基于蛋白组学分析杜氏盐藻对不同浓度CO2的响应

用气体混合仪设置不同的CO_2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CO_2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CO_2浓度的升高,杜氏盐藻的生理活性及光合活性提高;而CO_2浓度过高对盐藻生长呈抑制作用; 3%CO_2浓度最适于本实验杜氏盐藻藻株的生长。随着CO_2浓度的升高,胞外碳酸酐酶活性下降。低浓度的CO_2有利于β-胡萝卜素的积累,且光系统Ⅱ(PSⅡ)光合活性在CO_2浓度0.03%～3%范围内上升, CO_2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CO_2浓度的变化;而过高浓度的CO_2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。     

硝基芳烃类物质对盐藻的毒性试验及比较

通过国际经合组织(OECD)藻类生长抑制试验的国际标准方法,参照生活饮用水卫生标准(GB5749-85)设置毒物浓度进行试验,得到了2,4-二硝基苯胺、2,4-二硝基氯苯、间二硝基苯对盐藻(Dunaliella salina)的48h半数有效抑制浓度EC50值,2,4-二硝基苯胺的为0.018mmol/L,2,4-二硝基氯苯的为0.015mmol/L,间二硝基苯为0.027mol/L。其毒性大小为2,4-二硝基氯苯>2,4-二硝基苯胺>间二硝基苯。并探讨了其致变的原因。     

盐藻过氧化物酶的盐适应性及其与物质积累的关系

实验分析了不同浓度NaCl处理下,培养盐藻的过氧化物酶(POD)活性及其与细胞密度、β-胡萝卜素积累和蛋白质积累的关系.结果表明,盐藻过氧化物酶活性随盐度变化而改变:在适当盐度(60～90g/L)下,过氧化物酶活性很低;在较低盐度(30～60g/L)或较高盐度(90～150g/L)下,盐藻过氧化物酶活性均显著升高,说明盐藻过氧化物酶是一种盐度逆境适应酶.盐藻过氧化物酶活性与盐藻细胞密度及物质积累关系密切:盐藻过氧化物酶活性很低时,盐藻细胞密度大,同时β-胡萝卜素和蛋白质积累也多;随着盐藻过氧化物酶活性升高,盐藻细胞密度、β-胡萝卜素和蛋白质积累均逐渐降低,但盐藻过氧化物酶活性进一步升高时,盐藻蛋白质积累又有增加,可能在盐藻体内有逆境蛋白产生.     

小球藻、盐藻光学特性的初步试验研究

利用分光光度计和赤潮生物快速检测仪对盐藻、小球藻的光谱特性、显微图像和荧光图像作了对比研究。结果表明:盐藻和小球藻都在400~450 nm之间(对应胡罗卜素)和650~700 nm之间(对应叶绿素)有吸收峰,但盐藻在近红外波段(960 nm附近)还有一峰值。泥沙的吸光值随波长增加单调递减。盐藻的荧光图像呈现红色,当有泥沙存在时,盐藻的荧光会被泥沙散射,该现象对于通过荧光图从泥沙中分辨藻是不利的。     

杜氏盐藻磷酸烯醇式丙酮酸羧化酶基因的克隆和分析

为研究杜氏盐藻（Dunaliella salina）磷酸烯醇式丙酮酸羧化酶（phosphoenolpyruvate carboxylase,PEPC）基因的功能, 根据莱茵衣藻（Chlamydomonas reinhardtii）、拟南芥（Arabidopsis thaliana）、花生（Arachis hypogae...     

Effect of nitrate and phosphate on accumulation of β-carotene isomers inDunaliella salina

Dunaliella salina, a halotolerant unicellular green alga, can accumulate a large amount of β-carotene under certain environmental conditions. The isomers of β-carotene extracted fromD. salina cultured in medium with different nitrate and phosphate concentrations were analysed by HPLC with Alox-T alumina column. At least six isomers were found in different proportions depending on the culture media's nitrate and/or phosphate concentrations. Nitrate and/or phosphate deficiency was conducive to the accumulation of total cis isomers but not of all trans isomer. It is suggested that 1 mmol/L KNO₃ and 0.1 mmol/L KH₂ PO₄ are favourable for accumulation of total cis β-carotene. Contribution No. 2090 from the Institute of Oceanology, Chinese Academy of Sciences. This project was supported by the National Natural Science Foundation of China, Grant No. 38970587.     

The effects of enriched CO2 and enhanced UV-B radiation on ultra-structure of Dunaliella salina, singly and in combination

1 Introduction The amount of stratospheric ozone was being re duced due to anthropogenic emission of chlorofluoro carbons (CFCs) and nitrogen orides (Molina and Row land, 1974; Rowland, 1989; Yang et al., 1998; Xiao e al., 2005). This reduction of ozone r…     

盐藻的室温光抑制

实验用盐藻取自天津塘沽轻工业部制盐研究所,采用ＡＳＰ２培养液为培养基,研究了盐藻的室温光抑制现象,并探讨了盐藻的光保护反应机理,结果表明,盐藻对高辐照有很强的忍耐力,且光合能力的恢复也很强,氯霉素（ＣＡＰ）或二硫苏糖醇（ＤＴＴ）在高光下强烈地加深了对ＰＳＩＩ光化学效率（Ｆｖ／Ｆｍ）和光合量子效率（Φｉ）的抑制,但ＣＡＰ或ＤＴＴ单独作用并不抑制盐藻的光化学系统在低光下的可逆恢复;盐藻在ＣＡＰ＋ＤＴＴ     

Expression,purification, and subcellular localization of phospholipase C in Dunaliella salina

Plants possess effective mechanisms to respond quickly to the external environment. Rapid activation of phosphatidylinositol-specific phospholipase C (PLC) enzymes occurs after a stimulus. The PLC in Dunaliella salina plays important roles in growth and stress responses. However, the molecular basis of PLC action in D. salina remains little understood. To gain insight into the potential biological functions of this enzyme, we cloned a phospholipase C gene from D. salina in a previous study, named DsPLC (GenBank No. KF573428). Here, we present the prokaryotic expression, purification, and characterization of the DsPLC gene. The entire coding region of DsPLC was inserted into an expression vector pET32a, and the DsPLC gene was successfully expressed in Escherichia coli. The DsPLC protein was purified and identified using a polyclonal antibody and western blotting. Expressing DsPLC fused with a green fluorescent protein (GFP) in onion showed that DsPLC-GFP was localized to the intracellular membrane. Quantitative real-time PCR analysis revealed that the relative expression of the DsPLC gene was induced significantly by 3.0-mol/L NaCl at 4 h. Our results support the importance of PLC enzymes in plant defense signaling. This study provides a basis for further functional studies of the DsPLC gene and for additional analysis of the potential roles of PLC enzymes in response to abiotic stress.