The non-photochemical quenching protein LHCSR3 prevents oxygen-dependent photoinhibition in Chlamydomonas reinhardtii

Abstract Non-photochemical quenching (NPQ) helps dissipate surplus light energy, preventing formation of reactive oxygen species (ROS). In Chlamydomonas reinhardtii, the thylakoid membrane protein LHCSR3 is involved in pH-dependent (qE-type) NPQ, lacking in the npq4 mutant. Preventing PSII repair revealed that npq4 lost PSII activity faster than the wild type (WT) in elevated O2, while no difference between strains was observed in O2-depleted conditions. Low Fv/Fm values remained 1.5 h after moving cells out of high light, and this qH-type quenching was independent of LHCSR3 and not accompanied by losses of maximum PSII activity. Culturing cells in historic O2 atmospheres (30–35%) increased the qE of cells, due to increased LHCSR1 and PsbS levels, and LHCSR3 in the WT, showing that atmospheric O2 tensions regulate qE capacity. Colony growth of npq4 was severely restricted at elevated O2, and npq4 accumulated more reactive electrophile species (RES) than the WT, which could damage PSI. Levels of PsaA (PSI) were lower in npq4 grown at 35% O2, while PsbA (PSII) levels remained stable. We conclude that even at high O2 concentrations, the PSII repair cycle is sufficient to maintain net levels of PSII. However, LHCSR3 has an important function in protecting PSI against O2-mediated damage, such as via RES.

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The role of LHCBM1 in non-photochemical quenching in Chlamydomonas reinhardtii

10.1101/2022.01.13.476201 ◽

2022 ◽

Author(s):

Xin Liu ◽

Wojciech J Nawrocki ◽

Roberta Croce

Keyword(s):

Chlamydomonas Reinhardtii ◽

Protein Complexes ◽

Ph Sensor ◽

High Light ◽

Photochemical Quenching ◽

Knock Out ◽

Photosynthetic Organisms ◽

Pigment Protein Complexes ◽

Non Photochemical Quenching

Non-photochemical quenching (NPQ) is the process that protects photosynthetic organisms from photodamage by dissipating the energy absorbed in excess as heat. In the model green alga Chlamydomonas reinhardtii, NPQ was abolished in the knock-out mutants of the pigment-protein complexes LHCSR3 and LHCBM1. However, while LHCSR3 was shown to be a pH sensor and switching to a quenched conformation at low pH, the role of LHCBM1 in NPQ has not been elucidated yet. In this work, we combine biochemical and physiological measurements to study short-term high light acclimation of npq5, the mutant lacking LHCBM1. We show that while in low light in the absence of this complex, the antenna size of PSII is smaller than in its presence, this effect is marginal in high light, implying that a reduction of the antenna is not responsible for the low NPQ. We also show that the mutant expresses LHCSR3 at the WT level in high light, indicating that the absence of this complex is also not the reason. Finally, NPQ remains low in the mutant even when the pH is artificially lowered to values that can switch LHCSR3 to the quenched conformation. It is concluded that both LHCSR3 and LHCBM1 need to be present for the induction of NPQ and that LHCBM1 is the interacting partner of LHCSR3. This interaction can either enhance the quenching capacity of LHCSR3 or connect this complex with the PSII supercomplex.

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Ascorbate deficiency does not limit non-photochemical quenching in Chlamydomonas reinhardtii

10.1101/813766 ◽

2019 ◽

Author(s):

André Vidal-Meireles ◽

Dávid Tóth ◽

László Kovács ◽

Juliane Neupert ◽

Szilvia Z. Tóth

Keyword(s):

Chlamydomonas Reinhardtii ◽

Vascular Plants ◽

Regulation Of Gene Expression ◽

Growth Conditions ◽

Photochemical Quenching ◽

Wild Type ◽

Knockout Mutant ◽

Dependent Component ◽

Non Photochemical Quenching ◽

Energy Dependent

SummaryAscorbate (vitamin C) plays essential roles in development, signaling, hormone biosynthesis, regulation of gene expression, stress resistance and photoprotection. In vascular plants, violaxanthin de-epoxidase (VDE) requires ascorbate (Asc) as reductant, thereby it is required for the energy-dependent component of non-photochemical quenching (NPQ). In order to assess the role of Asc in NPQ in green algae, which are known to contain low amounts of Asc, we searched for an insertional Chlamydomonas reinhardtii mutant affected in the VTC2 gene, essential for Asc biosynthesis. The Crvtc2-1 knockout mutant was viable and, depending on the growth conditions, it contained 10 to 20% Asc relative to its wild type. When Chlamydomonas was grown photomixotrophically at moderate light, the zeaxanthin-dependent component of NPQ emerged upon strong red illumination both in the Crvtc2-1 mutant and in its wild type. Deepoxidation was unaffected by Asc deficiency, demonstrating that the Chlorophycean VDE found in Chlamydomonas does not require Asc as a reductant. The rapidly induced, energy-dependent NPQ component, characteristic of photoautotrophic Chlamydomonas cultures grown at high light, was not limited by Asc deficiency either. On the other hand, a reactive oxygen species-induced photoinhibitory NPQ component was greatly enhanced upon Asc deficiency, both under photomixotrophic and photoautotrophic conditions. These results demonstrate that Asc has distinct roles in NPQ formation in Chlamydomonas than in vascular plants.One-sentence summaryIn Chlamydomonas -in contrast to seed plants-, ascorbate is not required for violaxanthin deepoxidation and energy-dependent non-photochemical quenching but it mitigates photoinhibitory quenching.

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Non-photochemical quenching-dependent acclimation and thylakoid organization of Chlamydomonas reinhardtii to high light stress

Photosynthesis Research ◽

10.1007/s11120-018-0551-7 ◽

2018 ◽

Vol 139 (1-3) ◽

pp. 387-400 ◽

Cited By ~ 5

Author(s):

Srilatha Nama ◽

Sai Kiran Madireddi ◽

Ranay Mohan Yadav ◽

Rajagopal Subramanyam

Keyword(s):

Chlamydomonas Reinhardtii ◽

Light Stress ◽

High Light ◽

Photochemical Quenching ◽

High Light Stress ◽

Non Photochemical Quenching

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PsaK2 Subunit in Photosystem I Is Involved in State Transition under High Light Condition in the Cyanobacterium Synechocystis sp. PCC 6803

Journal of Biological Chemistry ◽

10.1074/jbc.m500369200 ◽

2005 ◽

Vol 280 (23) ◽

pp. 22191-22197 ◽

Cited By ~ 33

Author(s):

Tamaki Fujimori ◽

Yukako Hihara ◽

Kintake Sonoike

Keyword(s):

Energy Transfer ◽

Photosystem I ◽

State Transition ◽

Light Condition ◽

High Light ◽

Photochemical Quenching ◽

Wild Type ◽

Low Light ◽

High Light Condition ◽

Non Photochemical Quenching

To avoid the photodamage, cyanobacteria regulate the distribution of light energy absorbed by phycobilisome antenna either to photosystem II or to photosystem I (PSI) upon high light acclimation by the process so-called state transition. We found that an alternative PSI subunit, PsaK2 (sll0629 gene product), is involved in this process in the cyanobacterium Synechocystis sp. PCC 6803. An examination of the subunit composition of the purified PSI reaction center complexes revealed that PsaK2 subunit was absent in the PSI complexes under low light condition, but was incorporated into the complexes during acclimation to high light. The growth of the psaK2 mutant on solid medium was inhibited under high light condition. We determined the photosynthetic characteristics of the wild type strain and the two mutants, the psaK1 (ssr0390) mutant and the psaK2 mutant, using pulse amplitude modulation fluorometer. Non-photochemical quenching, which reflects the energy transfer from phycobilisome to PSI in cyanobacteria, was higher in high light grown cells than in low light grown cells, both in the wild type and the psaK1 mutant. However, this change of non-photochemical quenching during acclimation to high light was not observed in the psaK2 mutant. Thus, PsaK2 subunit is involved in the energy transfer from phycobilisome to PSI under high light condition. The role of PsaK2 in state transition under high light condition was also confirmed by chlorophyll fluorescence emission spectra determined at 77 K. The results suggest that PsaK2-dependent state transition is essential for the growth of this cyanobacterium under high light condition.

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Involvement of Lhcb6 and Lhcb5 in Photosynthesis Regulation in Physcomitrella patens Response to Abiotic Stress

International Journal of Molecular Sciences ◽

10.3390/ijms20153665 ◽

2019 ◽

Vol 20 (15) ◽

pp. 3665 ◽

Cited By ~ 3

Author(s):

Xingji Peng ◽

Xingguang Deng ◽

Xiaoya Tang ◽

Tinghong Tan ◽

Dawei Zhang ◽

...

Keyword(s):

Abiotic Stress ◽

Physcomitrella Patens ◽

Light Harvesting ◽

Light Stress ◽

High Light ◽

Photochemical Quenching ◽

Wild Type ◽

Knock Out ◽

Non Photochemical Quenching ◽

Photosynthesis Regulation

There are a number of highly conserved photosystem II light-harvesting antenna proteins in moss whose functions are unclear. Here, we investigated the involvement of chlorophyll-binding proteins, Lhcb6 and Lhcb5, in light-harvesting and photosynthesis regulation in Physcomitrella patens. Lhcb6 or Lhcb5 knock-out resulted in a disordered thylakoid arrangement, a decrease in the number of grana membranes, and an increase in the number of starch granule. The absence of Lhcb6 or Lhcb5 did not noticeably alter the electron transport rates. However, the non-photochemical quenching activity in the lhcb5 mutant was dramatically reduced when compared to wild-type or lhcb6 plants under abiotic stress. Lhcb5 plants were more sensitive to photo-inhibition, while lhcb6 plants showed little difference compared to the wild-type plants under high-light stress. Moreover, both mutants showed a growth malformation phenotype with reduced chlorophyll content in the gametophyte. These results suggested that Lhcb6 or Lhcb5 played a unique role in plant development, thylakoid organization, and photoprotection of PSII in Physcomitrella, especially when exposed to high light or osmotic environments.

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Photosystem II Subunit PsbS Is Involved in the Induction of LHCSR Protein-dependent Energy Dissipation in Chlamydomonas reinhardtii

Journal of Biological Chemistry ◽

10.1074/jbc.m116.737312 ◽

2016 ◽

Vol 291 (33) ◽

pp. 17478-17487 ◽

Cited By ~ 55

Author(s):

Viviana Correa-Galvis ◽

Petra Redekop ◽

Katharine Guan ◽

Annika Griess ◽

Thuy B. Truong ◽

...

Keyword(s):

Photosystem Ii ◽

Chlamydomonas Reinhardtii ◽

Conformational Changes ◽

Light Exposure ◽

Complex Stress ◽

High Light ◽

Photochemical Quenching ◽

Light Illumination ◽

Non Photochemical Quenching ◽

Psbs Protein

Non-photochemical quenching of excess excitation energy is an important photoprotective mechanism in photosynthetic organisms. In Arabidopsis thaliana, a high quenching capacity is constitutively present and depends on the PsbS protein. In the green alga Chlamydomonas reinhardtii, non-photochemical quenching becomes activated upon high light acclimation and requires the accumulation of light harvesting complex stress-related (LHCSR) proteins. Expression of the PsbS protein in C. reinhardtii has not been reported yet. Here, we show that PsbS is a light-induced protein in C. reinhardtii, whose accumulation under high light is further controlled by CO2 availability. PsbS accumulated after several hours of high light illumination at low CO2. At high CO2, however, PsbS was only transiently expressed under high light and was degraded after 1 h of high light exposure. PsbS accumulation correlated with an enhanced non-photochemical quenching capacity in high light-acclimated cells grown at low CO2. However, PsbS could not compensate for the function of LHCSR in an LHCSR-deficient mutant. Knockdown of PsbS accumulation led to reduction of both non-photochemical quenching capacity and LHCSR3 accumulation. Our data suggest that PsbS is essential for the activation of non-photochemical quenching in C. reinhardtii, possibly by promoting conformational changes required for activation of LHCSR3-dependent quenching in the antenna of photosystem II.

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Molecular genetics of xanthophyll–dependent photoprotection in green algae and plants

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.2000.0700 ◽

2000 ◽

Vol 355 (1402) ◽

pp. 1385-1394 ◽

Cited By ~ 103

Author(s):

Irene Baroli ◽

Krishna K. Niyogi

Keyword(s):

Reactive Oxygen Species ◽

Photosynthetic Apparatus ◽

Reactive Oxygen ◽

High Light ◽

Multiple Roles ◽

Oxygenic Photosynthesis ◽

Photochemical Quenching ◽

Oxygen Species ◽

Non Photochemical Quenching ◽

Extragenic Suppressors

The involvement of excited and highly reactive intermediates in oxygenic photosynthesis inevitably results in the generation of reactive oxygen species. To protect the photosynthetic apparatus from oxidative damage, xanthophyll pigments are involved in the quenching of excited chlorophyll and reactive oxygen species, namely 1 Chl*, 3 Chl*, and 1 1O 2 *. Quenching of 1 Chl* results in harmless dissipation of excitation energy as heat and is measured as non–photochemical quenching (NPQ) of chlorophyll fluorescence. The multiple roles of xanthophylls in photoprotection are being addressed by characterizing mutants of Chlamydomonas reinhardtii and Arabidopsis thaliana . Analysis of Arabidopsis mutants that are defective in 1 Chl* quenching has shown that, in addition to specific xanthophylls, the psbS gene is necessary for NPQ. Double mutants of Chlamydomonas and Arabidopsis that are deficient in zeaxanthin, lutein and NPQ undergo photo–oxidative bleaching in high light. Extragenic suppressors of the Chlamydomonas npq1 lor1 double mutant identify new mutations that restore varying levels of zeaxanthin accumulation and allow survival in high light.

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Light Absorption and Partitioning in Response to Nitrogen in Apple Leaves

HortScience ◽

10.21273/hortsci.33.3.541a ◽

1998 ◽

Vol 33 (3) ◽

pp. 541a-541

Author(s):

Lailiang Cheng ◽

Leslie H. Fuchigami ◽

Patrick J. Breen

Keyword(s):

High Light ◽

Thermal Dissipation ◽

Photochemical Quenching ◽

Light Conditions ◽

Psii Efficiency ◽

Fluorescence Measurements ◽

Free Radical Formation ◽

Highly Correlated ◽

Non Photochemical Quenching ◽

Leaf N

Bench-grafted Fuji/M26 apple trees were fertigated with different concentrations of nitrogen by using a modified Hoagland solution for 6 weeks, resulting in a range of leaf N from 1.0 to 4.3 g·m–2. Over this range, leaf absorptance increased curvilinearly from 75% to 92.5%. Under high light conditions (1500 (mol·m–2·s–1), the amount of absorbed light in excess of that required to saturate CO2 assimilation decreased with increasing leaf N. Chlorophyll fluorescence measurements revealed that the maximum photosystem II (PSII) efficiency of dark-adapted leaves was relatively constant over the leaf N range except for a slight drop at the lower end. As leaf N increased, non-photochemical quenching under high light declined and there was a corresponding increase in the efficiency with which the absorbed photons were delivered to open PSII centers. Photochemical quenching coefficient decreased significantly at the lower end of the leaf N range. Actual PSII efficiency increased curvilinearly with increasing leaf N, and was highly correlated with light-saturated CO2 assimilation. The fraction of absorbed light potentially used for free radical formation was estimated to be about 10% regardless of the leaf N status. It was concluded that increased thermal dissipation protected leaves from photo-oxidation as leaf N declined.

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Gradual Response of Cyanobacterial Thylakoids to Acute High-Light Stress—Importance of Carotenoid Accumulation

Cells ◽

10.3390/cells10081916 ◽

2021 ◽

Vol 10 (8) ◽

pp. 1916

Author(s):

Myriam Canonico ◽

Grzegorz Konert ◽

Aurélie Crepin ◽

Barbora Šedivá ◽

Radek Kaňa

Keyword(s):

Single Cell ◽

Thylakoid Membrane ◽

Intermediate Phase ◽

Light Stress ◽

Fast Response ◽

High Light ◽

Photochemical Quenching ◽

Second Phase ◽

Ros Scavenging ◽

Non Photochemical Quenching

Light plays an essential role in photosynthesis; however, its excess can cause damage to cellular components. Photosynthetic organisms thus developed a set of photoprotective mechanisms (e.g., non-photochemical quenching, photoinhibition) that can be studied by a classic biochemical and biophysical methods in cell suspension. Here, we combined these bulk methods with single-cell identification of microdomains in thylakoid membrane during high-light (HL) stress. We used Synechocystis sp. PCC 6803 cells with YFP tagged photosystem I. The single-cell data pointed to a three-phase response of cells to acute HL stress. We defined: (1) fast response phase (0–30 min), (2) intermediate phase (30–120 min), and (3) slow acclimation phase (120–360 min). During the first phase, cyanobacterial cells activated photoprotective mechanisms such as photoinhibition and non-photochemical quenching. Later on (during the second phase), we temporarily observed functional decoupling of phycobilisomes and sustained monomerization of photosystem II dimer. Simultaneously, cells also initiated accumulation of carotenoids, especially ɣ–carotene, the main precursor of all carotenoids. In the last phase, in addition to ɣ-carotene, we also observed accumulation of myxoxanthophyll and more even spatial distribution of photosystems and phycobilisomes between microdomains. We suggest that the overall carotenoid increase during HL stress could be involved either in the direct photoprotection (e.g., in ROS scavenging) and/or could play an additional role in maintaining optimal distribution of photosystems in thylakoid membrane to attain efficient photoprotection.

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