Comparative study on energy partitioning in photosystem II of two <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> mutants with reduced non-photochemical quenching capacity

Lhcb1-2 and PsbS proteins of photosystem II (PSII) have important roles in photoprotective thermal energy dissipation of the absorbed excess light energy. The light responses of chlorophyll fluorescence parameters were analyzed to examine how the absence of Lhcb1-2 or PsbS proteins can modify the energy allocation patterns of absorbed light energy in PSII using an antisense construct of lhcb2 and a psbS deletion (npq4-1) mutant of Arabidopsis thaliana. Both mutants exhibit reduced Stern–Volmer non-photochemical chlorophyll fluorescence quenching (NPQ). Here, we have adopted an approach, presented by Hendrickson et al. (Photosynth Res 82:73–81, 2004), to gain a better insight into the mechanism of the NPQ in these mutants. We have found no significant differences in the quantum yields of photochemical energy conversion (Φ_PSII) between the mutants and the wild type. Nevertheless, as it was expected, the fraction of the energy, which is dissipated as heat via regulated pathways in PSII (Φ_NPQ) for both mutants, were reduced as compared to the wild type. In a complementary way, the extent of non-regulated non-photochemical energy loss in PSII (Φ_NO) for both mutants was significantly higher than that in the wild type. This reflects, together with the lower Φ_NPQ (or NPQ) values, suboptimal capacity of photoprotective reactions at higher light intensities.     

The relationship between maximum tolerated light intensity and photoprotective energy dissipation in the photosynthetic antenna: chloroplast gains and losses

The principle of quantifying the efficiency of protection of photosystem II (PSII) reaction centres against photoinhibition by non-photochemical energy dissipation (NPQ) has been recently introduced by Ruban & Murchie (2012 Biochim. Biophys. Acta 1817, 977–982 (doi:10.1016/j.bbabio.2012.03.026)). This is based upon the assessment of two key parameters: (i) the relationship between the PSII yield and NPQ, and (ii) the fraction of intact PSII reaction centres in the dark after illumination. In this paper, we have quantified the relationship between the amplitude of NPQ and the light intensity at which all PSII reaction centres remain intact for plants with different levels of PsbS protein, known to play a key role in the process. It was found that the same, nearly linear, relationship exists between the levels of the protective NPQ component (pNPQ) and the tolerated light intensity in all types of studied plants. This approach allowed for the quantification of the maximum tolerated light intensity, the light intensity at which all plant leaves become photoinhibited, the fraction of (most likely) unnecessary or ‘wasteful’ NPQ, and the fraction of photoinhibited PSII reaction centres under conditions of prolonged illumination by full sunlight. It was concluded that the governing factors in the photoprotection of PSII are the level and rate of protective pNPQ formation, which are often in discord with the amplitude of the conventional measure of photoprotection, the quickly reversible NPQ component, qE. Hence, we recommend pNPQ as a more informative and less ambiguous parameter than qE, as it reflects the effectiveness and limitations of the major photoprotective process of the photosynthetic membrane.     

Acclimation- and mutation-induced enhancement of PsbS levels affects the kinetics of non-photochemical quenching in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

The efficiency of photosystem II antenna complexes (LHCs) in higher plants must be regulated to avoid potentially damaging overexcitation of the reaction centre in excess light. Regulation is achieved via a feedback mechanism known as non-photochemical quenching (NPQ), triggered the proton gradient (ΔpH) causing heat dissipation within the LHC antenna. ΔpH causes protonation of the LHCs, the PsbS protein and triggers the enzymatic de-epoxidation of the xanthophyll, violaxanthin, to zeaxanthin. A key step in understanding the mechanism is to decipher whether PsbS and zeaxanthin cooperate to promote NPQ. To obtain clues about their respective functions we studied the effects of PsbS and zeaxanthin on the rates of NPQ formation and relaxation in wild-type Arabidopsis leaves and those overexpressing PsbS (L17) or lacking zeaxanthin (npq1). Overexpression of PsbS was found to increase the rate of NPQ formation, as previously reported for zeaxanthin. However, PsbS overexpression also increased the rate of NPQ relaxation, unlike zeaxanthin, which is known decrease the rate. The enhancement of PsbS levels in plants lacking zeaxanthin (npq1) by either acclimation to high light or crossing with L17 plants showed that the effect of PsbS was independent of zeaxanthin. PsbS levels also affected the kinetics of the 535 nm absorption change (ΔA535), which monitors the formation of the conformational state of the LHC antenna associated with NPQ, in an identical way. The antagonistic action of PsbS and zeaxanthin with respect to NPQ and ΔA535 relaxation kinetics suggests that the two molecules have distinct regulatory functions.     

Elevated ΔpH restores rapidly reversible photoprotective energy dissipation in <Emphasis Type="Italic">Arabidopsis</Emphasis> chloroplasts deficient in lutein and xanthophyll cycle activity

The xanthophylls of the light-harvesting complexes of photosystem II (LHCII), zeaxanthin, and lutein are thought to be essential for non-photochemical quenching (NPQ). NPQ is a process of photoprotective energy dissipation in photosystem II (PSII). The major rapidly reversible component of NPQ, qE, is activated by the transmembrane proton gradient, and involves the quenching of antenna chlorophyll excited states by the xanthophylls lutein and zeaxanthin. Using diaminodurene (DAD), a mediator of cyclic electron flow around photosystem I, to enhance ΔpH we demonstrate that qE can still be formed in the absence of lutein and light-induced formation of zeaxanthin in chloroplasts derived from the normally qE-deficient lut2npq1 mutant of Arabidopsis. The qE induced by high ΔpH in lut2npq1 chloroplasts quenched the level of fluorescence when all PSII reaction centers were in the open state (F _o state), protected PSII reaction centers from photoinhibition, was sensitive to the uncoupler nigericin, and was accompanied by absorption changes in the 410–565 nm region. Titrations show the ΔpH threshold for activation of qE in lut2npq1 chloroplasts lies outside the normal physiological range and is highly cooperative. Comparison of quenching in isolated trimeric (LHCII) and monomeric (CP26) light-harvesting complexes from lut2npq1 plants revealed a similarly shifted pH dependency compared with wild-type LHCII. The implications for the roles of lutein and zeaxanthin as direct quenchers of excitation energy are discussed. Furthermore, we argue that the control over the proton-antenna association constant, pK, occurs via influence of xanthophyll structure on the interconnected phenomena of light-harvesting antenna reorganization/aggregation and hydrophobicity.     

Interaction between avoidance of photon absorption,excess energy dissipation and zeaxanthin synthesis against photooxidative stress in Arabidopsis

Plants evolved photoprotective mechanisms in order to counteract the damaging effects of excess light in oxygenic environments. Among them, chloroplast avoidance and non‐photochemical quenching concur in reducing the concentration of chlorophyll excited states in the photosynthetic apparatus to avoid photooxidation. We evaluated their relative importance in regulating excitation pressure on photosystem II. To this aim, genotypes were constructed carrying mutations impairing the chloroplast avoidance response (phot2) as well as mutations affecting the biosynthesis of the photoprotective xanthophyll zeaxanthin (npq1) or the activation of non‐photochemical quenching (npq4), followed by evaluation of their photosensitivity in vivo. Suppression of avoidance response resulted in oxidative stress under excess light at low temperature, while removing either zeaxanthin or PsbS had a milder effect. The double mutants phot2 npq1 and phot2 npq4 showed the highest sensitivity to photooxidative stress, indicating that xanthophyll cycle and qE have additive effects over the avoidance response. The interactions between non‐photochemical quenching and avoidance responses were studied by analyzing the kinetics of fluorescence decay and recovery at different light intensities. phot2 fluorescence decay lacked a component, here named as qM. This kinetic component linearly correlated with the leaf transmittance changes due to chloroplast relocation induced by white light and was absent when red light was used as actinic source. On these basis we conclude that a decrease in leaf optical density affects the apparent non‐photochemical quenching (NPQ) rise kinetic. Thus, excess light‐induced fluorescence decrease is in part due to avoidance of photon absorption rather than to a genuine quenching process.     

The role of the PsbS protein in the protection of photosystems I and II against high light in Arabidopsis thaliana

The PsbS protein is recognised in higher plants as an important component in dissipating excess light energy via its regulation of non-photochemical quenching. We investigated photosynthetic responses in the arabidopsis npq4 mutant, which lacks PsbS, and in a mutant over-expressing PsbS (oePsbS). Growth under low light led to npq4 and wild-type plants being visibly indistinguishable, but induced a phenotype in oePsbS plants, which were smaller and had shorter flowering spikes. Here we report that chloroplasts from npq4 generated more singlet oxygen (¹O₂) than those from oePsbS. This accompanied a higher extent of photosystem II photoinhibition of leaves from npq4 plants. In contrast, oePsbS was more damaged by high light than npq4 and the wild-type at the level of photosystem I. The plastoquinone pool, as measured by thermoluminescence, was more oxidised in the oePsbS than in npq4, whilst the amount of photo-oxidisable P₇₀₀, as probed with actinic light or saturating flashes, was higher in oePsbS compared to wild-type and npq4. Taken together, this indicates that the level of PsbS has a regulatory role in cyclic electron flow. Overall, we show that under high light oePsbS plants were more protected from ¹O₂ at the level of photosystem II, whereas lack of cyclic electron flow rendered them susceptible to damage at photosystem I. Cyclic electron flow is concluded to be essential for protecting photosystem I from high light stress.     

Reversible photoinactivation of photosystem II during desiccation of barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L. cv. Albori) leaves in the light

We investigated the inherent protective mechanisms against stress and damage to photosystem II (PSII) in barley (Hordeum vulgare L. cv. Albori). Leaves were desiccated at 30% relative humidity, under either low or high light (photon flux densities of 100 or 300 μmol m^-2 s^-1, respectively). During the treatment period, relative water content dropped to 35 to 45%, depending on light intensity. However, the photochemical efficiency of PSII (Fv/Fm) decreased only about 10%. This relatively stable response was due to the rapid, reversible increase in Fm (maximum fluorescence) during 20 min of dark-adaptation. During desiccation in the light, however, PSII was photo-inactivated by non-photochemical quenching (NPQ), with the excess excitation energy absorbed by the chlorophyll being dissipated as heat energy. This decline in NPQ in the first 2 min of treatment was caused by a relaxation in the energy-dependent quenching during dark-adaptation, but could be delayed significantly by a phosphatase inhibitor, NaF. In addition, the relaxation of other NPQ components related to state transition and phosphorylation of thylakoid phosphoproteins were blocked by NaF.     

Mechanisms of inhibitory effects of polycyclic aromatic hydrocarbons in photosynthetic primary processes in pea leaves and thylakoid preparations

• Inhibitory effects of polycyclic aromatic hydrocarbons (PAHs) on plants were studied in pea leaves in order to elucidate the mechanisms of action of PAHs such as naphthalene (Naph) and phenanthrene (Phen) on activity of photosystem II (PSII).
• The changes in different Chl fluorescence parameters were calculated on the basis of Chl fluorescence induction curves. H₂O₂ content was measured in leaf homogenates with the luminol‐dependent chemiluminescence method.
• We demonstrated that following PAH treatment, total energy dissipation (DI₀/ABS) and amount of Q_B‐non‐reducing complexes of PSII significantly increased. Non‐photochemical quenching (NPQ) also increased, when weak oxidative stress after PAH application developed. In leaves, a two‐step increase in H₂O₂ was found with time of incubation in the presence of PAHs, which may be associated with damage to the lipid bilayer of the plasma membrane and then violation of lipid bilayer membranes of cell organelles.
• A hypothesis for the mode of action of PAHs is provided that involves the role of ROS, membrane permeability and associated functional changes in PSII.

     

缺铁对大豆叶片光合作用和光系统Ⅱ功能的影响

通过气体交换和叶绿素荧光测定研究了缺铁对大豆叶片碳同化和光系统Ⅱ的影响。缺铁条件下大豆光合速率(Pn)大幅下降;最大光化学效率(po)下降幅度较小;荧光诱导动力学曲线发生明显的变化,其中电子传递活性明显下降,K相(VK)相对荧光产量提高。缺铁大豆的天线转化效率(Fv'/Fm')、光化学猝灭系数(qP)和光系统Ⅱ实际光化学效率(ΦPSⅡ)降低,而非光化学猝灭(NPQ)则明显增加。此外,缺铁大豆的光后荧光上升增强。据此,认为铁缺乏伤害了光系统Ⅱ复合物供体侧和受体侧的电子传递;缺铁条件下光系统I环式电子传递的增强可能在维持激发能耗散和ATP供给方面起一定作用。     

PSII photochemistry, thermal energy dissipation, and the xanthophyll cycle in Kalanchoë daigremontiana exposed to a combination of water stress and high light

Kalanchoë daigremontiana, a CAM plant grown in a greenhouse, was subjected to severe water stress. The changes in photosystem II (PSII) photochemistry were investigated in water‐stressed leaves. To separate water stress effects from photoinhibition, water stress was imposed at low irradiance (daily peak PFD 150 μmol m^?2 s^?1). There were no significant changes in the maximal efficiency of PSII photochemistry (F_v/F_m), the traditional fluorescence induction kinetics (OIP) and the polyphasic fluorescence induction kinetics (OJIP), suggesting that water stress had no direct effects on the primary PSII photochemistry in dark‐adapted leaves. However, PSII photochemistry in light‐adapted leaves was modified in water‐stressed plants. This was shown by the decrease in the actual PSII efficiency (Φ_PSII), the efficiency of excitation energy capture by open PSII centres (F_v′/F_m′), and photochemical quenching (q_P), as well as a significant increase in non‐photochemical quenching (NPQ) in particular at high PFDs. In addition, photoinhibition and the xanthophyll cycle were investigated in water‐stressed leaves when exposed to 50% full sunlight and full sunlight. At midday, water stress induced a substantial decrease in F_v/F_m which was reversible. Such a decrease was greater at higher irradiance. Similar results were observed in Φ_PSII, q_P, and F_v′/F_m′. On the other hand, water stress induced a significant increase in NPQ and the level of zeaxanthin via the de‐epoxidation of violaxanthin and their increases were greater at higher irradiance. The results suggest that water stress led to increased susceptibility to photoinhibition which was attributed to a photoprotective process but not to a photodamage process. Such a photoprotection was associated with the enhanced formation of zeaxanthin via de‐epoxidation of violaxanthin. The results also suggest that thermal dissipation of excess energy associated with the xanthophyll cycle may be an important adaptive mechanism to help protect the photosynthetic apparatus from photoinhibitory damage for CAM plants normally growing in arid and semi‐arid areas where they are subjected to a combination of water stress and high light.     

Expression and functional analysis of two PsbS genes in bamboo (Phyllostachys edulis)

Higher plants have an array of photoprotection mechanisms alleviating the harmful effects of light. Non‐photochemical quenching (NPQ) is one of the photoprotective mechanisms, which dissipates the excess of light energy absorbed in the light‐harvesting complexes (LHCs) into thermal energy. The photosystem II subunit S (PsbS), a member of the LHC family thought to be present exclusively in higher plants, is supposed to activate NPQ through interactions with antenna proteins. However, the roles of PsbS in bamboo remain unclear. Here, two genes of bamboo (Phyllostachys edulis), PePsbS1 and PePsbS2, are investigated and functionally analyzed. PePsbS1 and PePsbS2 have a similar gene structure with three introns separated by two exons, which encode 269 and 268 amino acid residues, respectively. Tissue‐specific analysis showed that PePsbS1 and PePsbS2 are highly expressed in leaf blade. Besides, they are both upregulated in the leaf blade when plantlets are submitted to an increased and prolonged light intensity, suggesting that they are light‐induced. Western blot analysis indicated that the accumulation level of total PePsbSs is consistent with what obtained by quantitative real‐time polymerase chain reaction for PePsbS1 and PePsbS2. Transgenic Arabidopsis plants overexpressing PePsbS1 and PePsbS2 both displayed an enhanced photoprotection. Moreover, the expression of PePsbS1 and PePsbS2 could both rescue the NPQ of Arabidopsis npq4 mutant, indicating that the PsbSs are functionally conserved between monocots and dicots. These results indicated that both PePsbS1 and PePsbS2 could circumvent photoinhibition and enhance photoprotection, which are key factors for bamboo's adaptation to different light environment.     

Restoration of rapidly reversible photoprotective energy dissipation in the absence of PsbS protein by enhanced DeltapH

Variations in the light environment require higher plants to regulate the light harvesting process. Under high light a mechanism known as non-photochemical quenching (NPQ) is triggered to dissipate excess absorbed light energy within the photosystem II (PSII) antenna as heat, preventing photodamage to the reaction center. The major component of NPQ, known as qE, is rapidly reversible in the dark and dependent upon the transmembrane proton gradient (ΔpH), formed as a result of photosynthetic electron transport. Using diaminodurene and phenazine metasulfate, mediators of cyclic electron flow around photosystem I, to enhance ΔpH, it is demonstrated that rapidly reversible qE-type quenching can be observed in intact chloroplasts from Arabidopsis plants lacking the PsbS protein, previously believed to be indispensible for the process. The qE in chloroplasts lacking PsbS significantly quenched the level of fluorescence when all PSII reaction centers were in the open state (F(o) state), protected PSII reaction centers from photoinhibition, was modulated by zeaxanthin and was accompanied by the qE-typical absorption spectral changes, known as ΔA(535). Titrations of the ΔpH dependence of qE in the absence of PsbS reveal that this protein affects the cooperativity and sensitivity of the photoprotective process to protons. The roles of PsbS and zeaxanthin are discussed in light of their involvement in the control of the proton-antenna association constant, pK, via regulation of the interconnected phenomena of PSII antenna reorganization/aggregation and hydrophobicity.     

Ascorbate-deficient mutants of Arabidopsis grow in high light despite chronic photooxidative stress

Acclimation to changing environments, such as increases in light intensity, is necessary, especially for the survival of sedentary organisms like plants. To learn more about the importance of ascorbate in the acclimation of plants to high light (HL), vtc2, an ascorbate-deficient mutant of Arabidopsis, and the double mutants vtc2npq4 and vtc2npq1 were tested for growth in low light and HL and compared with the wild type. The vtc2 mutant has only 10% to 30% of wild-type levels of ascorbate, vtc2npq4 has lower ascorbate levels and lacks non-photochemical quenching of chlorophyll fluorescence (NPQ) because of the absence of the photosystem II protein PsbS, and vtc2npq1 is NPQ deficient and also lacks zeaxanthin in HL but has PsbS. All three genotypes were able to grow in HL and had wild-type levels of Lhcb1, cytochrome f, PsaF, and 2-cysteine peroxiredoxin. However, the mutants had lower electron transport and oxygen evolution rates and lower quantum efficiency of PSII compared with the wild type, implying that they experienced chronic photooxidative stress. The mutants lacking NPQ in addition to ascorbate were only slightly more affected than vtc2. All three mutants had higher glutathione levels than the wild type in HL, suggesting a possible compensation for the lower ascorbate content. These results demonstrate the importance of ascorbate for the long-term acclimation of plants to HL.     

Low induction of non‐photochemical quenching and high photochemical efficiency in the annual desert plant Anastatica hierochuntica

Non‐photochemical quenching (NPQ) plays a major role in photoprotection. Anastatica hierochuntica is an annual desert plant found in hot deserts. We compared A. hierochuntica to three other different species: Arabidopsis thaliana, Eutrema salsugineum and Helianthus annuus, which have different NPQ and photosynthetic capacities. Anastatica hierochuntica plants had very different induction kinetics of NPQ and, to a lesser extent, of photosystem II electron transport rate (PSII ETR), in comparison to all other plants species in the experiments. The major components of the unusual photosynthetic and photoprotective response in A. hierochuntica were: (1) Low NPQ at the beginning of the light period, at various light intensities and CO₂ concentrations. The described low NPQ cannot be explained by low leaf absorbance or by low energy distribution to PSII, but was related to the de‐epoxidation state of xanthophylls. (2) Relatively high PSII ETR at various CO₂ concentrations in correlation with low NPQ. PSII ETR responded positively to the increase of CO₂ concentrations. At low CO₂ concentrations PSII ETR was mostly O₂ dependent. At moderate and high CO₂ concentrations the high PSII ETR in A. hierochuntica was accompanied by relatively high CO₂ assimilation rates. We suggest that A. hierochuntica have an uncommon NPQ and PSII ETR response. These responses in A. hierochuntica might represent an adaptation to the short growing season of an annual desert plant.     

On the origin of a slowly reversible fluorescence decay component in the Arabidopsis npq4 mutant

Over-excitation of photosynthetic apparatus causing photoinhibition is counteracted by non-photochemical quenching (NPQ) of chlorophyll fluorescence, dissipating excess absorbed energy into heat. The PsbS protein plays a key role in this process, thus making the PsbS-less npq4 mutant unable to carry out qE, the major and most rapid component of NPQ. It was proposed that npq4 does perform qE-type quenching, although at lower rate than WT Arabidopsis. Here, we investigated the kinetics of NPQ in PsbS-depleted mutants of Arabidopsis. We show that red light was less effective than white light in decreasing maximal fluorescence in npq4 mutants. Also, the kinetics of fluorescence dark recovery included a decay component, qM, exhibiting the same amplitude and half-life in both WT and npq4 mutants. This component was uncoupler-sensitive and unaffected by photosystem II repair or mitochondrial ATP synthesis inhibitors. Targeted reverse genetic analysis showed that traits affecting composition of the photosynthetic apparatus, carotenoid biosynthesis and state transitions did not affect qM. This was depleted in the npq4phot2 mutant which is impaired in chloroplast photorelocation, implying that fluorescence decay, previously described as a quenching component in npq4 is, in fact, the result of decreased photon absorption caused by chloroplast relocation rather than a change in the activity of quenching reactions.     

Effect of dichromate on photosystem II activity in xanthophyll-deficient mutants of Chlamydomonas reinhardtii

The photosystem II activity and energy dissipation was investigated when algal Chlamydomonas reinhardtii genotypes were exposed to dichromate toxicity effect. The exposure during 24 h to dichromate effect of two C. reinhardtii mutants having non-functional xanthophylls cycle, as npq1 zeaxanthin deficient and npq2 zeaxanthin accumulating, induced inhibition of PSII electron transport. After dichromate-induced toxicity, PSII functions of C. reinhardtii mutants were investigated under different light intensities. To determine dichromate toxicity and light intensity effect on PSII functional properties we investigated the change of energy dissipation via PSII electron transport, non-photochemical regulated and non-regulated energy dissipation according to Kramer et al. (Photosynth Res 79:209–218, 2004). We showed the dependency between dichromate toxicity and light-induced photoinhibition in algae deficient in xanthophyll cycle. When algal mutants missing xanthophylls cycle were exposed to dichromate toxicity and to high light intensity energy dissipation via non-regulated mechanism takes the most important pathway reaching the value of 80%. Therefore, the mutants npq1 and npq2 having non-functional xanthophylls cycle were more sensitive to dichromate toxic effects.     

Using chlorophyll fluorescence to assess the fraction of absorbed light allocated to thermal dissipation of excess excitation

In the present study we explored the possibility of assessing the allocation of photons absorbed by photosystem II (PSII) antennae to thermal energy dissipation and photosynthetic electron transport in leaves of several plant species under field conditions. Changes in chlorophyll fluorescence parameters were determined in situ over the course of an entire day in the field in sun-exposed leaves of two species with different maximal rates of photosynthesis, Helianthus annuus (sunflower) and Vinca major. Leaves of Vinca minor (periwinkle) growing in a deeply shaded location were also monitored. We propose using diurnal changes in the efficiency of open PSII centers (F′_v/F′_m) in these sun and shade leaves to (a) assess diurnal changes in the allocation of absorbed light to photochemistry and thermal energy dissipation and, furthermore, (b) make an estimate of changes in the rate of thermal energy dissipation, an analogous expression to the rate of photochemistry. The fraction of light absorbed in PSII antennae that is dissipated thermally (D) is proposed to be estimated from D = 1-F′_v/F′_m, in analogy to the widely used estimation of the fraction of light absorbed in PSII antennae (P) that is utilized in PSII photochemistry from P = F′_v/F′_m× q_P (where q_P is the coefficient for photochemical quenching; Genty, B., Briantais, J.-M. & Baker, N. R. 1989. Biochim. Biophys. Acta 990: 87-92). The rate of thermal dissipation is consequently given by D × PFD (photon flux density), again in analogy to the rate of photochemistry P × PFD, both assuming a matching behavior of photosystems I and II. Characterization of energy dissipation from the efficiency of open PSII centers allows an assessment from a single set of measurements at any time of day; this is particularly useful under field conditions where the fully relaxed reference values of variable or maximal fluorescence needed for the computation of nonphotochemical quenching may not be available. The usefulness of the assessment described above is compared with other currently used parameters to quantify nonphotochemical and photochemical chlorophyll fluorescence quenching.     

Molecular and global time-resolved analysis of a psbS gene dosage effect on pH- and xanthophyll cycle-dependent nonphotochemical quenching in photosystem II

Photosynthetic light harvesting in plants is regulated by a pH- and xanthophyll-dependent nonphotochemical quenching process (qE) that dissipates excess absorbed light energy and requires the psbS gene product. An Arabidopsis thaliana mutant, npq4-1, lacks qE because of a deletion of the psbS gene, yet it exhibits a semidominant phenotype. Here it is shown that the semidominance is due to a psbS gene dosage effect. Diploid Arabidopsis plants containing two psbS gene copies (wild-type), one psbS gene (npq4-1/NPQ4 heterozygote), and no psbS gene (npq4-1/npq4-1 homozygote) were compared. Heterozygous plants had 56% of the wild-type psbS mRNA level, 58% of the wild-type PsbS protein level, and 60% of the wild-type level of qE. Global analysis of the chlorophyll a fluorescence lifetime distributions revealed three components in wild-type and heterozygous plants, but only a single long lifetime component in npq4-1. The short lifetime distribution associated with qE was inhibited by more than 40% in heterozygous plants compared with the wild type. Thus, the extent of qE measured as either the fractional intensities of the PSII chlorophyll a fluorescence lifetime distributions or steady state intensities was stoichiometrically related to the amount of PsbS protein.