Functional analysis of LHCSR1, a protein catalyzing NPQ in mosses, by heterologous expression in Arabidopsis thaliana

Photosynth Res. 2019 Dec;142(3):249-264. doi: 10.1007/s11120-019-00656-3. Epub 2019 Jul 3.


Non-photochemical quenching, NPQ, of chlorophyll fluorescence regulates the heat dissipation of chlorophyll excited states and determines the efficiency of the oxygenic photosynthetic systems. NPQ is regulated by a pH-sensing protein, responding to the chloroplast lumen acidification induced by excess light, coupled to an actuator, a chlorophyll/xanthophyll subunit where quenching reactions are catalyzed. In plants, the sensor is PSBS, while the two pigment-binding proteins Lhcb4 (also known as CP29) and LHCII are the actuators. In algae and mosses, stress-related light-harvesting proteins (LHCSR) comprise both functions of sensor and actuator within a single subunit. Here, we report on expressing the lhcsr1 gene from the moss Physcomitrella patens into several Arabidopsis thaliana npq4 mutants lacking the pH sensing PSBS protein essential for NPQ activity. The heterologous protein LHCSR1 accumulates in thylakoids of A. thaliana and NPQ activity can be partially restored. Complementation of double mutants lacking, besides PSBS, specific xanthophylls, allowed analyzing chromophore requirement for LHCSR-dependent quenching activity. We show that the partial recovery of NPQ is mostly due to the lower levels of Zeaxanthin in A. thaliana in comparison to P. patens. Complemented npq2npq4 mutants, lacking besides PSBS, Zeaxanthin Epoxidase, showed an NPQ recovery of up to 70% in comparison to A. thaliana wild type. Furthermore, we show that Lutein is not essential for the folding nor for the quenching activity of LHCSR1. In short, we have developed a system to study the function of LHCSR proteins using heterologous expression in a variety of A. thaliana mutants.

Keywords: A. thaliana; Heterologous expression; LHCSR; NPQ; P. patens; Photoprotection.

MeSH terms

  • Arabidopsis / genetics
  • Arabidopsis / metabolism*
  • Bryopsida / genetics*
  • Gene Expression Regulation, Plant
  • Light
  • Light-Harvesting Protein Complexes / genetics
  • Light-Harvesting Protein Complexes / metabolism*
  • Mutation
  • Oxidoreductases / genetics
  • Oxidoreductases / metabolism
  • Photochemical Processes
  • Photosynthesis
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism*
  • Plants, Genetically Modified
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Thylakoids / genetics
  • Thylakoids / metabolism
  • Xanthophylls / metabolism
  • Zeaxanthins / metabolism


  • Light-Harvesting Protein Complexes
  • Plant Proteins
  • Recombinant Proteins
  • Xanthophylls
  • Zeaxanthins
  • Oxidoreductases
  • zeaxanthin epoxidase