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徐娴,何琳,林志华,陈铭.2020.盐度胁迫下缢蛏渗透压变化及 V-ATPase H基因的表达分析.动物学杂志,55(5):606-613.
盐度胁迫下缢蛏渗透压变化及 V-ATPase H基因的表达分析
Effects of Salinity Stress on V-ATPase H Expression, Enzyme Activity and Osmotic Pressure in Sinonovacula constricta
投稿时间:2019-10-28  修订日期:2020-08-28
DOI:10.13859/j.cjz.202005009
中文关键词:  缢蛏  V-ATPase H  盐度胁迫  渗透压
英文关键词:Sinonovacula constricta  V-ATPase H  Salt stress  Osmotic pressure
基金项目:国家自然科学基金项目(No. 31802322),宁波市2015年度科技富民项目(No. 2015C10008),浙江省农业新品种选育重大科技专项(No. 2016C02055-9),宁波市一流学科“环境科学与工程”项目
作者单位E-mail
徐娴 浙江万里学院生物与环境学院浙江省水产种质资源高效利用技术研究重点实验室 宁波 315100 915350526@qq.com 
何琳 浙江万里学院生物与环境学院浙江省水产种质资源高效利用技术研究重点实验室 宁波 315100 hlwithyou@qq.com 
林志华 浙江万里学院生物与环境学院浙江省水产种质资源高效利用技术研究重点实验室 宁波 315100 浙江万里学院宁海海洋生物种业研究院 宁波 315100 zhihua9988@126.com 
陈铭 浙江万里学院生物与环境学院浙江省水产种质资源高效利用技术研究重点实验室 宁波 315100 宁波大学海洋学院 宁波 315100 1257620359@qq.com 
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中文摘要:
      为研究V-ATPase H基因在缢蛏(Sinonovacula constricta)盐度胁迫中的功能,以缢蛏成体为实验材料,将缢蛏置于5、15、20、25、35盐度水体中进行胁迫实验,测定了不同胁迫时间缢蛏的血清渗透压、V-ATPase活性变化,克隆了V-ATPase H基因的开放阅读框(ORF)全长序列,并分析其mRNA表达特征。结果显示,低盐组(盐度5和盐度15)和高盐组(盐度25和盐度35)缢蛏血清渗透压变化明显,与对照组(盐度20)有极显著差异(P < 0.01)。随着时间的推移,实验组V-ATPase活力整体呈现先下降后上升的趋势,对照组(盐度20)无明显变化。V-ATPase H基因开放阅读框长度1 440 bp,编码479个氨基酸。qPCR结果显示,V-ATPase H基因在缢蛏鳃中的表达量极显著高于水管、外套膜、肾、肝胰腺、唇瓣、足6个组织(P < 0.01);盐度胁迫下各个实验组V-ATPase H基因在鳃中的表达量持续上升,在24 h达到峰值,显著高于对照组(P < 0.05)。实验结果表明,缢蛏V-ATPase H基因在盐度适应过程中主要在低盐和高盐环境下起到维持自身血清渗透压与外界渗透压平衡的作用。
英文摘要:
      In order to study the function V-ATPase H gene expression on salinity stress in Sinonovacula constricta, the adults were used as experimental materials, and the stress was tested in different salinities (5, 15, 20, 25, 35). The freezing point osmometer was used to determine the difference in serum osmotic pressure, and V-ATPase ELISA detection kit was used to determine the activity of V-ATPase. The full-length ORF sequence of V-ATPase H gene was closed and its mRNA expression was analyzed. The results showed that the serum osmolarity of the low-salt group (salinity 5, salinity 15) and that of high-salt group (salinity 25, salinity 35) changed significantly, both were very significantly different from the control group (salinity 20) (P < 0.01). With the treatment time, the V-ATPase activity of the experimental group showed a decreasing first and then increasing trend, while the control group showed no significant change. The open reading frame (ORF) of the V-ATPase H gene was 1 440 bp in length, encoding 479 amino acids. The qPCR results showed that the expression level of V-ATPase H gene in the gills of S. constricta was significantly higher than that in the other 6 tissues including siphon, mantle, kidney, digestive gland, flabs, and foot (P < 0.01). Under salinity stress, the expression of V-ATPase H gene in the gills of each experimental group continued to rise, peaking at 24 h, which was significantly higher than that of the control group (P < 0.05). The experimental results show that the V-ATPase H gene plays a role in maintaining the balance between its own serum osmotic pressure and the external osmotic pressure in low-salt and high-salt environments during salinity adaptation of S. constricta.
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