引用本文:	黄智慧,商晓梅,薛宝贵,马爱军,王新安,杨志,曲江波,王宝义.大菱鲆(Scophthalmus maximus)低温胁迫耐受性能与体表蛋白组学研究.海洋与湖沼,2013,44(3):638-644.

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*大菱鲆(Scophthalmus maximus)低温胁迫耐受性能与体表蛋白组学研究*
黄智慧,商晓梅,薛宝贵,马爱军,王新安,杨志,曲江波,王宝义
1.中国水产科学研究院黄海水产研究所农业部海洋渔业资源可持续利用重点开放实验室青岛市海水鱼类种子工程与生物技术重点实验室;2.烟台市开发区天源水产有限公司;3.兴城龙运井盐水水产养殖有限责任公司

摘要:

采用急性低温胁迫实验方法, 通过对实验鱼的死亡率、死亡历时、摄食量以及呼吸频率等耐寒指标进行比较分析, 并由此分析结果判断幼鱼的低温临界温度为0℃, 在此温度基础上, 利用双向电泳技术, 对大菱鲆(Scophthalmus maximus)幼鱼进行体表蛋白水平上的耐受机理研究。根据低温胁迫组和常温对照组机体表皮蛋白组图谱差异进行比较分析。结果表明, 低温胁迫后蛋白发生了显著变化。从中挑选四个差异蛋白点进行肽指纹图谱(MALDI-TOF-MS)分析, 采用PMF技术和MASCOT、NCBI网站提供的检索工具进行鉴定, 结果显示, 低温胁迫组大菱鲆表皮MLC和Mimecan前体蛋白表达显著上调, 而typeⅡ角蛋白表达显著下调。大菱鲆对低温胁迫是一个复杂的网络反应, 涉及很多蛋白质的参与, 这些低温响应蛋白在大菱鲆对低温胁迫的抗性反应中起到重要的作用。

关键词: 大菱鲆低温胁迫耐受性能体表蛋白双向电泳系统

DOI：10.11693/hyhz201303015015

分类号:

基金项目:现代农业产业技术体系建设专项资金, CARS-50-G01 号

附件

THE RESISTANCE ABILITY AND EPIDERMIS PROTEOMICS ANALYSIS OF TURBOT SCOPHTHALMUS MAXIMUS UNDER LOW TEMPERATURE STRESS

HUANG Zhi-Hui^1,2, SHANG Xiao-Mei^1,2, XUE Bao-Gui^1,2, MA Ai-Jun^1,2, WANG Xin-An^1,2, YANG Zhi³, QU Jiang-Bo³, WANG Bao-Yi⁴

1.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences;2.Key Lab for Sustainable Utilization of Marine Fishery Resources, Ministry of Agriculture, Qingdao Key Laboratory for MarineFish Breeding and Biotechnology;3.Yantai Tianyuan Aquatic Limited Corporation;4.Xingcheng Longyun Aquatic Limited Corporation

Abstract:

In order to assess the cold hardy capacity in the turbot (Scophthalmus maximus), this paper carried out the experiment of acute low temperature stress. In the condition of indoor artificial cooling, analyzed the resistant index by calculated the mortality, death time, food intake and breathing rate at static. And determine the critical temperature for juvenile is 0℃. Based on the temperature, we carried out the research about the tolerance mechanism in protein level by the technique of two-dimensional gel electrophoresis (2-DE). The epidermis proteome of were analyzed through after acute low temperature stress at 0℃, and then, analyzed by ImageMaster 2D Software. The result showed that significant changes in the expression between the normal temperature sample and the low stress sample. And 4 points of proteins were picked out to analyze through MALDI-TOF-MS, and then searched in database. The result demonstrated that the expressions of myosin light chain (MLC) and Mimecan precursor have up-regulated remarkably, however, type II keratin subunit protein down-regulated remarkably. It concluded that the responses of turbot to low temperature stress were considerable complex, including involvements of many proteins. And all those proteins played great roles in responses to low temperature stress for turbot. When the dynamic equilibrium in vivo environment of S. maximus was broken by low temperature stress, various physiological reactions would be induced and even death if the changes were out of control.

Key words: Turbot Scophthalmus maximus Low temperature stress Resistant index Epidermis proteins Two-dimensional gel electrophoresis (2-DE)