We report the first direct decomposition of the fluorescence lifetime heterogeneity during multiphasic fluorescence induction in dark-adapted leaves by multi-frequency phase and modulation fluorometry (PMF). A very fast component, assigned to photosystem I (PSI), was found to be constant in lifetime and yield, whereas the two slow components, which are strongly affected by the closure of the reaction centers by light, were assigned to PSII. Based on a modified "reversible radical pair" kinetic model with three compartments, we showed that a loosely connected pigment complex, which is assumed to be the CP47 complex, plays a specific role with respect to the structure and function of the PSII: (i) it explains the heterogeneity of PSII fluorescence lifetime as a compartmentation of excitation energy in the antenna, (ii) it is the site of a conformational change in the first second of illumination, and (iii) it is involved in the mechanisms of nonphotochemical quenching (NPQ). On the basis of the multi-frequency PMF analysis, we reconciled two apparently antagonistic aspects of chlorophyll a fluorescence in vivo: it is heterogeneous with respect to the kinetic structure (several lifetime components) and homogeneous with respect to average quantities (quasi-linear mean tau-Phi relationship).

译文

我们报告了通过多频相位和调制荧光法 (PMF) 在暗适应叶片中多相荧光诱导过程中荧光寿命异质性的首次直接分解。发现分配给光系统I (PSI) 的非常快的组件在寿命和产量上是恒定的,而两个慢的组件则被分配给PSII,这两个慢的组件受到光关闭反应中心的强烈影响。基于具有三个区室的改进的 “可逆自由基对” 动力学模型,我们证明了松散连接的颜料复合物 (假定为CP47复合物) 在PSII的结构和功能方面起着特定的作用: (i) 它解释了PSII荧光寿命的异质性作为激发能量在天线中的分隔,(ii) 它是照明的第一秒中构象变化的部位,并且 (iii) 它参与了非光化学猝灭 (NPQ) 的机制。在多频PMF分析的基础上,我们调和了体内叶绿素a荧光的两个明显拮抗方面: 它在动力学结构 (几个寿命分量) 方面是异质的,而在平均量方面是均匀的 (准线性平均tau-Phi关系)。

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