Photoprotective Energy Dissipation Research Articles

Differential modulation of photosystem II photochemical efficiency in six C4 xero-halophytes.

Xero-halophytes are the salt-tolerant plants of dry habitats that adapt efficient strategies to endure extreme salt and water fluctuations. This study elucidated the adaptations related to PSII photochemistry, photoprotection, and photoinhibition in six C4 xero-halophytes (Atriplex stocksii , Haloxylon stocksii , Salsola imbricata, Suaeda fruticosa, Desmostachya bipinnata , and Saccharum griffithii ) grown in their native habitats. Chlorophyll a fluorescence quenching measurements suggested that S. imbricata and H. stocksii maintained efficient PSII photochemistry by downregulating heat dissipation and keeping a high fraction of open PSII centres that indicates plastoquinone (PQ) pool oxidation. Fluorescence induction kinetics revealed that S. imbricata demonstrated the highest performance index of PSII excitation to the reduction of end electron acceptors. S. fruticosa sustained photochemical efficiency through enhanced dissipation of excess energy and a low fraction of open PSII centres, indicating PQ reduced state. The large light-harvesting antenna size, deduced from the chlorophyll a /b ratio in S. fruticosa apparently led to the superior performance index of PSII excitation to the reduction of intersystem electron carriers. A. stocksii retained more open PSII centres with responsive non-photochemical quenching to safely dissipate excess energy. Despite maintaining the highest pigment contents and stoichiometry, A. stocksii remained lowest in both performance indices. The grass species D. bipinnata and S. griffithii kept fewer PSII centres open during photoinhibition, as evidenced by downregulation of PSII operating efficiency. The results provide insights into the differential modulation of PSII photochemical efficiency through dynamic control of photoprotective energy dissipation, PQ pool redox states, and photoinhibitory shutdown in these xero-halophytes.

EPR Spin-Trapping for Monitoring Temporal Dynamics of Singlet Oxygen during Photoprotection in Photosynthesis.

A central goal of photoprotective energy dissipation processes is the regulation of singlet oxygen (1O2*) and reactive oxygen species in the photosynthetic apparatus. Despite the involvement of 1O2* in photodamage and cell signaling, few studies directly correlate 1O2* formation to nonphotochemical quenching (NPQ) or lack thereof. Here, we combine spin-trapping electron paramagnetic resonance (EPR) and time-resolved fluorescence spectroscopies to track in real time the involvement of 1O2* during photoprotection in plant thylakoid membranes. The EPR spin-trapping method for detection of 1O2* was first optimized for photosensitization in dye-based chemical systems and then used to establish methods for monitoring the temporal dynamics of 1O2* in chlorophyll-containing photosynthetic membranes. We find that the apparent 1O2* concentration in membranes changes throughout a 1 h period of continuous illumination. During an initial response to high light intensity, the concentration of 1O2* decreased in parallel with a decrease in the chlorophyll fluorescence lifetime via NPQ. Treatment of membranes with nigericin, an uncoupler of the transmembrane proton gradient, delayed the activation of NPQ and the associated quenching of 1O2* during high light. Upon saturation of NPQ, the concentration of 1O2* increased in both untreated and nigericin-treated membranes, reflecting the utility of excess energy dissipation in mitigating photooxidative stress in the short term (i.e., the initial ∼10 min of high light).

Enhanced photochemical efficiency of PSII in Prosopis juliflora suggests contribution to invasion advantage over native C3 xero-halophytes under salt stress.

Chlorophyll a fluorescence parameters related to PSII photochemistry, photoprotection and photoinhibition were investigated in four C3 plant species growing in their natural habitat: Prosopis juliflora ; Abutilon indicum ; Salvadora persica ; and Phragmites karka . This study compared the light reaction responses of P. juliflora , an invasive species, with three native co-existing species, which adapt to varying water deficit and high salt stress. Chlorophyll a fluorescence quenching analyses revealed that P. juliflora had the highest photochemical quantum efficiency and yield, regulated by higher fraction of open reaction centres and reduced photoprotective energy dissipation without compromising the integrity of photosynthetic apparatus due to photoinhibition. Moreover, the elevated values of parameters obtained through polyphasic chlorophyll a fluorescence induction kinetics, which characterise the photochemistry of PSII and electron transport, highlighted the superior performance index of energy conservation in the transition from excitation to the reduction of intersystem electron carriers for P. juliflora compared to other species. Enhanced pigment contents and their stoichiometry in P. juliflora apparently contributed to upregulating fluxes and yields of energy absorbance, trapping and transport. This enhanced photochemistry, along with reduced non-photochemical processes, could explain the proclivity for invasion advantage in P. juliflora across diverse stress conditions.

Photosynthetic and transcriptome responses to fluctuating light in Arabidopsis thylakoid ion transport triple mutant.

Fluctuating light intensity challenges fluent photosynthetic electron transport in plants, inducing photoprotection while diminishing carbon assimilation and growth, and also influencing photosynthetic signaling for regulation of gene expression. Here, we employed in vivo chlorophyll-a fluorescence and P700 difference absorption measurements to demonstrate the enhancement of photoprotective energy dissipation of both photosystems in wild-type Arabidopsis thaliana after 6 h exposure to fluctuating light as compared with constant light conditions. This acclimation response to fluctuating light was hampered in a triple mutant lacking the thylakoid ion transport proteins KEA3, VCCN1, and CLCe, leading to photoinhibition of photosystem I. Transcriptome analysis revealed upregulation of genes involved in biotic stress and defense responses in both genotypes after exposure to fluctuating as compared with constant light, yet these responses were demonstrated to be largely upregulated in triple mutant already under constant light conditions compared with wild type. The current study illustrates the rapid acclimation of plants to fluctuating light, including photosynthetic, transcriptomic, and metabolic adjustments, and highlights the connection among thylakoid ion transport, photosynthetic energy balance, and cell signaling.

Unravelling the fluorescence kinetics of light-harvesting proteins with simulated measurements

The plant light-harvesting pigment-protein complex LHCII is the major antenna sub-unit of PSII and is generally (though not universally) accepted to play a role in photoprotective energy dissipation under high light conditions, a process known Non-Photochemical Quenching (NPQ). The underlying mechanisms of energy trapping and dissipation within LHCII are still debated. Various models have been proposed for the underlying molecular detail of NPQ, but they are often based on different interpretations of very similar transient absorption measurements of isolated complexes. Here we present a simulated measurement of the fluorescence decay kinetics of quenched LHCII aggregates to determine whether this relatively simple measurement can discriminate between different potential NPQ mechanisms. We simulate not just the underlying physics (excitation, energy migration, quenching and singlet-singlet annihilation) but also the signal detection and typical experimental data analysis. Comparing this to a selection of published fluorescence decay kinetics we find that: (1) Different proposed quenching mechanisms produce noticeably different fluorescence kinetics even at low (annihilation free) excitation density, though the degree of difference is dependent on pulse width. (2) Measured decay kinetics are consistent with most LHCII trimers becoming relatively slow excitation quenchers. A small sub-population of very fast quenchers produces kinetics which do not resemble any observed measurement. (3) It is necessary to consider at least two distinct quenching mechanisms in order to accurately reproduce experimental kinetics, supporting the idea that NPQ is not a simple binary switch.

The use of different LEDs wavelength and light intensities for in vitro proliferation of cherry rootstock: influence on photosynthesis and photomorphogenesis

Plant growth and development are strongly influenced by light quality conditions in their environment. In this study, in vitro shoot proliferation, photosynthetic pigments, leaf anatomy and photosystem II photochemistry of Gisela 6 cherry rootstock (Prunus cerasus x Prunus canescens) were investigated. The culture medium used was the MS (Murashige and Skoog) containing 4.4 μΜ 6-benzyladenine (BA), 0.05 μΜ α-naphthalene acetic acid (NAA) and 0.3 μM gibberellic acid (GA3) (30 days, 16 h photoperiod, 22 ± 2 °C). Initially, two different light wavelength absorption spectra: 400–700 nm provided by white fluorescent light lamps (mainly blue–green, BG at equal ratio 1:1; WFL-40) and, 430–690 nm provided by two Light Emitting Diodes (LED) types, differing in the spectral ratio of multiple light emitting wavelengths [LED2-40 (blue–red, BR) and LED3-40 (blue–green–yellow–orange–red, BGYOR)] were studied, under 40 μmol m−2 s−1 of light intensity. Following that, three LED intensities-20 (LED1-20), 40 (LED2-40), and 120 μmol m−2 s−1 (LED4-120) were investigated under BR spectrum. WFL-40 exhibited non-significant differences with LED2-40 (shoot number 4.25, proliferation rate 4.30, shoot height 0.91 cm). LED2-40 displayed higher fresh and dry shoot biomass, better PSII functionality, increased Chl a/b, Chl b and carotenoids content as well as decreased number of stressed explants. Leaf anatomy under LED2-40 and WFL-40 was similar. LED4-120 presented the highest rate of stressed explants, with their leaf anatomy being severely affected, as leaves had a reduced mesophyll cell density. In LED4-120, the proportion of the absorbed energy used in photochemistry (ΦPSII) was decreased whereas the increased quantum yield of non-regulated energy loss in PSII (ΦNO) could not be compensated by the increased photoprotective energy dissipation (ΦNPQ). Therefore, LED2-40 is proposed as the most beneficial for growing cherry rootstock Gisela 6 plantlets in vitro.

Wide variation of winter-induced sustained thermal energy dissipation in conifers: a common-garden study.

Low temperature in winter depresses rates of photosynthesis, which, in evergreen plants, can exacerbate imbalances between light absorption and photochemical light use. Damage that could result from increased excess light absorption is minimized by the conversion of excitation energy to heat in a process known as energy dissipation, which involves the de-epoxidized carotenoids of the xanthophyll cycle. Overwintering evergreens employ sustained forms of energy dissipation observable even after lengthy periods of dark acclimation. Whereas most studies of photoprotective energy dissipation examine one or a small number of species; here, we measured the levels of sustained thermal energy dissipation of seventy conifer taxa growing outdoors under common-garden conditions at the Red Butte Garden in Salt Lake City, Utah, U.S.A. (forty nine taxa were also sampled for needle pigment content). We observed an extremely wide range of wintertime engagement of sustained energy dissipation; the percentage decrease in dark-acclimated photosystem II quantum efficiency from summer to winter ranged from 6 to 95%. Of the many pigment-based parameters measured, the magnitude of the seasonal decrease in quantum efficiency was most closely associated with the seasonal increase in zeaxanthin content expressed on a total chlorophyll basis, which explained only slightly more than one-third of the variation. We did not find evidence for a consistent wintertime decrease in needle chlorophyll content. Thus, the prevailing mechanism for winter decreases in solar-induced fluorescence emitted by evergreen forests may be decreases in fluorescence quantum yield, and wintertime deployment of sustained energy dissipation likely underlies this effect.

Mechanisms shaping the synergism of zeaxanthin and PsbS in photoprotective energy dissipation in the photosynthetic apparatus of plants.

Safe operation of photosynthesis is vital to plants and is ensured by the activity of processes protecting chloroplasts against photo-damage. The harmless dissipation of excess excitation energy is considered to be the primary photoprotective mechanism and is most effective in the combined presence of PsbS protein and zeaxanthin, a xanthophyll accumulated in strong light as a result of the xanthophyll cycle. Here we address the problem of specific molecular mechanisms underlying the synergistic effect of zeaxanthin and PsbS. The experiments were conducted with Arabidopsis thaliana, using wild-type plants, mutants lacking PsbS (npq4), and mutants affected in the xanthophyll cycle (npq1), with the application of molecular spectroscopy and imaging techniques. The results lead to the conclusion that PsbS interferes with the formation of densely packed aggregates of thylakoid membrane proteins, thus allowing easy exchange and incorporation of xanthophyll cycle pigments into such structures. It was found that xanthophylls trapped within supramolecular structures, most likely in the interfacial protein region, determine their photophysical properties. The structures formed in the presence of violaxanthin are characterized by minimized dissipation of excitation energy. In contrast, the structures formed in the presence of zeaxanthin show enhanced excitation quenching, thus protecting the system against photo-damage.

Arctic sea ice algae differ markedly from phytoplankton in their ecophysiological characteristics

Photophysiological and biochemical characteristics were investigated in natural communities of Arctic sea ice algae and phytoplankton to understand their respective responses towards variable irradiance and nutrient regimes. This study revealed large differences in photosynthetic efficiency and capacity between the 2 types of algal assemblages. Sea ice algal assemblages clearly displayed increased photoprotective energy dissipation under the highest daily average irradiance levels (>8 µmol photons m-2 s-1). In contrast, phytoplankton assemblages were generally light-limited within the same irradiance ranges. Furthermore, phytoplankton assemblages exhibited more efficient carbon assimilation rates in the low irradiance range compared to sea ice algae, possibly explaining the ability of phytoplankton to generate substantial under-ice blooms. They were also able to readily adjust and increase their carbon production to higher irradiances. The Arctic is warming more rapidly than any other oceanic region on the planet, and as a consequence, irradiance levels experienced by microalgae are expected to increase due to declining ice thickness and snow cover, as well as enhanced stratification. The results of this study suggest that sea ice algae may have less capacity to adapt to the expected environmental changes compared to phytoplankton. We therefore anticipate a change in sea ice-based vs. pelagic primary production with respect to timing and quantity in a future Arctic. The clearly distinct responses of sea ice algae vs. phytoplankton need to be incorporated into model scenarios of current and future Arctic algal blooms and considered when predicting implications for the entire ecosystem and associated biogeochemical fluxes.

Effect of lhcsr gene dosage on oxidative stress and light use efficiency by Chlamydomonas reinhardtii cultures

Unicellular green algae, a promising source for renewable biofuels, produce lipid-rich biomass from light and CO2. Productivity in photo-bioreactors is affected by inhomogeneous light distribution from high cell pigment causing heat dissipation of light energy absorbed in excess and shading of the deep layers. Contrasting reports have been published on the relation between photoprotective energy dissipation and productivity. Here, we have re-investigated the relation between energy quenching (qE) activity, photodamage and light use efficiency by comparing WT and two Chlamydomonas reinhardtii strains differing for their complement in LHCSR proteins, which catalyse dissipation of excitation energy in excess (qE). Strains were analysed for ROS production, protein composition, rate of photodamage and productivity assessed under wide light and CO2 conditions.The strain lacking LHCSR1 and knocked down in LHCSR3, thus depleted in qE, produced O2 at significantly higher rate under high light, accompanied by enhanced singlet oxygen release and PSII photodamage. However, biomass productivity of WT was delayed in respect for mutant strains under intermittent light conditions only, implying that PSII activity was not the limiting factor under excess light. Contrary to previous proposals, domestication of Chlamydomonas for carbon assimilation rate in photo-bioreactors by down-regulation of photoprotective energy dissipation was ineffective in increasing algal biomass productivity.

Photoprotective energy dissipation is greater in the lower, not the upper, regions of a rice canopy: a 3D analysis.

High light intensities raise photosynthetic and plant growth rates but can cause damage to the photosynthetic machinery. The likelihood and severity of deleterious effects are minimised by a set of photoprotective mechanisms, one key process being the controlled dissipation of energy from chlorophyll within PSII known as non-photochemical quenching (NPQ). Although ubiquitous, the role of NPQ in plant productivity is important because it momentarily reduces the quantum efficiency of photosynthesis. Rice plants overexpressing and deficient in the gene encoding a central regulator of NPQ, the protein PsbS, were used to assess the effect of protective effectiveness of NPQ (pNPQ) at the canopy scale. Using a combination of three-dimensional reconstruction, modelling, chlorophyll fluorescence, and gas exchange, the influence of altered NPQ capacity on the distribution of pNPQ was explored. A higher phototolerance in the lower layers of a canopy was found, regardless of genotype, suggesting a mechanism for increased protection for leaves that experience relatively low light intensities interspersed with brief periods of high light. Relative to wild-type plants, psbS overexpressors have a reduced risk of photoinactivation and early growth advantage, demonstrating that manipulating photoprotective mechanisms can impact both subcellular mechanisms and whole-canopy function.

Changes in Photo-Protective Energy Dissipation of Photosystem II in Response to Beneficial Bacteria Consortium in Durum Wheat under Drought and Salinity Stresses

The present research aimed at evaluating the harmless dissipation of excess excitation energy by durum wheat (Triticum durum Desf.) leaves in response to the application of a bacterial consortium consisting of four plant growth-promoting bacteria (PGPB). Three pot experiments were carried out under non-stress, drought (at 40% field capacity), and salinity (150 mM NaCl) conditions. The results showed that drought and salinity affected photo-protective energy dissipation of photosystem II (PSII) increasing the rate of non-photochemical chlorophyll fluorescence quenching (NPQ (non-photochemical quenching) and qCN (complete non-photochemical quenching)), as well as decreasing the total quenching of chlorophyll fluorescence (qTQ), total quenching of variable chlorophyll fluorescence (qTV) and the ratio of the quantum yield of actual PSII photochemistry, in light-adapted state to the quantum yield of the constitutive non-regulatory NPQ (PQ rate). Our results also indicated that the PGPB inoculants can mitigate the adverse impacts of stresses on leaves, especially the saline one, in comparison with the non-fertilized (control) treatment, by increasing the fraction of light absorbed by the PSII antenna, PQ ratio, qTQ, and qTV. In the light of findings, our beneficial bacterial strains showed the potential in reducing reliance on traditional chemical fertilizers, in particular in saline soil, by improving the grain yield and regulating the amount of excitation energy.

Just the essentials: photoprotective energy dissipation pared-down.

This article comments on: Saccon F, Giovagnetti V, Shukla MK, Ruban AV. 2020. Rapid regulation of photosynthetic light harvesting in the absence of minor antenna and reaction centre complexes. Journal of Experimental Botany 71, 3626–3637.

A photosynthetic antenna complex foregoes unity carotenoid-to-bacteriochlorophyll energy transfer efficiency to ensure photoprotection

Carotenoids play a number of important roles in photosynthesis, primarily providing light-harvesting and photoprotective energy dissipation functions within pigment-protein complexes. The carbon-carbon double bond (C=C) conjugation length of carotenoids (N), generally between 9 and 15, determines the carotenoid-to-(bacterio)chlorophyll [(B)Chl] energy transfer efficiency. Here we purified and spectroscopically characterized light-harvesting complex 2 (LH2) from Rhodobacter sphaeroides containing the N = 7 carotenoid zeta (ζ)-carotene, not previously incorporated within a natural antenna complex. Transient absorption and time-resolved fluorescence show that, relative to the lifetime of the S1 state of ζ-carotene in solvent, the lifetime decreases ∼250-fold when ζ-carotene is incorporated within LH2, due to transfer of excitation energy to the B800 and B850 BChls a These measurements show that energy transfer proceeds with an efficiency of ∼100%, primarily via the S1 → Qx route because the S1 → S0 fluorescence emission of ζ-carotene overlaps almost perfectly with the Qx absorption band of the BChls. However, transient absorption measurements performed on microsecond timescales reveal that, unlike the native N ≥ 9 carotenoids normally utilized in light-harvesting complexes, ζ-carotene does not quench excited triplet states of BChl a, likely due to elevation of the ζ-carotene triplet energy state above that of BChl a These findings provide insights into the coevolution of photosynthetic pigments and pigment-protein complexes. We propose that the N ≥ 9 carotenoids found in light-harvesting antenna complexes represent a vital compromise that retains an acceptable level of energy transfer from carotenoids to (B)Chls while allowing acquisition of a new, essential function, namely, photoprotective quenching of harmful (B)Chl triplets.

A proteomic approach to seasonal adjustment in the enzyme complement of Korean fir (Abies koreana Wilson) needles

Korean fir is an endemic ornamental tree species facing population decline in Korea. To further understand the acclimatory adjustments it undergoes in response to seasonal extremes, we characterized some of the needle proteins that are upregulated during winter. Two-dimensional gel electrophoresis (2-DE), followed by MALDI TOF/TOF MS/MS and Mascot analyses, was used to visualize changes in protein profiles during acclimation to winter stress. From the 2-DE protein profiles of Korean fir needles, 226 protein spots were detected, many of which accumulated at higher levels during the winter. Among 17 proteins identified, 12 matched proteins associated with photosynthesis and with biotic and abiotic stresses, and eight were significantly upregulated under winter stress. Upregulated proteins included photosynthetic enzymes sedoheptulose-1,7-bisphosphatase and fructose bisphosphate aldolase of the Calvin–Benson cycle, four proteins related to oxidative stress tolerance, two proteins implicated in biotic defense, one heat-shock protein, and five unknown proteins. However, two other oxidative-stress-related proteins were present at high levels throughout the year, and a chitinase and the small subunit of ribulose-1,5-bisphosphate carboxylase showed no seasonal adjustments. Thus, Korean fir needles exhibited winter upregulation of some photosynthetic enzymes, coupled with increased photo protective thermal energy dissipation, and proteins related to abiotic and biotic stress resistance. Winter stress, which can include both low temperature and reduced water availability, in the subalpine region of Mount Halla led to an altered physiological equilibrium with increases in key Calvin–Benson cycle enzymes and increased enzymatic and non-enzymatic protection against oxidative stress.

The Plastid Lipocalin LCNP Is Required for Sustained Photoprotective Energy Dissipation in Arabidopsis

Light utilization is finely tuned in photosynthetic organisms to prevent cellular damage. The dissipation of excess absorbed light energy, a process termed nonphotochemical quenching (NPQ), plays an important role in photoprotection. Little is known about the sustained or slowly reversible form(s) of NPQ and whether they are photoprotective, in part due to the lack of mutants. The Arabidopsis thaliana suppressor of quenching1 (soq1) mutant exhibits enhanced sustained NPQ, which we term qH. To identify molecular players involved in qH, we screened for suppressors of soq1 and isolated mutants affecting either chlorophyllide a oxygenase or the chloroplastic lipocalin, now renamed plastid lipocalin (LCNP). Analysis of the mutants confirmed that qH is localized to the peripheral antenna (LHCII) of photosystem II and demonstrated that LCNP is required for qH, either directly (by forming NPQ sites) or indirectly (by modifying the LHCII membrane environment). qH operates under stress conditions such as cold and high light and is photoprotective, as it reduces lipid peroxidation levels. We propose that, under stress conditions, LCNP protects the thylakoid membrane by enabling sustained NPQ in LHCII, thereby preventing singlet oxygen stress.

Efficient modulation of photosynthetic apparatus confers desiccation tolerance in the resurrection plant Boea hygrometrica.

Boea hygrometrica (B. hygrometrica) can tolerate severe desiccation and resume photosynthetic activity rapidly upon water availability. However, little is known about the mechanisms by which B. hygrometrica adapts to dehydration and resumes competence upon rehydration. Here we determine how B. hygrometrica deals with oxidative stress, excessive excitation/electron pressures as well as photosynthetic apparatus modulation during dehydration/rehydration. By measuring ROS generation and scavenging efficiency, we found that B. hygrometrica possesses efficient strategies to maintain cellular redox homeostasis. Transmission electron microscopy (TEM) analysis revealed a remarkable alteration of chloroplast architecture and plastoglobules (PGs) accumulation during dehydration/rehydration. Pulse-amplitude modulated (PAM) chlorophyll fluorescence measurements, P700 redox assay as well as chlorophyll fluorescence emission spectra analysis on leaves of B. hygrometrica during dehydration/rehydration were also performed. Results showed that the photochemical activity of PSII as well as photoprotective energy dissipation in PSII undergo gradual inactivation/activation during dehydration/rehydration in B. hygrometrica; PSI activity is relatively induced upon water deficit, and dehydration leads to physical interaction between PSI and LHCII. Furthermore, blue-native polyacrylamide gel electrophoresis (BN-PAGE) and immunoblot analysis revealed that the protein abundance of light harvesting complexes decrease markedly along with internal water deficit to restrict light absorption and attenuate electron transfer, resulting in limited light excitation and repressed photosynthesis. In contrast, many thylakoid proteins remain at a basal level even after full dehydration. Taken together, our study demonstrated that efficient modulation of cellular redox homeostasis and photosynthetic activity confers desiccation tolerance in B. hygrometrica.

Remembering Navasard V. Karapetyan (1936-2015).

Navasard Vaganovich Karapetyan (September 6, 1936-March 6, 2015) began his scientific career at the Bach Institute of Biochemistry of the Russian Academy of Sciences, Moscow, and was associated with this institute for over 56 years. He worked in the area of biochemistry and biophysics of photosynthesis and was especially known for his studies on chlorophyll a fluorescence in higher plants and cyanobacteria, molecular organization of Photosystem I, photoprotective energy dissipation, and dynamics of energy migration in the two photosystems. We present here a brief biography and comments on the work of Navasard Karapetyan. We remember him as an enthusiastic person who had an unflagging curiosity, energy and profound sincere interest in many aspects of photosynthesis research.

Transient expression in Nicotiana benthamiana for rapid functional analysis of genes involved in non-photochemical quenching and carotenoid biosynthesis.

SummaryPlants must switch rapidly between light harvesting and photoprotection in response to environmental fluctuations in light intensity. This switch can lead to losses in absorbed energy usage, as photoprotective energy dissipation mechanisms can take minutes to hours to fully relax. One possible way to improve photosynthesis is to engineer these energy dissipation mechanisms (measured as non‐photochemical quenching of chlorophyll a fluorescence, NPQ) to induce and relax more quickly, resulting in smaller losses under dynamic light conditions. Previous studies aimed at understanding the enzymes involved in the regulation of NPQ have relied primarily on labor‐intensive and time‐consuming generation of stable transgenic lines and mutant populations – approaches limited to organisms amenable to genetic manipulation and mapping. To enable rapid functional testing of NPQ‐related genes from diverse organisms, we performed Agrobacterium tumefaciens‐mediated transient expression assays in Nicotiana benthamiana to test if NPQ kinetics could be modified in fully expanded leaves. By expressing Arabidopsis thaliana genes known to be involved in NPQ, we confirmed the viability of this method for studying dynamic photosynthetic processes. Subsequently, we used naturally occurring variation in photosystem II subunit S, a modulator of NPQ in plants, to explore how differences in amino acid sequence affect NPQ capacity and kinetics. Finally, we functionally characterized four predicted carotenoid biosynthesis genes from the marine algae Nannochloropsis oceanica and Thalassiosira pseudonana and examined the effect of their expression on NPQ in N. benthamiana. This method offers a powerful alternative to traditional gene characterization methods by providing a fast and easy platform for assessing gene function in planta.

Regulation and Levels of the Thylakoid K+/H+ Antiporter KEA3 Shape the Dynamic Response of Photosynthesis in Fluctuating Light.

Crop canopies create environments of highly fluctuating light intensities. In such environments, photoprotective mechanisms and their relaxation kinetics have been hypothesized to limit photosynthetic efficiency and therefore crop yield potential. Here, we show that overexpression of the Arabidopsis thylakoid K+/H+ antiporter KEA3 accelerates the relaxation of photoprotective energy-dependent quenching after transitions from high to low light in Arabidopsis and tobacco. This, in turn, enhances PSII quantum efficiency in both organisms, supporting that in wild-type plants, residual light energy quenching following a high to low light transition represents a limitation to photosynthetic efficiency in fluctuating light. This finding underscores the potential of accelerating quenching relaxation as a building block for improving photosynthetic efficiency in the field. Additionally, by overexpressing natural KEA3 variants with modification to the C-terminus, we show that KEA3 activity is regulated by a mechanism involving its lumen-localized C-terminus, which lowers KEA3 activity in high light. This regulatory mechanism fine-tunes the balance between photoprotective energy dissipation in high light and maximum quantum yield in low light, likely to be critical for efficient photosynthesis in fluctuating light conditions.