Formation of the Photosynthetic Electron Transport System during the Early Phase of Greening in Barley Leaves

The development of photochemical activity in isolated plastids during the early phase of greening of 5-day-old etiolated barley seedlings was studied and related to the appearance of chlorophyll-protein complexes. Photochemical activities of PSI (DCIPH₂ → MV) and PSII (H₂O → DCIP, DPC → DCIP) appeared at 1 and 1.5 hours after the onset of illumination, respectively. However, PSI + PSII activity (H₂O → MV, H₂O → NADP) appeared at 4 hours. The functional plastoquinone pool was noticed, at the latest, from 4 hours. Chloroplast preparations from seedlings of 1 h of greening showed O₂ uptake upon illumination in the absence of MV (−MV activity). This activity peaked at 2 hours of greening, then fell to zero by 6 hours. In contrast to the −MV activity, MV-Hill activity began to increase at 2 hours. Although PSI activity appeared at 1 hour, it failed to reduce ferredoxin until 2 hours. NADP began to be photoreduced at 4 hours in accordance with the appearance of the ferredoxin:NADP reductase activity. After formation of PSI and PSII, electron transport systems between them and between PSI and NADP developed in coordination with each other. Thus, the whole electron transport from water to NADP began to operate at 4 hours.     

Photosystem II activity of wild type <Emphasis Type="Italic">Synechocystis</Emphasis> PCC 6803 and its mutants with different plastoquinone pool redox states

To assess the role of redox state of photosystem II (PSII) acceptor side electron carriers in PSII photochemical activity, we studied sub-millisecond fluorescence kinetics of the wild type Synechocystis PCC 6803 and its mutants with natural variability in the redox state of the plastoquinone (PQ) pool. In cyanobacteria, dark adaptation tends to reduce PQ pool and induce a shift of the cyanobacterial photosynthetic apparatus to State 2, whereas illumination oxidizes PQ pool, leading to State 1 (Mullineaux, C. W., and Holzwarth, A. R. (1990) FEBS Lett., 260, 245-248). We show here that dark-adapted Ox^– mutant with naturally reduced PQ is characterized by slower Q_A^– reoxidation and O₂ evolution rates, as well as lower quantum yield of PSII primary photochemical reactions (F_v/F_m) as compared to the wild type and SDH–mutant, in which the PQ pool remains oxidized in the dark. These results indicate a large portion of photochemically inactive PSII reaction centers in the Ox^– mutant after dark adaptation. While light adaptation increases F_v/F_m in all tested strains, indicating PSII activation, by far the greatest increase in F_v/F_m and O₂ evolution rates is observed in the Ox^– mutant. Continuous illumination of Ox^– mutant cells with low-intensity blue light, that accelerates Q_A^– reoxidation, also increases F_v/F_m and PSII functional absorption cross-section (590 nm); this effect is almost absent in the wild type and SDH–mutant. We believe that these changes are caused by the reorganization of the photosynthetic apparatus during transition from State 2 to State 1. We propose that two processes affect the PSII activity during changes of light conditions: 1) reversible inactivation of PSII, which is associated with the reduction of electron carriers on the PSII acceptor side in the dark, and 2) PSII activation under low light related to the increase in functional absorption cross-section at 590 nm.     

Internal CO(2) Supply during Photosynthesis of Sun and Shade Grown CAM Plants in Relation to Photoinhibition

Leaves of Kalanchoë pinnata were exposed in the dark to air (allowing the fixation of CO₂ into malic acid) or 2% O₂, 0% CO₂ (preventing malic acid accumulation). They were then exposed to bright light in the presence or absence of external CO₂ and light dependent inhibition of photosynthetic properties assessed by changes in 77 K fluorescence from photosystem II (PSII), light response curves and quantum yields of O₂ exchange, rates of electron transport from H₂O through Q_B (secondary electron acceptor from the PSII reaction center) in isolated thylakoids, and numbers of functional PSII centers in intact leaf discs. Sun leaves of K. pinnata experienced greater photoinhibition when exposed to high light in the absence of CO₂ if malic acid accumulation had been prevented during the previous dark period. Shade leaves experienced a high degree of photoinhibition when exposed to high light regardless of whether malic acid had been allowed to accumulate in the previous dark period or not. Quantum yields were depressed to a greater degree than was 77 K fluorescence from PSII following photoinhibition.     

Organization of Electron Transport in Photosystem II of Spinach Chloroplasts According to Chelator Inhibition Sites

The organization of electron transport in photosystem II of spinach (Spinacia oleracea) chloroplasts was studied by means of various chelators and uncouplers. The partial reactions used included H₂O→methyl viologen, H₂O→silicomolybdic acid H₂O→ferricyanide, and H₂O→dimethylbenzoquinone. Three types of chelator inhibition were found (a) inhibition common to all pathways and presumably affecting the Mn or water oxidation site in photosystem II (salicylaldoxime, dithizone, acridine, 4,4,4-trifluoro-1-(2-thienyl)-1,1-butanedione, 4,4,4-trifluoro-0-(2-furyl)-1,3-butanedione; (b) strong inhibition of the H₂O→silicomolybdic acid pathway in presence of 3(3,4-dichlorophenyl)-1,1-dimethylurea by lipophilic chelators (bathocuproine, tertoctylcatechol) but stimulation by orthophenanthroline; and (c) 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone-insensitive dimethylbenzoquinone reduction inhibited by all phenanthrolines while ferricyanide reduction was remarkably stimulated by bathophenanthroline but inhibited by orthophenanthroline and bathocuproine. The action of lipophilic chelators on silicomolybdic acid reduction presumes the presence of a metallo protein in photosystem II. The differential action of bathophenanthroline on dimethylbenzoquinone and ferricyanide reduction indicated the possible existence of a metalloprotein in this pathway which is different from the site of orthophenanthroline inhibition.     

Photophosphorylation capacity of stable spheroplast preparations of anabaena

Spheroplasts from Anabaena 7119 (formerly designated Nostoc muscorum) were prepared in the presence of serum albumin in 0.5 molar sucrose. Electron transport and photophosphorylation were preserved (> 70% of the maximum rate for 1 week). The pH profile of electron transport and photophosphorylation in the reactions H₂O → NADP, H₂O → methyl viologen, and H₂O → ferricyanide shows that uncoupling by ammonia is small throughout and increases slightly with higher pH. ADP + Pi increased NADP reduction from H₂O by 2.5-fold. The ratios of ATP formed per electron pair transported ranged from 0.9 to 1.5. Effects of catalase and superoxide dismutase on the overall O₂ balance implicate pseudocyclic electron transport and phosphorylation. The quenching of 9-aminoacridine fluorescence indicates the formation of a Δ pH from 2 to 2.6 during illumination. This pH gradient is abolished by uncouplers; however, complete uncoupling is achieved only by 3-chlorocarbonyl cyanide phenylhydrazone or valinomycin + NH₄⁺. In the presence of NH₄⁺ alone, the membrane potential may act as the driving force for photophosphorylation.     

Desiccation tolerant lichens facilitate in vivo H/D isotope effect measurements in oxygenic photosynthesis

We have used the desiccation-tolerant lichen Flavoparmelia caperata, containing the green algal photobiont Trebouxia gelatinosa, to examine H/D isotope effects in Photosystem II in vivo. Artifact-free H/D isotope effects on both PSII primary charge separation and water oxidation yields were determined as a function of flash rate from chlorophyll-a variable fluorescence yields. Intact lichens could be reversibly dehydrated/re-hydrated with H₂O/D₂O repeatedly without loss of O₂ evolution, unlike all isolated PSII preparations. Above a threshold flash rate, PSII charge separation decreases sharply in both D₂O and H₂O, reflecting loss of excitation migration and capture by PSII. Changes in H/D coordinates further slow charge separation in D₂O (?23% at 120?Hz), attributed to reoxidation of the primary acceptor Q_A^?. At intermediate flash rates (5–50?Hz) D₂O decreases water oxidation efficiency (O₂ evolution) by ?2–5%. No significant isotopic difference is observed at slow flash rates (<5?Hz) where charge recombination dominates. Slower D₂O diffusion, changes in hydrogen bonding networks, and shifts in the pK_a's of ionizable residues may all contribute to these systematic variations of H/D isotope effects. Lichens' reversible desiccation tolerance allows highly reproducible H/D exchange kinetics in PSII reactions to be studied in vivo for the first time.     

Characterization of Photosystem II Activity and Heterogeneity during the Cell Cycle of the Green Alga Scenedesmus quadricauda

The photosynthetic activity of the green alga Scenedesmus quadricauda was investigated during synchronous growth in light/dark cycles. The rate of O₂ evolution increased 2-fold during the first 3 to 4 h of the light period, remained high for the next 3 to 4 h, and then declined during the last half of the light period. During cell division, which occurred at the beginning of the dark period, the ability of the cells to evolve O₂ was at a minimum. To determine if photosystem II (PSII) controls the photosynthetic capacity of the cells during the cell cycle we measured PSII activity and heterogeneity. Measurements of electron-transport activity revealed two populations of PSII, active centers that contribute to carbon reduction and inactive centers that do not. Measurements of PSII antenna sizes also revealed two populations, PSII_α and PSII_β, which differ from one another by their antenna size. During the early light period the photosynthetic capacity of the cells doubled, the O₂-evolving capacity of PSII was nearly constant, the proportion of PSII_β centers decreased to nearly zero, and the proportion of inactive PSII centers remained constant. During the period of minimum photosynthetic activity 30% of the PSII centers were insensitive to the inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea, which may be related to reorganization of the thylakoid membrane. We conclude from these results that PSII does not limit the photosynthetic activity of the cells during the first half of the light period. However, the decline in photosynthetic activity observed during the last half of the light period can be accounted for by limited PSII activity.     

Photosystem II Cyclic Heterogeneity and Photoactivation in the Diazotrophic, Unicellular Cyanobacterium Cyanothece Species ATCC 51142

The unicellular, diazotrophic cyanobacterium Cyanothece sp. ATCC 51142 demonstrated important modifications to photosystem II (PSII) centers when grown under light/dark N₂-fixing conditions. The properties of PSII were studied throughout the diurnal cycle using O₂-flash-yield and pulse-amplitude-modulated fluorescence techniques. Nonphotochemical quenching (q_N) of PSII increased during N₂ fixation and persisted after treatments known to induce transitions to state 1. The q_N was high in cells grown in the dark, and then disappeared progressively during the first 4 h of light growth. The photoactivation probability, ε, demonstrated interesting oscillations, with peaks near 3 h of darkness and 4 and 10 h of light. Experiments and calculations of the S-state distribution indicated that PSII displays a high level of heterogeneity, especially as the cells prepare for N₂ fixation. We conclude that the oxidizing side of PSII is strongly affected during the period before and after the peak of nitrogenase activity; changes include a lowered capacity for O₂ evolution, altered dark stability of PSII centers, and substantial changes in q_N.     

On the approaches applied in formulation of a kinetic model of photosystem II: Different approaches lead to different simulations of the chlorophyll a fluorescence transients

Chlorophyll a fluorescence rise (O-J-I-P transient) was in literature simulated using models describing reactions occurring solely in photosystem II (PSII) and plastoquinone (PQ) pool as well as using complex models which described, in addition to the above, also subsequent electron transport occurring beyond the PQ pool. However, there is no consistency in general approach how to formulate a kinetic model and how to describe particular reactions occurring even in PSII only. In this work, simple kinetic PSII models are considered always with the same electron carriers and same type of reactions but some reactions are approached in different ways: oxygen evolving complex is considered bound to PSII or “virtually” separated from PSII; exchange of doubly reduced secondary quinone PSII electron acceptor, Q_B, with PQ molecule from the PQ pool is described by one second order reaction or by two subsequent reactions; and all possible reactions or only those which follow in logical order are considered. By combining all these approaches, eight PSII models are formulated which are used for simulations of the chlorophyll a fluorescence transients. It is shown that the different approaches can lead to qualitatively different results. The approaches are compared with other models found elsewhere in the literature and therefore this work can help the readers to better understand the other models and their results.     

AN EXPERIMENTAL SEPARATION OF OXYGEN LIBERATION FROM CARBON DIOXIDE FIXATION IN PHOTOSYNTHESIS BY CHLORELLA

Using intact cells of Chlorella pyrenoidosa it is possible to obtain oxygen by the reduction of certain reducible materials other than carbon dioxide. Of these, benzaldehyde was studied in some detail. This reduction does not involve the production of carbon dioxide from the benzaldehyde. Stoichiometrical relationships as expressed by the following equation: 2C₆H₅CHO + 2H₂O → 2C₆H₅CH₂OH + O₂ are somewhat difficult to obtain because the benzaldehyde can disappear from the reaction mixtures by dark reactions. The technique is now available which permits detailed studies of the oxygen-liberating mechanisms in photosynthesis.     

Insensitivity of Water-Oxidation and Photosystem II Activity in Tomato to Chilling Temperatures

Chilling tomato plants (Lycopersicon esculentum Mill. cv. Rutgers and cv. Floramerica) in the dark resulted in a sizable inhibition in the rate of light- and CO₂-saturated photosynthesis. However, at low light intensity, the inhibition disappeared and the absolute quantum yield of CO₂ reduction was diminished only slightly. The quantum yield of photosystem II (PSII) electron flow was 18% lower when measured in chloroplasts isolated from chilled leaves than in chloroplasts isolated from unchilled leaves. Even though the maximum rate of PSII turnover in these chloroplasts was 12% lower subsequent to chilling, it was in all cases two or more times that required to support the light- and CO₂-saturated rate of photosynthesis measured in the attached leaf. The concentration of active PSII centers in chloroplasts isolated from leaves either before or after chilling was determined by measurement of the products of water oxidation from a series of saturating flashes short enough to turnover the electron transport carriers only a single time. There was no significant change in the concentration of active PSII centers due to dark chilling.

It was concluded that PSII activity and water oxidation capacity are not significantly impaired in tomato by chilling in the dark and therefore are not primary aspects of the inhibition of CO₂ reduction observed in attached leaves.

     

Interaction of exogenous quinones with membranes of higher plant chloroplasts: modulation of quinone capacities as photochemical and non-photochemical quenchers of energy in Photosystem II during light-dark transitions

Light modulation of the ability of three artificial quinones, 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB), 2,6-dichloro-p-benzoquinone (DCBQ), and tetramethyl-p-benzoquinone (duroquinone), to quench chlorophyll (Chl) fluorescence photochemically or non-photochemically was studied to simulate the functions of endogenous plastoquinones during the thermal phase of fast Chl fluorescence induction kinetics. DBMIB was found to suppress by severalfold the basal level of Chl fluorescence (F_o) and to markedly retard the light-induced rise of variable fluorescence (F_v). After irradiation with actinic light, Chl fluorescence rapidly dropped down to the level corresponding to F_o level in untreated thylakoids and then slowly declined to the initial level. DBMIB was found to be an efficient photochemical quencher of energy in Photosystem II (PSII) in the dark, but not after prolonged irradiation. Those events were owing to DBMIB reduction under light and its oxidation in the dark. At high concentrations, DCBQ exhibited quenching behaviours similar to those of DBMIB. In contrast, duroquinone demonstrated the ability to quench F_v at low concentration, while F_o was declined only at high concentrations of this artificial quinone. Unlike for DBMIB and DCBQ, quenched F_o level was attained rapidly after actinic light had been turned off in the presence of high duroquinone concentrations. That finding evidenced that the capacity of duroquinone to non-photochemically quench excitation energy in PSII was maintained during irradiation, which is likely owing to the rapid electron transfer from duroquinol to Photosystem I (PSI). It was suggested that DBMIB and DCBQ at high concentration, on the one hand, and duroquinone, on the other hand, mimic the properties of plastoquinones as photochemical and non-photochemical quenchers of energy in PSII under different conditions. The first model corresponds to the conditions under which the plastoquinone pool can be largely reduced (weak electron release from PSII to PSI compared to PSII-driven electron flow from water under strong light and weak PSI photochemical capacity because of inactive electron transport on its reducing side), while the second one mimics the behaviour of the plastoquinone pool when it cannot be filled up with electrons (weak or moderate light and high photochemical competence of PSI).     

Fermentative Metabolism of Hydrogen-evolving Chlamydomonas moewusii

The anaerobic metabolism of Chlamydomonas moewusii under both light (160 lux) and dark conditions has been examined using manometric and enzymic techniques. During anaerobiosis starch is broken down to glycerol, acetate, ethanol, CO₂, and H₂. The release of CO₂ and H₂ comes to an end when the starch pool is depleted.

There are only slight differences in the ratio of the end products of fermentation between light and dark metabolism. In the light, glycerol production is diminished and H₂ evolution is enhanced, whereas the production rate of all other end products generally does not change.

     

Studies on the photoactivation of the water-oxidizing enzyme : I. Processes limiting photoactivation in hydroxylamine-extracted leaf segments

In weak yet optimal light intensity, complete photoactivation of the water-oxidizing enzyme in NH₂OH-extracted wheat (Triticum aestivum, var Oasis) leaf segments could be obtained only after long dark preincubation. Photoactivation was not affected by ethylenediaminetetraacetate or inhibitors of photophosphorylation and protein synthesis, but was partially inhibited by a divalent cation ionophore. Complete photoactivation required ligation of ~4 Mn by the water oxidizing enzyme.

Without dark preincubation, photosystem II (PSII) was susceptible to weak light photoinhibition resulting in: (a) 50% maximum decrease in photooxidation of artificial electron donors by PSII: (b) increased times for the variable fluorescence rise (with 3-(3,4-dichlorophenyl)-1,1-dimethyl urea): (c) abolishment of photoactivation: and (d) the imposition of sensitivity to inhibitors of photophosphorylation and 70S but not 80S protein synthesis on subsequent light-dependent recovery from photoinhibition and recovery of O₂ evolution. Decrease in susceptibility to photoinhibition and increase in rates of photoactivation resulting from dark preincubations proved closely correlated. Neither protein synthesis nor increases in abundances of thylakoid Mn²⁺ and Ca²⁺ were required for escape from photoinhibition. However, photoactivation of the wateroxidizing enzyme in NH₂OH-extracted Chlamydomonas occurred in absence of dark preincubation and protein synthesis. Results are discussed in the context of disassembly/reassembly/resynthesis of specific PSII polypeptides.

     

Impact of PsbTc on forward and back electron flow, assembly, and phosphorylation patterns of photosystem II in tobacco

Photosystem II (PSII) of oxygen-evolving cyanobacteria, algae, and land plants mediates electron transfer from the Mn₄Ca cluster to the plastoquinone pool. It is a dimeric supramolecular complex comprising more than 30 subunits per monomer, of which 16 are bitopic or peripheral, low-molecular-weight components. Directed inactivation of the plastid gene encoding the low-molecular-weight peptide PsbTc in tobacco (Nicotiana tabacum) does not prevent photoautotrophic growth. Mutant plants appear normal green, and levels of PSII proteins are not affected. Yet, PSII-dependent electron transport, stability of PSII dimers, and assembly of PSII light-harvesting complexes (LHCII) are significantly impaired. PSII light sensitivity is moderately increased and recovery from photoinhibition is delayed, leading to faster D1 degradation in ΔpsbTc under high light. Thermoluminescence emission measurements revealed alterations of midpoint potentials of primary/secondary electron-accepting plastoquinone of PSII interaction. Only traces of CP43 and no D1/D2 proteins are phosphorylated, presumably due to structural changes of PSII in ΔpsbTc. In striking contrast to the wild type, LHCII in the mutant is phosphorylated in darkness, consistent with its association with PSI, indicating an increased pool of reduced plastoquinone in the dark. Finally, our data suggest that the secondary electron-accepting plastoquinone of PSII site, the properties of which are altered in ΔpsbTc, is required for oxidation of reduced plastoquinone in darkness in an oxygen-dependent manner. These data present novel aspects of plastoquinone redox regulation, chlororespiration, and redox control of LHCII phosphorylation.     

Damage and protection of the photosynthetic apparatus from UV-B radiation. I. Effect of ascorbate

In this work, the effect of the exogenously added ascorbate (Asc) against the UV-B inhibition of the photosystem II (PSII) functions in isolated pea thylakoid membranes was studied. The results reveal that Asc decreases the UV-B induced damage of the donor and the acceptor side of PSII during short treatment up to 60 min. The exogenous Asc exhibits a different UV-protective effect on PSII centers in grana and stroma lamellae, as the effect is more pronounced on the PSIIβ centers in comparison to PSIIα centers. Data also suggest that one of the possible protective roles of the Asc in photosynthetic membranes is the modification of the oxygen-evolving complex by influence on the initial S₀–S₁ state distribution in the dark.     

The Effect of Sulfur Re-Addition on H<Subscript>2</Subscript> Photoproduction by Sulfur-Deprived Green Algae

Sulfur deprivation of algal cultures selectively and partially inactivates photosystem II (PSII)-catalyzed O₂ evolution, induces anaerobiosis and hydrogenase expression, and results in sustained H₂ photoproduction for several days. We show that re-addition of limiting amounts of sulfate (1–10 μM final concentration) to the cultures during the H₂-production phase temporarily reactivates PSII photochemical and O₂-evolution activity and re-establishes higher rates of electron transport through the photosynthetic electron transport chain. The reactivation of PSII occurs by de novo D1 protein synthesis, but does not result in the re-establishment of aerobic conditions in the reactor, detectable by dissolved-O₂ sensors. However, concomitant H₂ photoproduction is inhibited, possibly due to excessive intra-cellular levels of photosynthetically-evolved O₂. The partial recovery of electron transport rates correlates with the re-oxidation of the plastoquinone (PQ) pool, as observed by pulse-amplitude modulated (PAM) and fluorescence-induction measurements. These results show that the presence of a more oxidized PQ pool releases some of the down-regulation of electron transport caused by the anaerobic conditions.     

Choline oxidation by intact spinach chloroplasts

Plants synthesize betaine by a two-step oxidation of choline (choline → betaine aldehyde → betaine). Protoplast-derived chloroplasts of spinach (Spinacia oleracea L.) carry out both reactions, more rapidly in light than in darkness (AD Hanson et al. 1985 Proc Natl Acad Sci USA 82: 3678-3682). We investigated the light-stimulated oxidation of choline, using spinach chloroplasts isolated directly from leaves. The rates of choline oxidation obtained (dark and light rates: 10-50 and 100-300 nanomoles per hour per milligram chlorophyll, respectively) were approximately 20-fold higher than for protoplast-derived chloroplasts. Betaine aldehyde was the main product. Choline oxidation in darkness and light was suppressed by hypoxia. Neither uncouplers nor the Calvin cycle inhibitor glyceraldehyde greatly affected choline oxidation in the light, and maximal choline oxidation was attained far below light saturation of CO₂ fixation. The light stimulation of choline oxidation was abolished by the PSII inhibitors DCMU and dibromothymoquinone, and was partially restored by adding reduced diaminodurene, an electron donor to PSI. Both methyl viologen and phenazine methosulfate prevented choline oxidation. Adding dihydroxyacetone phosphate, which can generate NADPH in organello, doubled the dark rate of choline oxidation. These results indicate that choline oxidation in chloroplasts requires oxygen, and reducing power generated from PSI. Enzymic reactions consistent with these requirements are discussed.     

The Nature of Light-Induced Inhibition of Photosystem II in Pumpkin (Cucurbita pepo L.) Leaves Depends on Temperature

Attached leaves of pumpkin (Cucurbita pepo L.) were treated in high or moderate light at room temperature or a 1°C. The symptoms of photoinhibition appearing during light treatments at room temperature could be attributed to a decrease in the primary activity of PSII. However, when the light treatment was given at 1°C, the quantum yield of photosynthetic oxygen evolution decreased much more than would be expected from the decrease in the ratio of variable to maximum fluorescence at 77°K. Also, light treatment at 1°C lowered the chloroplast wholechain electron transfer capacity much more than it affected PSII electron transport (H₂O to paraphenylbenzoquinone). Light treatments at both room temperature and 1°C led to an increase in B_max, which indicates an increase in the proportion of PSII_β centers. PSI was not affected by the light treatments, and the treatments in the dark at 1°C caused only minor changes in the measured properties of the leaves. We conclude that high light always inhibits the primary activity of PSII, but at low temperature there is greater inhibition of electron transfer from primary electron accepting plastoquinone of PSII to the plastoquinone pool, which leads to a drastic decrease in the quantum yield of oxygen evolution in the chilling-sensitive pumpkin.     

Characterization of the wave phenomenon of flash-induced chlorophyll fluorescence in Chlamydomonas reinhardtii

Flash-induced chlorophyll fluorescence relaxation is a powerful tool to monitor the reoxidation reactions of the reduced primary quinone acceptor, Q_A^? by Q_B and the plastoquinone (PQ) pool, as well as the charge recombination reactions between the donor and acceptor side components of Photosystem II (PSII). Under certain conditions, when the PQ pool is highly reduced (e.g. in microaerobic conditions), a wave phenomenon appears in the fluorescence relaxation kinetics, which reflects the transient reoxidation and re-reduction of Q_A^? by various electron transfer processes, which in cyanobacteria is mediated by NAD(P)H dehydrogenase (NDH-1). The wave phenomenon was also observed and assigned to the operation of type 2 NAD(P)H dehydrogenase (NDH-2) in the green alga Chlamydomonas reinhardtii under hydrogen-producing conditions, which required a long incubation of algae under sulphur deprivation (Krishna et al. J Exp Bot 70 (21):6321–6336, 2019). However, the conditions that induce the wave remained largely uncharacterized so far in microalgae. In this work, we investigated the wave phenomenon in Chlamydomonas reinhardtii under conditions that lead to a decrease of PSII activity by applying hydroxylamine treatment, which impacts the donor side of PSII in combination with a strongly reducing environment of the PQ pool (microaerobic conditions). A similar wave phenomenon could be induced by photoinhibitory conditions (illumination with strong light in the presence of the protein synthesis inhibitor lincomycin). These results indicate that the fluorescence wave phenomenon is activated in green algae when the PSII activity decreases relative to Photosystem I (PS I) activity and the PQ pool is strongly reduced. Therefore, the fluorescence wave could be used as a sensitive indicator of altered intersystem electron transfer processes, e.g. under stress conditions.