Rost Björn, Richter Klaus-Uwe, Riebesell Ulf, Hansen Per Juel
Alfred Wegener Institute for Polar and Marine Research, Am Handelshafen 12, 27515 Bremerhaven, Germany.
Plant Cell Environ. 2006 May;29(5):810-22. doi: 10.1111/j.1365-3040.2005.01450.x.
Carbon acquisition was investigated in three marine bloom-forming dinollagellates-Prorocentrum minimum, Heterocapsa triquetra and Ceratium lineatum. In vivo activities of extracellular and intracellular carbonic anhydrase (CA), photosynthetic O2 evolution, CO2 and HCO3- uptake rates were measured by membrane inlet mass spectrometry (MIMS) in cells acclimated to low pH (8.0) and high pH (8.5 or 9.1). A second approach used short-term 14C-disequilibrium incubations to estimate the carbon source utilized by the cells. All three species showed negligible extracellular CA (eCA) activity in cells acclimated to low pH and only slightly higher activity when acclimated to high pH. Intracellular CA (iCA) activity was present in all three species, but it increased only in P. minimum with increasing pH. Half-saturation concentrations (K1/2) for photosynthetic O2 evolution were low compared to ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) kinetics. Moreover, apparent affinities for inorganic carbon (Ci) increased with increasing pH in the acclimation, indicating the operation of an efficient CO2 concentration mechanism (CCM) in these dinoflagellates. Rates of CO2 uptake were comparably low and could not support the observed rates of photosynthesis. Consequently, rates of HCO3- uptake were high in the investigated species, contributing more than 80% of the photosynthetic carbon fixation. The affinity for HCO3- and maximum uptake rates increased under higher pH. The strong preference for HCO3- was also confirmed by the 14C-disequilibrium technique. Modes of carbon acquisition were consistent with the 13C-fractionation pattern observed and indicated a strong species-specific difference in leakage. These results suggest that photosynthesis in marine dinoflagellates is not limited by Ci even at high pH, which may occur during red tides in coastal waters.
对三种形成海洋水华的甲藻——微小原甲藻、三角异帽藻和线形角藻进行了碳获取的研究。在适应低pH值(8.0)和高pH值(8.5或9.1)的细胞中,通过膜进样质谱法(MIMS)测量了细胞外和细胞内碳酸酐酶(CA)的体内活性、光合放氧、二氧化碳和碳酸氢根的摄取速率。第二种方法是使用短期的14C不平衡培养来估计细胞利用的碳源。在适应低pH值的细胞中,所有这三个物种的细胞外CA(eCA)活性都可忽略不计,而在适应高pH值时,活性仅略有升高。所有这三个物种都存在细胞内CA(iCA)活性,但只有微小原甲藻的iCA活性随pH值升高而增加。与1,5-二磷酸核酮糖羧化酶/加氧酶(Rubisco)动力学相比,光合放氧的半饱和浓度(K1/2)较低。此外,在适应过程中,对无机碳(Ci)的表观亲和力随pH值升高而增加,表明这些甲藻中存在有效的二氧化碳浓缩机制(CCM)。二氧化碳摄取速率相对较低,无法支持观察到的光合速率。因此,在所研究的物种中,碳酸氢根的摄取速率很高,对光合碳固定的贡献超过80%。在较高pH值下,对碳酸氢根的亲和力和最大摄取速率增加。14C不平衡技术也证实了对碳酸氢根的强烈偏好。碳获取模式与观察到的13C分馏模式一致,并表明在渗漏方面存在强烈的物种特异性差异。这些结果表明,即使在高pH值下,海洋甲藻的光合作用也不受Ci的限制,而高pH值可能在沿海水域的赤潮期间出现。