Crotty C. M., Tyrrell P. N., Espie G. S.
Department of Botany, Erindale College, University of Toronto, Mississauga, Ontario, Canada L5L 1C6.
Plant Physiol. 1994 Feb;104(2):785-791. doi: 10.1104/pp.104.2.785.
In the cyanobacterium Synechococcus UTEX 625, the yield of chlorophyll a fluorescence decreased in response to the transport-mediated accumulation of intracellular inorganic carbon (CO2 + HCO3- + CO32- = dissolved inorganic carbon [DIC]) and subsequently increased to a near-maximum level following photosynthetic depletion of the DIC pool. When DIC accumulation was mediated by the active Na+-dependent HCO3- transport system, the initial rate of fluorescence quenching was found to be highly correlated with the initial rate of H14CO3- transport (r = 0.96), and the extent of fluorescence quenching was correlated with the size of the internal DIC pool (r = 0.99). Na+-dependent HCO3- transport-mediated accumulation of DIC caused fluorescence quenching in either the presence or absence of the CO2 fixation inhibitor glycolaldehyde, indicating that quenching was not due simply to NADP+ reduction. The concentration of Na+ required to attain one-half the maximum rate of H14CO3- transport, at 20 [mu]M external HCO3-, declined from 9 to 1 mM as the external pH increased from 8 to 9.6. A similar pH dependency was observed when fluorescence quenching was used to determine the kinetic constants for HCO3- transport. In cells capable of Na+-dependent HCO3- transport, both the initial rate and extent of fluorescence quenching increased with increasing external HCO3-, saturating at about 150 [mu]M. In contrast Na+-independent HCO3- transport-mediated fluorescence quenching saturated at an HCO3- concentration of about 10 [mu]M. It was concluded that measurement of chlorophyll a fluorescence emission provided a convenient, but indirect, means of following Na+-dependent HCO3- transport and accumulation in Synechococcus.
在蓝藻聚球藻UTEX 625中,叶绿素a荧光产量会随着细胞内无机碳(CO₂ + HCO₃⁻ + CO₃²⁻ = 溶解无机碳[DIC])通过转运介导的积累而降低,随后在DIC库光合消耗后增加到接近最大值水平。当DIC积累由依赖Na⁺的主动HCO₃⁻转运系统介导时,发现荧光猝灭的初始速率与H¹⁴CO₃⁻转运的初始速率高度相关(r = 0.96),并且荧光猝灭的程度与内部DIC库的大小相关(r = 0.99)。依赖Na⁺的HCO₃⁻转运介导的DIC积累在有或没有CO₂固定抑制剂乙醇醛的情况下都会导致荧光猝灭,这表明猝灭不仅仅是由于NADP⁺还原。在外部HCO₃⁻为20 μM时,达到H¹⁴CO₃⁻转运最大速率一半所需的Na⁺浓度随着外部pH从8增加到9.6而从9 mM降至1 mM。当使用荧光猝灭来确定HCO₃⁻转运的动力学常数时,观察到类似的pH依赖性。在能够进行依赖Na⁺的HCO₃⁻转运的细胞中,荧光猝灭的初始速率和程度都随着外部HCO₃⁻浓度的增加而增加,在约150 μM时达到饱和。相比之下,不依赖Na⁺的HCO₃⁻转运介导的荧光猝灭在HCO₃⁻浓度约为10 μM时达到饱和。得出的结论是,叶绿素a荧光发射的测量提供了一种方便但间接的方法来跟踪聚球藻中依赖Na⁺的HCO₃⁻转运和积累。