留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码
高阳, 王超宇, 刘鑫, 梁夏颖, 赵哲, 史燕. 暗纹东方鲀耐寒相关基因CIRBPHMGB1AFP-的分子特征和对低温胁迫的响应[J]. 水生生物学报, 2023, 47(5): 756-766. DOI: 10.7541/2023.2022.0239
引用本文: 高阳, 王超宇, 刘鑫, 梁夏颖, 赵哲, 史燕. 暗纹东方鲀耐寒相关基因CIRBPHMGB1AFP-的分子特征和对低温胁迫的响应[J]. 水生生物学报, 2023, 47(5): 756-766. DOI: 10.7541/2023.2022.0239
GAO Yang, WANG Chao-Yu, LIU Xin, LIANG Xia-Ying, ZHAO Zhe, SHI Yan. MOLECULAR CHARACTERIZATION OF COLD-TOLERANCE RELATED GENES CIRBP, HMGB1, AFP- FROM TAKIFUGU OBSCURUS AND THEIR RESPONSE TO LOW TEMPERATURE STRESS[J]. ACTA HYDROBIOLOGICA SINICA, 2023, 47(5): 756-766. DOI: 10.7541/2023.2022.0239
Citation: GAO Yang, WANG Chao-Yu, LIU Xin, LIANG Xia-Ying, ZHAO Zhe, SHI Yan. MOLECULAR CHARACTERIZATION OF COLD-TOLERANCE RELATED GENES CIRBP, HMGB1, AFP- FROM TAKIFUGU OBSCURUS AND THEIR RESPONSE TO LOW TEMPERATURE STRESS[J]. ACTA HYDROBIOLOGICA SINICA, 2023, 47(5): 756-766. DOI: 10.7541/2023.2022.0239

暗纹东方鲀耐寒相关基因CIRBPHMGB1AFP-的分子特征和对低温胁迫的响应

MOLECULAR CHARACTERIZATION OF COLD-TOLERANCE RELATED GENES CIRBP, HMGB1, AFP- FROM TAKIFUGU OBSCURUS AND THEIR RESPONSE TO LOW TEMPERATURE STRESS

  • 摘要: 为了探索暗纹东方鲀(Takifugu obscurus)对低温环境的响应机制, 克隆了暗纹东方鲀耐寒相关基因CIRBPHMGB1AFP-Ⅳ的cDNA序列, 并进行了基因的分子特征和功能分析。组织分布检测显示CIRBPHMGB1在下丘脑、肝脏和肌肉中具有高表达, 而AFP-Ⅳ则主要在肝脏中表达。在受到低温胁迫后, 3种基因在肝脏和下丘脑中的表达呈现不同的变化趋势, 其中CIRBP基因在肝脏中于48h表达量显著增加, 在下丘脑中于12h和48h有上调表达; HMGB1基因在肝脏中呈现逐渐上升的趋势, 于48h达到最大值, 而在下丘脑中呈现先上升后下降的趋势, 处理后2h达到最大, 2—8h下降, 于8h下降至最低, 随后恢复至初始水平; 肝脏中的AFP-Ⅳ在0—24h无显著变化, 在48h上升至最大值。进一步通过大肠杆菌原核表达系统研究了AFP-Ⅳ的抗冻功能, 发现AFP-Ⅳ融合蛋白在–80℃下具有抗冻活性, 并且抗冻活性随着浓度的增加而提高。研究结果表明3种基因都参与了暗纹东方鲀对低温胁迫的应答过程, 为深入探索暗纹东方鲀的耐低温机制奠定了基础。

     

    Abstract: Obscure puffer (Takifugu obscurus) is a euryhaline fugu species and belongs to the Family Tetraodontidae of teleost fish. In China, the Takifugu species is very popular because of its delicious taste and high nutrition, and it is widely farmed on a commercial scale. In recent years, the aquaculture area for obscure puffer is increasing year by year, however, the industry is confronted with many challenges, such as outbreak of diseases and overwintering culture. When winter comes, the growth and breeding of obscure puffer are affected by the environmental temperature. Furthermore, the dramatic decrease of water temperature in winter will cause mass mortality of obscure puffer, and then huge economic losses. Therefore, it is an urgent need for the further development of the industry to obtain low-temperature tolerance fish species through artificial breeding methods, and the premise of this work is to analyze the low-temperature tolerance mechanism of obscure puffer. To explore the response mechanism of obscure puffer to low temperature environment, we cloned the cDNA sequences of tolerance related genes CIRBP, HMGB1 and AFP-Ⅳ and then characterized their molecular features and potential functions. Real-time PCR analysis showed that CIRBP and HMGB1 were highly expressed in the hypothalamus, liver and muscle, while AFP-Ⅳ was mainly expressed in the liver. The obscure puffer was subjected to acute cold stress, then tissue samples were taken at different time periods. Real-time PCR analysis showed that the expression patterns of the above three genes were different in liver and hypothalamus. For example, the expression of CIRBP increased significantly at 48h in liver, while only slightly increased at 12h and 48h in hypothalamus. The expression of HMGB1 showed a gradual increase and reached the highest level at 48h in liver, while it first increased and then decreased in hypothalamus tissue. It reached the maximum at 2h after treatment, decreased at 2—8h, and reached the lowest at 8h, then recovered to the initial level. The expression of AFP-Ⅳ in liver showed no significant change from 0 to 24h, but increased to the maximum at 48h. The antifreeze role of AFP-Ⅳ was further investigated by heterologous expression in Escherichia coli. It was found that the expression of AFP-Ⅳ in E. coli exerted antifreeze effect at –80℃, and the antifreeze activity became apparent with the increase of the concentration of AFP-Ⅳ. Collectively, the results showed that the three genes were involved in the response to low temperature stress, which laid a foundation for further exploring the mechanism of low temperature tolerance in obscure puffer.

     

/

返回文章
返回