Department of Biotechnology, University of Verona, 37134 Verona, Italy.
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy.
Proc Natl Acad Sci U S A. 2019 Mar 5;116(10):4212-4217. doi: 10.1073/pnas.1809812116. Epub 2019 Feb 19.
Photosynthetic organisms prevent oxidative stress from light energy absorbed in excess through several photoprotective mechanisms. A major component is thermal dissipation of chlorophyll singlet excited states and is called nonphotochemical quenching (NPQ). NPQ is catalyzed in green algae by protein subunits called LHCSRs (Light Harvesting Complex Stress Related), homologous to the Light Harvesting Complexes (LHC), constituting the antenna system of both photosystem I (PSI) and PSII. We investigated the role of LHCSR1 and LHCSR3 in NPQ activation to verify whether these proteins are involved in thermal dissipation of PSI excitation energy, in addition to their well-known effect on PSII. To this aim, we measured the fluorescence emitted at 77 K by whole cells in a quenched or unquenched state, using green fluorescence protein as the internal standard. We show that NPQ activation by high light treatment in leads to energy quenching in both PSI and PSII antenna systems. By analyzing quenching properties of mutants affected on the expression of LHCSR1 or LHCSR3 gene products and/or state 1-state 2 transitions or zeaxanthin accumulation, namely, , , , , -complemented and , we showed that PSI undergoes NPQ through quenching of the associated LHCII antenna. This quenching event is fast-reversible on switching the light off, is mainly related to LHCSR3 activity, and is dependent on thylakoid luminal pH. Moreover, PSI quenching could also be observed in the absence of zeaxanthin or STT7 kinase activity.
光合作用生物通过几种光保护机制来防止过量吸收的光能产生氧化应激。一个主要组成部分是叶绿素单重激发态的热耗散,称为非光化学猝灭(NPQ)。NPQ 在绿藻中由称为 LHCSRs(与光收集复合物(LHC)同源的光收集复合物应激相关)的蛋白亚基催化,构成 PSI 和 PSII 的天线系统。我们研究了 LHCSR1 和 LHCSR3 在 NPQ 激活中的作用,以验证这些蛋白质是否除了对 PSII 的已知作用外,还参与 PSI 激发能的热耗散。为此,我们使用绿色荧光蛋白作为内标,测量了处于淬灭或未淬灭状态的整个细胞在 77 K 时发出的荧光。我们表明,高光处理在 中导致 PSI 和 PSII 天线系统中的能量猝灭。通过分析受 LHCSR1 或 LHCSR3 基因产物表达以及/或状态 1-状态 2 转变或玉米黄质积累影响的突变体的猝灭特性,即 、 、 、 、-互补 和 ,我们表明 PSI 通过与相关 LHCII 天线的猝灭来经历 NPQ。该猝灭事件在关闭光时快速可逆,主要与 LHCSR3 活性有关,并且依赖于类囊体腔内部的 pH 值。此外,即使没有玉米黄质或 STT7 激酶活性,也可以观察到 PSI 猝灭。