In nature, plants experience large fluctuations in light intensity and they need to balance the absorption and utilization of this energy accordingly. Non-photochemical quenching (NPQ) is a rapidly-switchable mechanism which protects plants from photodamage caused by high light exposure by dissipating the energy absorbed in excess as heat. It is triggered by the ΔpH across the thylakoid membrane and requires the presence of the protein PsbS and the xanthophyll zeaxanthin. However, the site and mechanism of the quencher(s) remain equivocal. Here, we constructed a mutant of Arabidopsis thaliana which lacks LHCII, the main antenna complexes of plants, to verify its contribution to NPQ. The mutant has normally stacked thylakoid membranes, displays no upregulation of other LHCs but shows a relative decrease in PSI which compensates for the decrease of the PSII antenna. The mutant exhibits a ~60% reduction in NPQ, while the remaining NPQ resembles that of the Chl bless mutant, which lacks all PSII peripheral antenna complexes. We thus report that PsbS-dependent NPQ mainly occurs within LHCII, but there is an additional quenching site within the PSII core. Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:

Photosynthesis is tightly regulated in order to withstand dynamic light environments. Under high light intensities, a mechanism known as non-photochemical quenching (NPQ) dissipates excess excitation energy, protecting the photosynthetic machinery from damage. An obstacle that lies in the way of understanding the molecular mechanism of NPQ is the large gap between in vitro and in vivo studies. On the one hand, the complexity of the photosynthetic membrane makes it challenging to obtain molecular information from in vivo experiments. On the other hand, a suitable in vitro system for the study of quenching is not available. Here we have developed a minimal NPQ system using proteoliposomes. With this, we demonstrate that the combination of low pH and PsbS is both necessary and sufficient to induce quenching in LHCII, the main antenna complex of plants. This proteoliposome system can be further exploited to gain more insight into how PsbS and other factors (e.g. zeaxanthin) influence the quenching mechanism observed in LHCII.

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Excess excitation energy in the light-harvesting antenna of Photosystem II (PSII) can cause irreversible damage to the photosynthetic apparatus. In periods of high light intensity, a feedback mechanism known as non-photochemical quenching (NPQ), induces the formation of quenchers which can safely dissipate excess excitation energy as heat. Although quenchers have been identified in more than one compartment of the PSII supercomplex, there is currently no quantitative description of how much NPQ is occurring at each of these locations. Here, we perform time-resolved fluorescence measurements on WT and antenna mutants lacking LHCII (NoL) and all peripheral antenna (Ch1 and Ch1lhcb5). By combining the results with those of steady-state fluorescence experiments we are able to estimate the intrinsic rate of NPQ for each plant and each PSII compartment. It is concluded that 60-70% of quenching occurs in LHCII, 15-20% in the minor antenna and 15-20% in the PSII core.