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| 红鳍东方鲀低温胁迫应答主效QTL候选基因的表达特征分析 |
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朱理光,刘志峰,马爱军,王新安,孙志宾,常浩文,刘圣聪,包玉龙,马得友
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1.大连海洋大学水产与生命学院 辽宁 大连 116023;2.中国水产科学研究院黄海水产研究所
山东省海洋渔业生物技术与遗传育种重点实验室 青岛市海水鱼类种子工程与生物技术重点实验室
山东 青岛 266071;3.海洋生物学与生物技术功能实验室 山东 青岛 266071;4.海洋生物学与生物技术功能实验室 山东 青岛 266072;5.海洋生物学与生物技术功能实验室 山东 青岛 266073;6.海洋生物学与生物技术功能实验室 山东 青岛 266074;7.海洋生物学与生物技术功能实验室 山东 青岛 266075;8.唐山牧海水产养殖有限公司 河北 唐山 063200
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| 摘要: |
| 红鳍东方鲀(Takifugu rubripes)为暖温性、广温性鱼类，低温的冬季海水会对其生存造成很大影响，因此，选育具有抗寒性状的品系对其产业发展具有重要意义。本研究在利用QTL (quantitative trait locus)定位分析筛选出6个与红鳍东方鲀耐低温相关的候选基因(tacc2、fsip1、exoc4、arhgap44a、pde10a和unc5b)的基础上，通过Real-time PCR检测这6个基因在低温胁迫下在肝脏、心脏和肾脏中表达量的变化。实验用鱼为课题组建立的同一家系的8月龄鱼，共设置3个温度梯度(8 ℃、13 ℃和18 ℃)，8 ℃和13 ℃为低温组，18 ℃为对照组。结果显示，6个基因在不同温度下的3个组织中均有不同程度的表达。其中，pde10a基因在3个组织中的表达均呈先升高后下降的趋势；tacc2和exoc4基因在8 ℃组肝脏、肾脏以及心脏中的表达分别呈先下降再趋于稳定、先升高再趋于稳定和先上升后下降的趋势；unc5b基因在肝脏和心脏中的表达量较低，在低温组实验前期的肾脏中呈现高表达；arhgap44a基因在肝脏中的表达呈上升趋势，在心脏和肾脏中整体表达无明显变化；fsip1基因在肝脏中的表达呈下降趋势，在心脏和肾脏中的表达呈先上升后下降的趋势。这6个基因在红鳍东方鲀组织中均随着时间和温度变化具有差异性表达，在低温胁迫下呈现积极响应，表明这6个QTL候选基因在红鳍东方鲀低温适应中具有潜在的重要作用。本研究可为红鳍东方鲀耐低温相关信号通路研究以及耐低温品种选育提供理论依据。 |
| 关键词: 红鳍东方鲀 pde10a基因 tacc2基因 unc5b基因 exoc4基因 arhgap44a基因 fsip1基因 低温胁迫 |
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| Evaluating the transcriptional regulation of six major QTL candidate genes during low temperature stress in Takifugu rubripes |
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ZHU Liguang1,2,3,2,4, LIU Zhifeng2,4, MA Aijun5,6, WANG Xin´an7,8, SUN Zhibin9,8, CHANG Haowen10,8, LIU Shengcong11, BAO Yulong12, MA Deyou1
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1.College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China;2.Shandong Key Laboratory of Marine Fisheries Biotechnology and Genetic Breeding, Qingdao Key Laboratory of Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China;3.Laboratory for Marine Biology and Biotechnology, Qingdao 266071, China
College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China;4.Laboratory for Marine Biology and Biotechnology, Qingdao 266071, China;5.Shandong Key Laboratory of Marine Fisheries Biotechnology and Genetic Breeding, Qingdao Key Laboratory of Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266072, China;6.Labora 攈䂞 辠࿅ 攠䂞 辠࿅ 擀䂞 辠࿅ 擠䂞 辠࿅ 摨䂞 辠࿅ 撈䂞 辠࿅ a
;7.Shandong Key Laboratory of Marine Fisheries Biotechnology and Genetic Breeding, Qingdao Key Laboratory of Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266073, China;8.Labora 6 ;9.Shandong Key Laboratory of Marine Fisheries Biotechnology and Genetic Breeding, Qingdao Key Laboratory of Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266074, China;10.Shandong Key Laboratory of Marine Fisheries Biotechnology and Genetic Breeding, Qingdao Key Laboratory of Marine Fish Breeding and Biotechnology, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266075, China;11.Tangshan Muhai Aquaculture Co. Ltd., Tangshan 063200, China;12.Tangshan Muhai Aquaculture Co. Ltd., Tangshan 063201, China
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| Abstract: |
| Takifugu rubripes are warm temperate fish, suggesting that the reduced seawater temperatures in winter are likely to have a substantial impact on their survival. Considering this, there is likely to be some industrial value in breeding extremely low-temperature tolerant varieties of this fish. Here, we evaluate the expression changes in six major QTL candidate genes (tacc2, fsip1, exoc4, arhgap44a, pde10a, and unc5b) in response to reduced temperature in an effort to understand cold tolerance in T. rubripes. The expression changes of these six genes in the liver, heart, and kidney were detected using real-time quantitative PCR. This study used three groups of 8-month-old fish, all from the same family established by our research group, exposed to three different temperature gradients, where 8 ℃ and 13 ℃ acted as the minimum in the low temperature groups and 18 ℃ acted as the minimum in the control group. Our results showed that all six genes were expressed at different levels across each of these three tissues at different temperatures. The relative expression of pde10a first increased and then decreased in all three tissues, whereas the relative expression of tacc2 and exoc4 were distinctly different in the liver, kidney, and heart at 8 ℃. In this case, these transcripts first decreased and then stabilized in the liver, increased and then stabilized in the kidneys, and increased and then decreased in the heart. The relative expression of unc5b was low in the liver and heart, but high in the kidney following a second week of low-temperature growth, whereas arhgap44a expression was slightly upregulated in the liver and stable in the kidney and heart. fsip1 expression demonstrated a downward trend in the liver but seemed to first increase and then decrease in the heart and kidney. Taken together, these results demonstrate that all six of these genes are differentially expressed in different tissues of T. rubripes, with these differences exhibiting dynamic changes with respect to tissue origin and temperature. In addition, this data clearly revealed a positive correlation between cold stress and the expression of these QTL candidate genes. Thus, we can conclude that these six QTL candidate genes may play a substantial role in the low temperature adaptation of T. rubripes. This is significant because although low temperature is known to be an important factor limiting the development of the industrial utility of T. rubripes, there are still relatively few reports describing their cold stress response. This study provides a theoretical basis for the study of signaling pathways related to the low temperature tolerance response of T. rubripes and the development of low temperature tolerant varieties. |
| Key words: Takifugu rubripes pde10a tacc2 unc5b exoc4 arhgap44a fsip1 Low temperature stress |
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