The Mehler reaction site is the Phylloquinone within Photosystem I

ABSTRACTPhotosynthesis is a vital process, responsible for fixing carbon dioxide, and producing most of the organic matter on the planet. However, photosynthesis has some inherent limitations in utilizing solar energy. Up to a third of the energy absorbed is lost in the reduction of O2 to produce the superoxide radical (O2•−), which occurs principally within photosystem I (PSI) via the Mehler reaction. Strikingly, the precise location as well as the evolutionary role of the reaction have long been a matter of debate. For decades, O2 reduction was assumed to take place solely in the distal iron-sulfur clusters of PSI rather than within the two asymmetrical cofactor branches. Here we demonstrate that under high irradiance, O2 photoreduction by PSI takes place at the phylloquinone of one of the branches (the A-branch). This conclusion derives from the light dependency of the O2 photoreduction rate constant, and from the high rates of O2 photoreduction in PSI complexes lacking iron-sulfur clusters and in a mutant PSI, in which the lifetime of this phyllosemiquinone state is extended 100-fold. On these grounds, we suggest that the Mehler reaction serves as a release valve, functioning only when needed, under conditions where both the distal iron-sulfur clusters of PSI and the mobile ferredoxin pool are over reduced.SIGNIFICANCE STATEMENTPhotosynthesis is the process responsible for the oxygenation of the ancient anoxic atmosphere, and the transformation of inorganic carbon to most of the organic matter on Earth. However, it is less commonly appreciated that the appearance of oxygen in the atmosphere led to the unavoidable opposite process in which oxygen is consumed, thereby producing deleterious oxygen radicals such as the superoxide radical. For almost half a decade, the location of the main site of superoxide radical production in chloroplasts has been a matter of debate. We now provide conclusive evidence that it is located in the phylloquinones(s) within photosystem I.

Dissecting the interaction of photosynthetic electron transfer with mitochondrial signalling and hypoxic response in the Arabidopsis rcd1 mutant

The Arabidopsis mutant rcd1 is tolerant to methyl viologen (MV). MV enhances the Mehler reaction, i.e. electron transfer from Photosystem I (PSI) to O 2 , generating reactive oxygen species (ROS) in the chloroplast. To study the MV tolerance of rcd1 , we first addressed chloroplast thiol redox enzymes potentially implicated in ROS scavenging. NADPH-thioredoxin oxidoreductase type C (NTRC) was more reduced in rcd1 . NTRC contributed to the photosynthetic and metabolic phenotypes of rcd1 , but did not determine its MV tolerance. We next tested rcd1 for alterations in the Mehler reaction. In rcd1 , but not in the wild type, the PSI-to-MV electron transfer was abolished by hypoxic atmosphere. A characteristic feature of rcd1 is constitutive expression of mitochondrial dysfunction stimulon (MDS) genes that affect mitochondrial respiration. Similarly to rcd1 , in other MDS-overexpressing plants hypoxia also inhibited the PSI-to-MV electron transfer. One possible explanation is that the MDS gene products may affect the Mehler reaction by altering the availability of O 2 . In green tissues, this putative effect is masked by photosynthetic O 2 evolution. However, O 2 evolution was rapidly suppressed in MV-treated plants. Transcriptomic meta-analysis indicated that MDS gene expression is linked to hypoxic response not only under MV, but also in standard growth conditions. This article is part of the theme issue ‘Retrograde signalling from endosymbiotic organelles’.

Different strategies of acclimation of photosynthesis, electron transport and antioxidative activity in leaves of two cotton species to water deficit

To better understand the adaptation mechanisms of the photosynthetic apparatus of cotton plants to water deficit conditions, the influence of water deficit on photosynthesis, chlorophyll a fluorescence and the activities of antioxidant systems were determined simultaneously in Gossypium hirsutum L. cv. Xinluzao 45 (upland cotton) and Gossypium barbadense L. cv. Xinhai 21 (pima cotton). Water deficit decreased photosynthesis in both cotton species, but did not decrease chlorophyll content or induce any sustained photoinhibition in either cotton species. Water deficit increased ETR/4 − AG, where ETR/4 estimates the linear photosynthetic electron flux and AG is the gross rate of carbon assimilation. The increase in ETR/4 − AG, which represents an increase in photorespiration and alternative electron fluxes, was particularly pronounced in Xinluzao 45. In Xinluzao 45, water deficit increased the activities of antioxidative enzymes, as well as the contents of reactive oxygen species (ROS), which are related to the Mehler reaction. In contrast, moderate water deficit particularly increased non-photochemical quenching (NPQ) in Xinhai 21. Our results suggest that Xinluzao 45 relied on enhanced electron transport such as photorespiration and the Mehler reaction to dissipate excess light energy under mild and moderate water deficit. Xinhai 21 used enhanced photorespiration for light energy utilisation under mild water deficit but, when subjected to moderate water deficit, possessed a high capacity for dissipating excess light energy via heat dissipation.

Interplay between Flavodiiron Proteins and Photorespiration in Synechocystis sp. PCC 6803

Flavodiiron (Flv) proteins are involved in detoxification of O2 and NO in anaerobic bacteria and archaea. Cyanobacterial Flv proteins, on the contrary, function in oxygenic environment and possess an extra NAD(P)H:flavin oxidoreductase module. Synechocystis sp. PCC 6803 has four genes (sll1521, sll0219, sll0550, and sll0217) encoding Flv proteins (Flv1, Flv2, Flv3, and Flv4). Previous in vitro studies with recombinant Flv3 protein from Synechocystis provided evidence that it functions as a NAD(P)H:oxygen oxidoreductase, and subsequent in vivo studies with Synechocystis confirmed the role of Flv1 and Flv3 proteins in the Mehler reaction (photoreduction of O2 to H2O). Interestingly, homologous proteins to Flv1 and Flv3 can be found also in green algae, mosses, and Selaginella. Here, we addressed the function of Flv1 and Flv3 in Synechocystis using the Δflv1, Δflv3, and Δflv1/Δflv3 mutants and applying inorganic carbon (Ci)-deprivation conditions. We propose that only the Flv1/Flv3 heterodimer form is functional in the Mehler reaction in vivo. 18O2 labeling was used to discriminate between O2 evolution in photosynthetic water splitting and O2 consumption. In wild type, ∼20% of electrons originated from water was targeted to O2 under air level CO2 conditions but increased up to 60% in severe limitation of Ci. Gas exchange experiments with Δflv1, Δflv3, and Δflv1/Δflv3 mutants demonstrated that a considerable amount of electrons in these mutants is directed to photorespiration under Ci deprivation. This assumption is in line with increased transcript abundance of photorespiratory genes and accumulation of photorespiratory intermediates in the WT and to a higher extent in mutant cells under Ci deprivation.