Xanthophyll Conversion Research Articles

Lipid Dependence of Xanthophyll Cycling in Higher Plants and Algae.

The xanthophyll cycles of higher plants and algae represent an important photoprotection mechanism. Two main xanthophyll cycles are known, the violaxanthin cycle of higher plants, green and brown algae and the diadinoxanthin cycle of Bacillariophyceae, Xanthophyceae, Haptophyceae, and Dinophyceae. The forward reaction of the xanthophyll cycles consists of the enzymatic de-epoxidation of violaxanthin to antheraxanthin and zeaxanthin or diadinoxanthin to diatoxanthin during periods of high light illumination. It is catalyzed by the enzymes violaxanthin or diadinoxanthin de-epoxidase. During low light or darkness the back reaction of the cycle, which is catalyzed by the enzymes zeaxanthin or diatoxanthin epoxidase, restores the epoxidized xanthophylls by a re-introduction of the epoxy groups. The de-epoxidation reaction takes place in the lipid phase of the thylakoid membrane and thus, depends on the nature, three dimensional structure and function of the thylakoid lipids. As the xanthophyll cycle pigments are usually associated with the photosynthetic light-harvesting proteins, structural re-arrangements of the proteins and changes in the protein-lipid interactions play an additional role for the operation of the xanthophyll cycles. In the present review we give an introduction to the lipid and fatty acid composition of thylakoid membranes of higher plants and algae. We introduce the readers to the reaction sequences, enzymes and function of the different xanthophyll cycles. The main focus of the review lies on the lipid dependence of xanthophyll cycling. We summarize the current knowledge about the role of lipids in the solubilization of xanthophyll cycle pigments. We address the importance of the three-dimensional lipid structures for the enzymatic xanthophyll conversion, with a special focus on non-bilayer lipid phases which are formed by the main thylakoid membrane lipid monogalactosyldiacylglycerol. We additionally describe how lipids and light-harvesting complexes interact in the thylakoid membrane and how these interactions can affect the structure of the thylakoids. In a dedicated chapter we offer a short overview of current membrane models, including the concept of membrane domains. We then use these concepts to present a model of the operative xanthophyll cycle as a transient thylakoid membrane domain which is formed during high light illumination of plants or algal cells.

In vivo photoprotection mechanisms observed from leaf spectral absorbance changes showing VIS\u2013NIR slow-induced conformational pigment bed changes

Regulated heat dissipation under excessive light comprises a complexity of mechanisms, whereby the supramolecular light-harvesting pigment–protein complex (LHC) shifts state from light harvesting towards heat dissipation, quenching the excess of photo-induced excitation energy in a non-photochemical way. Based on whole-leaf spectroscopy measuring upward and downward spectral radiance fluxes, we studied spectrally contiguous (hyperspectral) transient time series of absorbance A(λ,t) and passively induced chlorophyll fluorescence F(λ,t) dynamics of intact leaves in the visible and near-infrared wavelengths (VIS–NIR, 400–800 nm) after sudden strong natural-like illumination exposure. Besides light avoidance mechanism, we observed on absorbance signatures, calculated from simultaneous reflectance R(λ,t) and transmittance T(λ,t) measurements as A(λ,t) = 1 − R(λ,t) − T(λ,t), major dynamic events with specific onsets and kinetical behaviour. A consistent well-known fast carotenoid absorbance feature (500–570 nm) appears within the first seconds to minutes, seen from both the reflected (backscattered) and transmitted (forward scattered) radiance differences. Simultaneous fast Chl features are observed, either as an increased or decreased scattering behaviour during quick light adjustment consistent with re-organizations of the membrane. The carotenoid absorbance feature shows up simultaneously with a major F decrease and corresponds to the xanthophyll conversion, as quick response to the proton gradient build-up. After xanthophyll conversion (t = 3 min), a kinetically slower but major and smooth absorbance increase was occasionally observed from the transmitted radiance measurements as wide peaks in the green (~ 550 nm) and the near-infrared (~ 750 nm) wavelengths, involving no further F quenching. Surprisingly, in relation to the response to high light, this broad and consistent VIS–NIR feature indicates a slowly induced absorbance increase with a sigmoid kinetical behaviour. In analogy to sub-leaf-level observations, we suggest that this mechanism can be explained by a structure-induced low-energy-shifted energy redistribution involving both Car and Chl. These findings might pave the way towards a further non-invasive spectral investigation of antenna conformations and their relations with energy quenching at the intact leaf level, which is, in combination with F measurements, of a high importance for assessing plant photosynthesis in vivo and in addition from remote observations.

The Dynamics of Energy Dissipation and Xanthophyll Conversion in Arabidopsis Indicate an Indirect Photoprotective Role of Zeaxanthin in Slowly Inducible and Relaxing Components of Non-photochemical Quenching of Excitation Energy.

The dynamics of non-photochemical quenching (NPQ) of chlorophyll fluorescence and the dynamics of xanthophyll conversion under different actinic light conditions were studied in intact leaves of Arabidopsis thaliana. NPQ induction was investigated during up to 180 min illumination at 450, 900, and 1,800 μmol photons m−2 s−1 (μE) and NPQ relaxation after 5, 30, 90, or 180 min of pre-illumination at the same light intensities. The comparison of wild-type plants with mutants affected either in xanthophyll conversion (npq1 and npq2) or PsbS expression (npq4 and L17) or lumen acidification (pgr1) indicated that NPQ states with similar, but not identical characteristics are induced at longer time range (15–60 min) in wild-type and mutant plants. In genotypes with an active xanthophyll conversion, the dynamics of two slowly (10–60 min) inducible and relaxing NPQ components were found to be kinetically correlated with zeaxanthin formation and epoxidation, respectively. However, the extent of NPQ was independent of the amount of zeaxanthin, since higher NPQ values were inducible with increasing actinic light intensities without pronounced changes in the zeaxanthin amount. These data support an indirect role of zeaxanthin in pH-independent NPQ states rather than a specific direct function of zeaxanthin as quencher in long-lasting NPQ processes. Such an indirect function might be related to an allosteric regulation of NPQ processes by zeaxanthin (e.g., through interaction of zeaxanthin at the surface of proteins) or a general photoprotective function of zeaxanthin in the lipid phase of the membrane (e.g., by modulation of the membrane fluidity or by acting as antioxidant). The found concomitant down-regulation of zeaxanthin epoxidation and recovery of photosystem II activity ensures that zeaxanthin is retained in the thylakoid membrane as long as photosystem II activity is inhibited or down-regulated. This regulation supports the view that zeaxanthin can be considered as a kind of light stress memory in chloroplasts, allowing a rapid reactivation of photoprotective NPQ processes in case of recurrent light stress periods.

Field assessment of sub-epidermal leaf anthocyanin, PSII photochemistry, and the xanthophyll-cycle as photoprotective mechanisms in two morphs of Agave striata

We tested the hypothesis that leaf epidermal pigments screen light of particular wavelengths from reaching the photosynthetic machinery, reducing dependence on the xanthophyll-cycle as an energy dissipation process. Under field conditions, photosynthesis and water relations were studied in two morphs of Agave striata that differ in leaf coloration (green vs. reddish-purple). Titratable acidity, chlorophyll fluorescence, and internal and surface leaf temperatures were measured under low irradiance by shading (30%) and full sunlight (100%) for six days. We also measured the reflectance ratio (RRED: RGREEN), an index of anthocyanin content and the change in photochemical reflectance index (ΔPRI), an index of xanthophyll-cycle de-epoxidation state (xanthophyll conversion). Our results showed that both morphs expressed typical CAM-activity with no significant differences under sun vs. shade. However, shading did reduce titratable acids in both morphs. Both morphs were well hydrated, with the relative water content (RWC) being greater than 93%. Leaf surface temperature was found to be significantly higher during the day in the green morph compared to the red morph under sun and shade. Dark level fluorescence (Fo), photochemical efficiency of PSII (Fv/Fm), and the quantum yield of PSII electron transport (ΦPSII) were higher in the red morph under sun compared to the green morph. The value of qN (non-photochemical quenching) was significantly higher during the day for the green morph compared to the red morph and this higher qN value was associated with a greater xanthophyll conversion and surface leaf temperature. However, sunlight did not predispose either of the morphs to photoinhibition. It is clear that the sub-epidermal anthocyanins serve as a photoprotective mechanism in the red morph, screening light energy from reaching the photosynthetic machinery and reducing dependence on the xanthophyll-cycle. We concluded that under natural light conditions the leaves of two morphs tested utilized differential photoprotective mechanisms.

The investigation of violaxanthin de-epoxidation in the primitive green alga Mantoniella squamata (Prasinophyceae) indicates mechanistic differences in xanthophyll conversion to higher plants

Schaller S., Latowski D., JemioŁa-Rzemińska M., Quaas T., Wilhelm C., StrzaŁka K. and Goss R. 2012. The investigation of violaxanthin de-epoxidation in the primitive green alga Mantoniella squamata (Prasinophyceae) indicates mechanistic differences in xanthophyll conversion to higher plants. Phycologia 51: 359–370. DOI: 10.2116/11-127.1The violaxanthin (Vx) de-epoxidation of the green alga Mantoniella squamata (Prasinophyceae) was investigated in thylakoid membranes and in enzyme assays with the isolated Vx de-epoxidase (VDE), where pure Vx or purified light-harvesting complexes of M. squamata (MLHC) or spinach (LHCII) were used. The detailed analysis of the VDE amino acid sequences of prasinophycean algae and higher plants suggests that structural differences of the M. squamata VDE are responsible for the slow conversion of Ax to Zx that is typical for this alga. The present experiments further demonstrate that the availability of the substrate Vx has a strong impact on the overall de-epoxidation reaction and the ratio of the second de-epoxidation rate (de-epoxidation from Ax to Zx) to the first de-epoxidation rate (conversion from Vx to Ax). High concentrations of available Vx, as found in enzyme assays with pure Vx, lead to saturation of the VDE and a strong competition with the intermediate reaction product Ax, thus decreasing the ratio of the second de-epoxidation rate to the first de-epoxidation rate. In de-epoxidation assays with isolated LHCs, where the availability of Vx is limited by the detachment of Vx from the protein, an increased ratio of the second de-epoxidation rate to the first de-epoxidation rate is observed. The present study also shows that the interaction between the isolated LHC and the VDE can influence the ratio of the two de-epoxidation rates. Mantoniella squamata VDE, which exhibits a very low ratio of the second de-epoxidation rate to the first de-epoxidation rate in thylakoids or in enzyme assays with the MLHC, shows a high ratio when it is incubated with the LHCII of spinach.

The effects of moderately high temperature on zeaxanthin accumulation and decay

Moderately high temperature reduces photosynthetic capacities of leaves with large effects on thylakoid reactions of photosynthesis, including xanthophyll conversion in the lipid phase of the thylakoid membrane. In previous studies, we have found that leaf temperature of 40°C increased zeaxanthin accumulation in dark-adapted, intact tobacco leaves following a brief illumination, but did not change the amount of zeaxanthin in light-adatped leaves. To investigate heat effects on zeaxanthin accumulation and decay, zeaxanthin level was monitored optically in dark-adapted, intact tobacco and Arabidopsis thaliana leaves at either 23 or 40°C under 45-min illumination. Heated leaves had more zeaxanthin following 3-min light but had less or comparable amounts of zeaxanthin by the end of 45min of illumination. Zeaxanthin accumulated faster at light initiation and decayed faster upon darkening in leaves at 40°C than leaves at 23°C, indicating that heat increased the activities of both violaxanthin de-epoxidase (VDE) and zeaxanthin epoxidase (ZE). In addition, our optical measurement demonstrated in vivo that weak light enhances zeaxanthin decay relative to darkness in intact leaves of tobacco and Arabidopsis, confirming previous observations in isolated spinach chloroplasts. However, the maximum rate of decay is similar for weak light and darkness, and we used the maximum rate of decay following darkness as a measure of the rate of ZE during steady-state light. A simulation indicated that high temperature should cause a large shift in the pH dependence of the amount of zeaxanthin in leaves because of differential effects on VDE and ZE. This allows for the reduction in ΔpH caused by heat to be offset by increased VDE activity relative to ZE.

Diel variation of chlorophyll- a fluorescence, phytoplankton pigments and productivity in the Sub-Antarctic and Polar Front Zones south of Tasmania, Australia

Marine primary production is a fundamental measure of the ocean's capacity to convert carbon dioxide to particulate organic carbon for the marine foodweb, and as such is an essential variable used in ecosystem and biogeochemical models to assess trophic dynamics and carbon cycling. The Sub-Antarctic Zone (SAZ) is a major sink for atmospheric carbon and exhibits large gradients in ocean conditions on both temporal and spatial scales. In this dynamic system, an understanding of small-scale temporal changes is critical for modelling primary production at larger scales. Thus, we investigated diel effects on maximum quantum yield of PSII ( F V/ F M), photosynthetic pigment pools and primary productivity in the western (Diel 1) and eastern SAZ region (Diel 3) south of Tasmania, Australia, and compared this to a station at the polar front (Diel 2). Phytoplankton in the eastern SAZ had the greatest diel response, with cells showing decreased F V/ F M and increased biosynthesis and transformation of xanthophyll and other photoprotective pigments during the day, but only in the surface waters (0 and 10 m). Diel responses diminished by 30 m. Cells in the western SAZ had similar responses across the depths sampled, increasing their F V/ F M during the night and increasing their xanthophyll pigment content during the day. Phytoplankton at the polar front (Diel 2) showed intermediate diel-related variations in photophysiology, with xanthophyll conversion and increases in photoprotective pigments during the day but constant F V/ F M. These diel changes at all sampling stations had little impact on carbon fixation rates, although cells sampled from the deep chlorophyll maximum at the polar front had significantly lower maximum carbon fixation and minimum saturating irradiance ( E k ) compared to the other depths and stations. Considering the oceanographic context, cells at Diel 1 and 2 received less light and were more deeply mixed than cells at Diel 3, causing a dampening of the diel response. These results highlight that phytoplankton in the SAZ is regulated by the physical processes of mixing and light provision, but short-term diel effects on maximum quantum yield of PSII and photoprotective pigments may not propagate to changes in carbon fixation, particularly when cells are nutrient replete. If however, the more stratified eastern SAZ (which had the greatest diel responses) is indicative of how the SAZ region might respond to climate change, then diel effects may become more prominent in the future.

Photoprotective mechanisms during leaf ontogeny: cuticular development and anthocyanin deposition in two morphs of Agave striata that differ in leaf coloration

Our objectives were to test whether in Agave striata Zucc., a plant with crassulacean acid metabolism (CAM plant), leaf wax development is a delayed response to sunlight exposure following cutin development, and whether energy dissipation shifts from non-photochemical quenching to photochemical quenching during leaf ontogeny. Under field conditions, photosynthesis, cuticular development, and anthocyanin deposition were studied in two morphs of A. striata that differ in leaf coloration (green vs. red). We quantified leaf anthocyanin, wax, and cutin content, and also measured chlorophyll a fluorescence and leaf surface temperature. In addition, using three leaf reflectance indices, we measured relative chlorophyll and anthocyanin content, and also xanthophyll-cycle de-epoxidation state (xanthophyll conversion). Our results revealed that the main components of cuticle (wax and cutin) in leaves of A. striata are deposited during different developmental windows, which are similar to leaves of monocots such as grasses. Exposure to sunlight was found to be the most likely candidate to affect wax and anthocyanin deposition. Chlorophyll a fluorescence data revealed that the sunlight conditions experienced by both morphs predisposed the young leaves of the green morph and old leaves of both morphs to photoinhibition. Our results also revealed that old leaves of the red morph, which contain a reduced level of chlorophyll and anthocyanin, had additional photoprotection via xanthophyll conversion. The results presented here support the photoprotective function of leaf anthocyanins and wax accumulation during leaf ontogeny, indicating that their presence may compensate for the reduced dependence of non-photochemical quenching and the xanthophyll-cycle pigment conversion.

De-epoxidation of Violaxanthin in the Minor Antenna Proteins of Photosystem II, LHCB4, LHCB5, and LHCB6

The conversion of violaxanthin to zeaxanthin is essentially required for the pH-regulated dissipation of excess light energy in the antenna of photosystem II. Violaxanthin is bound to each of the antenna proteins of both photosystems. Former studies with recombinant Lhcb1 and different Lhca proteins implied that each antenna protein contributes specifically to violaxanthin conversion related to protein-specific affinities of the different violaxanthin binding sites. We investigated the violaxanthin de-epoxidation in the minor antenna proteins of photosystem II, Lhcb4-6. Recombinant proteins were reconstituted with different xanthophyll mixtures to study the conversion of violaxanthin at different xanthophyll binding sites in these proteins. The extent and kinetics of violaxanthin de-epoxidation were found to be dependent on the respective protein and, for each protein, also on the binding site of violaxanthin. In particular, violaxanthin bound to Lhcb4 was nearly inconvertible for de-epoxidation, whereas violaxanthin bound to Lhcb5 was fully convertible but with slow kinetics. Lhcb6 exhibited heterogeneous violaxanthin conversion characteristics, which could be assigned to different populations of reconstituted Lhcb6 complexes with respect to violaxanthin binding sites. The results support the proposed different binding affinities of violaxanthin to the three putative violaxanthin binding sites (V1, N1, and L2) in antenna proteins. Under consideration of former studies with Lhcb1 and Lhca proteins, the data imply that violaxanthin bound to the V1 and N1 binding site of antenna proteins is easily accessible for de-epoxidation in all antenna proteins, whereas violaxanthin bound to L2 is either only slowly or not convertible to zeaxanthin, depending on the respective protein.

Leaf angle responds to nitrogen supply in eucalypt seedlings. Is it a photoprotective mechanism?

We examined the adjustment of leaf angle (L theta) and foliar chlorophyll and xanthophyll chemistry in Eucalyptus nitens (Deane and Maiden) Maiden seedlings maintained in various nitrogen (N)-supply treatments over a 6-month period. Adjustment of L theta toward the vertical was greatest under conditions of foliar N deficiency and became incrementally more horizontal with increasing foliar N concentration. Photochemical efficiency (Fv/Fm) and quantum yield were lower in seedlings with low foliar N (low-N seedlings) in winter, but not in autumn. Low-N seedlings generally had low area-based chlorophyll concentrations and high xanthophyll-cycle conversion ratios, particularly during months of low temperature. Under mild temperature conditions, high concentrations of zeaxanthin and antheraxanthin were associated with lower electron transport rates (ETR). Incident light, Fv/Fm, ETR and total chlorophyll concentration were negatively correlated with L theta, with horizontal leaf orientation measured as 0 degrees and vertical leaf orientation as 90 degrees . Xanthophyll conversion ratio was positively correlated with L theta. Adjustments in L theta may play a role in photoprotection of E. nitens seedlings by assisting the leaf to balance its utilization and dissipation of energy.

Photoacclimation modulates excessive photosynthetically active and ultraviolet radiation effects in a temperate and an Antarctic marine diatom

The influence of photoacclimation on the effects of excessive photosynthetically active (PAR; 400-700 nm) and ultraviolet (UVR; 280-400 nm) radiation was assessed for the marine diatoms Thalassiosira weissflogii (Grunow) Fryxell and Hasle and Thalassiosira antarctica (Comber). Low and high PAR acclimated cultures were subjected to simulated surface irradiance (SSI) that mimicked irradiance around noon, including UVR. PSII efficiency, xanthophyll conversion, superoxide dismutase (SOD) activity, carbohydrate buildup, and lipid peroxidation were investigated after 30 min SSI and during 120 min recovery in low irradiance. Furthermore, viability loss was measured during 4 h SSI. Prior to SSI, the diadino-diatoxanthin pool was increased in high irradiance acclimated cells, compared with cells grown under low irradiance. Thirty-minutes SSI caused a pronounced decline in PSII efficiency. This coincided with de-epoxidation of diadinoxanthin in high irradiance acclimated cells, which was completely reversed during recovery in low irradiance. De-epoxidation was lower for low irradiance acclimated cells, whereas PSII efficiency and carbohydrate buildup were lower during the recovery phase. Furthermore, clear UVR effects on PSII efficiency were observed in low irradiance but not in high irradiance acclimated cells. Although 30 min SSI did not increase cellular SOD activity and lipid peroxidation, prolonged (4 h) SSI caused viability loss in low irradiance acclimated cells, which was enhanced by UVR. Therefore, PAR and UVR-induced PSII inactivation and viability loss were reduced by high irradiance-mediated changes in light harvesting and the xanthophyll pigments. In addition to photoacclimation-modulated differences, minor sensitivity differences were found between species.

De-epoxidation of Violaxanthin in Light-harvesting Complex I Proteins

The conversion of violaxanthin (Vx) to zeaxanthin (Zx) in the de-epoxidation reaction of the xanthophyll cycle plays an important role in the protection of chloroplasts against photooxidative damage. Vx is bound to the antenna proteins of both photosystems. In photosystem II, the formation of Zx is essential for the pH-dependent dissipation of excess light energy as heat. The function of Zx in photosystem I is still unclear. In this work we investigated the de-epoxidation characteristics of light-harvesting complex proteins of photosystem I (LHCI) under in vivo and in vitro conditions. Recombinant LHCI (Lhcal-4) proteins were reconstituted with Vx and lutein, and the convertibility of Vx was studied in an in vitro assay using partially purified Vx de-epoxidase isolated from spinach thylakoids. All four LHCI proteins exhibited unique de-epoxidation characteristics. An almost complete Vx conversion to Zx was observed only in Lhca3, whereas Zx formation in the other LHCI proteins decreased in the order Lhca4 > Lhca1 > Lhca2. Most likely, these differences in Vx de-epoxidation were related to the different accessibility of the respective carotenoid binding sites in the distinct antenna proteins. The results indicate that Vx bound to site V1 and N1 is easily accessible for de-epoxidation, whereas Vx bound to L2 is only partially and/or with the slower kinetics convertible to Zx. The de-epoxidation properties determined for the monomeric recombinant proteins were consistent with those obtained for isolated native LHCI-730 and LHCI-680 in the same in vitro assay and the de-epoxidation state found under in vivo conditions in native LHCIs.

The kinetics of zeaxanthin formation is retarded by dicyclohexylcarbodiimide

The de-epoxidation of violaxanthin to antheraxanthin (Anth) and zeaxanthin (Zeax) in the xanthophyll cycle of higher plants and the generation of nonphotochemical fluorescence quenching in the antenna of photosystem II (PSII) are induced by acidification of the thylakoid lumen. Dicyclohexylcarbodiimide (DCCD) has been shown (a) to bind to lumen-exposed carboxy groups of antenna proteins and (b) to inhibit the pH-dependent fluorescence quenching. The possible influence of DCCD on the de-epoxidation reactions has been investigated in isolated pea (Pisum sativum L.) thylakoids. The Zeax formation was found to be slowed down in the presence of DCCD. The second step (Anth --> Zeax) of the reaction sequence seemed to be more affected than the violaxanthin --> Anth conversion. Comparative studies with antenna-depleted thylakoids from plants grown under intermittent light and with unstacked thylakoids were in agreement with the assumption that binding of DCCD to antenna proteins is probably responsible for the retarded kinetics. Analyses of the DCCD-induced alterations in different antenna subcomplexes showed that Zeax formation in the PSII antenna proteins was predominantly influenced by DCCD, whereas Zeax formation in photosystem I was nearly unaffected. Our data support the suggestion that DCCD binding to PSII antenna proteins is responsible for the observed alterations in xanthophyll conversion.

Comparison of chlorophyll fluorescence quenching in leaves of wild-type with a chlorophyll-b-less mutant of barley ( Hordeum vulgare L.)

A pulse amplitude-modulated fluorometric technique was employed to separate photochemical (qP) and non-photochemical (qN) chlorophyll fluorescence quenching in attached leaves of wild-type and a chlorophyll-b-less mutant of barley ( Hordeum vulgare L.). Whereas a significantly higher qN developed in the wild type in the intensity range at which the photosynthetic light response became non-linear, there were virtually no differences in qP until pronounced photoinhibition was apparent. On monitoring the “dark” recovery of the maximum ( F M) and dark level ( F 0) fluorescence yield, three distinct kinetic phases were resolved, which are ascribed to relaxation of high energy state quenching (qE), state 1-state 2 transitions (qT) and quenching due to photoinhibition (qI). The results provide evidence for heterogeneity of qE. The major part of qE (related to a fast-relaxing phase of F M quenching) is strongly reduced in mutant leaves. A fast-relaxing phase of F 0 quenching, readily observed in wild-type leaves under high light, is absent in the mutant under the same conditions. Hence, qE appears to be associated with the photosystem II light-harvesting complex (LHC II). A medium component of “dark” relaxation kinetics was observed in both mutant and wild-type leaves. However, it appears attributable to qT only in the latter at low light. Supported by the finding that there was no difference in xanthophyll pool size (on a chlorophyll a basis) between wild type and mutant, under high light conditions the medium phase may reflect a slower-relaxing portion of qE, probably due to xanthophyll conversion. Under supersaturating light the very slowly relaxing qI component became dominating, affecting mutant leaves to a considerably greater extent. The results stress the key role of LHC II not only as a solar energy collector but also in protecting the photosynthetic apparatus from adverse effects of excess excitation input.

Carotenoid transformations underlying the blue absorbance change in flashed leaves during the induction of oxygen evolution

The blue absorbance change occurring in flashed bean (Phaseolus vulgaris L.) leaves when exposed to continuous light (first observed by Strasser; Strasser, R.J. (1973) Arch. Int. Physiol. Biochem. 81, 935–955) is caused by the conversion of the following xanthophylls: violaxanthine → antheraxanthine → zeaxanthine. This conclusion is derived from the simultaneous occurrence of both reactions: (a) In flashed leaves, blue absorbance change and xanthophyll conversion take place under strong (2 mW · cm−2) but not under weak (0.02 mW · cm−2) white light. (b) In chloroplasts isolated from flashed leaves, the blue absorbance change occurs in the dark under conditions that also induce the xanthophyll conversion. (c) Blue absorbance change and xanthophyll conversion are both inhibited by dithiothreitol. In addition, the light-induced blue absorbance change is reversed in the dark if aerobic conditions are maintained, i.e. under conditions that in normal leaves favor the reversal of the above reaction sequence.The significance of the xanthophyll conversion is discussed in relation to other phenomena occurring in flashed leaves after exposure to continuous illumination.