合成和活化线粒体自噬来降低线粒体氧耗。
关键词:  脊尾白虾  低氧胁迫  转录组分析  差异基因表达
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Transcriptome analysis of Palaemon carincauda subject to hypoxic stress
CAO Mei1, WANG Xingqiang1, QIN Chuanxin2,3, SHEN Ye1, ZHANG Ziyang1, QIAN Shiyue1
1.College of Marine Science and Fisheries, Jiangsu Ocean University, Lianyungang 222005;2.Guangdong Provincial Key Laboratory of Fishery Ecology and Environment;3.South China Sea Fisheries Research Institute, Chinese Academy of Fisheries Sciences, Guangzhou 510300
Abstract:
A rapid reduction in the dissolved oxygen concentration of water is an important factor contributing to disease in shrimps. In this study, we examined differential gene expression in the shrimp Palaemon carincauda under conditions of hypoxic stress. We obtained 10.62 Gb of high-quality sequencing data, from which 155113 transcripts and 118953 unigenes were assembled. Among the unigenes, 37580 were annotated and 33659 were found to be homologous to genes in the Nr protein database. We also annotated 11275 unigenes using the KEGG database, which were further classified into 223 metabolic pathways. We detected 1392 genes that were differentially expressed in shrimps exposed to hypoxic stress, among which 311 and 1081 were up- and down-regulated, respectively, and 784 were annotated. The enrichment of differentially expressed genes in antioxidant activity, cell connection, protein-binding transcription factor activity, multicellular biological processes, replication, and reproductive processes indicated that exposure to hypoxia activated a series of physiological responses in shrimps associated with adaptation to low levels of dissolved oxygen. Among the genes up-regulated under hypoxic stress was hypoxic induction factor 1 (HIF1), which is comprised the two subunits HIF1α and HIF1β. RT-PCR analysis revealed that during the latter stages of hypoxic stress, there was a notable up-regulated expression of HIF1α and HIF1β in the hepatopancreas and gills of P. carincauda. These observations indicated that P. carincauda cells induced hypoxic induction factor production in a hypoxic environment, thereby inducing an increase in blood oxygen supply. We also detected an enrichment of differentially expressed genes in the glycolysis/glucose generation pathway, arginine and proline metabolism, and pyruvate metabolism in response to hypoxic stress, which indicated an upregulation of anaerobic metabolic processes such as glycolysis, and increased metabolism of certain carbohydrates and amino acids. In addition, we detected numerous differentially expressed genes associated with the pathways involving lysozymes, phagocytosis, peroxisomes, and endocytosis in P. carincauda exposed to hypoxia, thereby indicating that HIFs might reduce mitochondrial oxygen consumption by inhibiting mitochondrial biosynthesis and activating mitochondrial autophagy.
Key words:  Palaemon carincauda  Stress hypoxic  Transcriptome analysis  Differential gene expression