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WSSV感染对中国明对虾bantam及其候选靶基因的影响
李旭鹏1, 孔杰1,2, 孟宪红1, 栾生1,2, 罗坤1
1.农业农村部海洋渔业可持续发展重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071;2.青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室 青岛 266071
摘要:
Bantam能够调控细胞增殖、细胞凋亡等过程,影响生物的免疫过程。本研究利用实时荧光定量PCR(qRT-PCR)技术对感染WSSV的中国明对虾(Fenneropenaeus chinensis)肝胰腺和鳃组织内bantam表达水平进行检测,发现感染WSSV后6、12、24和48 h,中国明对虾肝胰腺中的bantam表达水平分别是对照组的(0.16±0.03)(P<0.05)、(0.63±0.26)、(0.32±0.06)(P<0.05)和(0.41±0.13)倍;中国明对虾鳃中的bantam表达水平分别是对照组的(0.30±0.17)(P<0.05)、(1.88±0.26)(P<0.01)、(0.84±0.36)和(0.51±0.25)倍。利用miRanda软件进一步对中国明对虾bantam靶基因进行预测分析,评分最高的靶基因是泛素缀合酶E2。中国明对虾泛素缀合酶E2包含UBCc功能域。多序列比对显示,UBCc功能域氨基酸残基序列在不同物种间保守性较高。进化树分析显示,分类学地位相近的物种的泛素缀合酶E2聚为一类。qRT-PCR检测感染WSSV的中国明对虾肝胰腺和鳃中的泛素缀合酶E2表达水平,结果显示,在感染WSSV后6、12、24和48 h,中国明对虾肝胰腺中泛素缀合酶E2的表达水平分别是对照组的(0.54±0.10)、(1.19±0.62)、(3.69±0.51) (P<0.01)和(1.94±0.07)(P<0.05)倍;中国明对虾鳃中泛素缀合酶E2的表达水平分别是对照组的(0.22±0.05)、(1.34±0.38)、(4.29±0.52)(P<0.01)和(1.28±0.79)倍。研究表明,bantam和泛素缀合酶E2的表达都受WSSV侵染的影响,可能与中国明对虾和WSSV之间的互作相关。但bantam和泛素缀合酶E2表达水平的变化是对虾抵抗WSSV侵染过程的免疫反应,还是宿主基因被病毒胁迫后的结果,需要进一步验证。
关键词:  中国明对虾  WSSV  bantam  泛素缀合酶E2  表达
DOI:
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基金项目:
Effect of White Spot Syndrome Virus Infection on bantam and Candidate Target Genes in the Chinese Shrimp Fenneropenaeus chinensis
LI Xupeng1, KONG Jie1,2, MENG Xianhong1, LUAN Sheng1,2, LUO Kun1
1.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071;2.Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266071
Abstract:
Bantam is a type of miRNA involved in cell proliferation and apoptosis, and can affect immune processes in animals. In the present study, bantam expression in the hepatopancreas and gill of WSSV-infected Chinese shrimp Fenneropenaeus chinensis was determined using the qRT-PCR technique. The results showed that bantam expression in the hepatopancreas of Chinese shrimp 6, 12, 24, and 48 h post infection was (0.16±0.03)(P<0.05), (0.63±0.26), (0.32±0.06) (P<0.05), and (0.41±0.13) fold that of the expression in the controls, respectively. Bantam expression in the gill of Chinese shrimp 6, 12, 24, and 48 h post infection was (0.30±0.17) (P<0.05), (1.88±0.26) (P<0.01), (0.84±0.36), and (0.51±0.25) fold that of the expression in the controls, respectively. The target genes of bantam were predicted using the miRanda software. The ubiquitin-conjugating enzyme E2 was the predicted target gene with the highest score. The Chinese shrimp ubiquitin-conjugating enzyme E2 contains the ubiquitin-conjugating enzyme E2, catalytic (UBCc) domain. Multiple alignment results showed that the UBCc domain was relatively conserved among different species. Phylogenetic analysis showed that the ubiquitin-conjugating enzyme E2 in species with closer taxonomic status was clustered in a branch. Pair-wise and multiple sequence alignment revealed that the ubiquitin-conjugating enzyme E2 was relatively well-conserved. Ubiquitin- conjugating enzyme E2 expression in the hepatopancreas and gill of WSSV-infected Chinese shrimp was further determined using qRT-PCR. The results showed that the expression of ubiquitin-conjugating enzyme E2 in the hepatopancreas of Chinese shrimp 6, 12, 24, and 48 h post-WSSV infection was (0.54±0.10), (1.19±0.62), (3.69±0.51)(P<0.01), and (1.94±0.07)(P<0.05) fold that of the expression in the controls, respectively. The expression of ubiquitin-conjugating enzyme E2 in the gill of Chinese shrimp 6, 12, 24 and 48 h post WSSV infection was (0.22±0.05), (1.34±0.38), (4.29±0.52) (P<0.01), and (1.28±0.79) fold that of the expression in the controls. These results suggest that both bantam and the target gene ubiquitin-conjugating enzyme E2 might be involved in the interaction between Chinese shrimp and WSSV. Whether the altered expression of bantam and ubiquitin-conjugating enzyme E2 represents the immune response of shrimps to WSSV infection, or the hijacking of cells by WSSV during the viral infection process, needs further study.
Key words:  Fenneropenaeus chinensis  WSSV  bantam  Ubiquitin-conjugating enzyme E2  Expression