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基于蛋白组学分析杜氏盐藻对不同浓度CO2的响应
蔡学花1,2, 顾文辉1,3,4, 李元翔1,2, 王广策1,3,4
1.中国科学院海洋研究所实验海洋生物重点实验室, 山东 青岛 266071;2.中国科学院大学, 北京 100049;3.青岛国家海洋实验室 海洋生物与技术功能实验室, 山东 青岛 266237;4.中国科学院 海洋大科学研究中心, 山东 青岛 266071
摘要:
用气体混合仪设置不同的CO2浓度处理组,测定了杜氏盐藻(Dunaliella salina)细胞密度、碳酸酐酶活性、甘油含量等生理指标,结合蛋白质组学分析方法,比较了不同CO2浓度下细胞内主要代谢路径蛋白表达的差异。结果表明:在一定范围内,随着CO2浓度的升高,杜氏盐藻的生理活性及光合活性提高;而CO2浓度过高对盐藻生长呈抑制作用; 3% CO2浓度最适于本实验杜氏盐藻藻株的生长。随着CO2浓度的升高,胞外碳酸酐酶活性下降。低浓度的CO2有利于β-胡萝卜素的积累,且光系统Ⅱ(PSⅡ)光合活性在CO2浓度0.03%~3%范围内上升,CO2浓度达9%时降低,与光合作用相关蛋白的表达趋势接近。上述结果说明杜氏盐藻可能通过调节光合作用中叶绿素等捕光色素的合成及相关蛋白的表达笼统,以响应CO2浓度的变化;而过高浓度的CO2可对细胞产生氧化损害,引起热激蛋白和超氧化物歧化酶等蛋白含量的上调以应对氧化胁迫。
关键词:  杜氏盐藻  CO2浓度  蛋白组学  光合作用  氧化胁迫
DOI:10.11759/hykx20180327001
分类号:Q939.99
基金项目:国家自然科学基金项目(41506188);科技部国际合作专项(2015DFG32160)
The physiological and metabolic response of Dunaliella salina to different concentrations of CO2 based on proteomics
CAI Xue-hua1,2, GU Wen-hui1,3,4, LI Yuan-xiang1,2, WANG Guang-ce1,3,4
1.Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China;2.College of Earth Sciences, University of Chinese Academy of Sciences, Beijing 100049, China;3.Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China;4.Center for Ocean Mega-Science, Chinese Academy of Sciences, Qingdao 266071, China
Abstract:
To study the effect of different concentration of CO2 on Dunaliella salina, a gas mixing system, GMS 150, was used to produce precise gas mixtures with N2 and CO2. We investigated the growth rate, carbonic anhydrase (CA enzyme) activity, and glycerol content under different concentration of CO2. Differentially-expressed proteins in D. salina exposed to different CO2 concentrations were analyzed. Our results showed that both the physiological and photosynthetic activity increased with increasing CO2 concentration, while higher concentrations suppressed this trend. The optimal concentration for cell growth was 3% CO2. Extracellular CA enzyme activity declined with increasing CO2 concentration. β-Carotene accumulated under lower CO2, while higher CO2 facilitated glycerol synthesis. The photosynthetic activity of PSⅡ increased under 0.03% to 3% CO2 and was inhibited under 9% CO2. The relative expression of photosynthetic proteins was consistent with this trend. The synthesis and proportion of light-harvesting pigment was possibly regulated by changes in CO2 concentration. Higher CO2 concentrations may induce cellular oxidative stress. To cope with these negative factors, heat shock proteins and superoxide dismutase were up-regulated.
Key words:  Dunaliella salina  CO2 concentration  proteomic analysis  photosynthesis  oxidative stress
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