Nocturnally retained zeaxanthin does not remain engaged in a state primed for energy dissipation during the summer in two Yucca species growing in the Mojave Desert

Differently oriented leaves of Yucca schidigera and Yucca brevifolia were characterized in the Mojave Desert with respect to photosystem II and xanthophyll cycle activity during three different seasons, including the hot and dry summer, the relatively cold winter, and the mild spring season. Photosynthetic utilization of a high percentage of the light absorbed in PSII was observed in all leaves only during the spring, whereas very high levels of photoprotective, thermal energy dissipation were employed both in the summer and the winter season in all exposed leaves of both species. Both during the summer and the winter season, when energy dissipation levels were high diurnally, xanthophyll cycle pools (relative to either Chl or other carotenoids) were higher relative to the spring, and a nocturnal retention of high levels of zeaxanthin and antheraxanthin (Z + A) occurred in all exposed leaves of both species. Although this nocturnal retention of Z + A was associated with nocturnal maintenance of a low PSII efficiency (F_v/F_m) on a cold winter night, pre‐dawn F_v/F_m was high in (Z + A)‐retaining leaves following a warm summer night. This indicates nocturnal engagement of Z + A in a state primed for energy dissipation throughout the cold winter night – while high levels of retained Z + A were not engaged for energy dissipation prior to sunrise on a warm summer morning. Possible mechanisms for a lack of sustained engagement of retained Z + A for energy dissipation at elevated temperatures are discussed.     

Xanthophyll cycle components and capacity for non-radiative energy dissipation in sun and shade leaves ofLigustrum ovalifolium exposed to conditions limiting photosynthesis

The relationships between photosynthetic efficiency, non-radiative energy dissipation and carotenoid composition were studied in leaves ofLigustrum ovalifolium developed either under full sunlight or in the shade. Sun leaves contained a much greater pool of xanthophyll cycle components than shade leaves. The rate of non-radiative energy dissipation, measured as non-photochemical fluorescence quenching (NPQ), was strictly related to the deepoxidation state (DPS) of xanthophyll cycle components in both sun and shade leaves, indicating that zeaxanthin (Z) and antheraxanthin (A) are involved in the development of NPQ. Under extreme conditions of excessive energy, sun leaves showed higher maximum DPS than shade leaves. Therefore, sun leaves contained not only a greater pool of xanthophyll cycle components but also a higher proportion of violaxanthin (V) actually photoconvertible to A and Z, compared to shade leaves. Both these effects contributed to the higher NPQ in sun versus shade leaves. The amount of photoconvertible V was strongly related to chla/b ratio and inversely to leaf neoxanthin content. This evidence indicates that the amount of photoconvertible V may be dependent on the degree of thylakoid membrane appression and on the organization of chlorophyll-protein complexes, and possible explanations are discussed. Exposure to chilling temperatures caused a strong decline in the photon yield of photosynthesis and in the intrinsic efficiency of PS II photochemistry in sun leaves, but little effects in shade leaves. These effects were accompanied by increases in the pool of xanthophyll cycle components and in DPS, more pronounced in sun than in shade leaves. This corroborates the view that Z and A may play a photoprotective role under unfavorable conditions. In addition to the xanthophyll-related non-radiative energy dissipation, a slow relaxing component of NPQ, independent from A and Z concentrations, has been found in leaves exposed to low temperature and high light. This quenching component may be attributed either to other regulatory mechanism of PS II efficiency or to photoinactivation.Research supported by National Research Council of Italy, Special Project RAISA, Sub-Project 2, Paper N. 1587.     

The xanthophyll cycle and energy dissipation in differently oriented faces of the cactus Opuntia macrorhiza

  Diurnal changes in titratable acidity, photosynthesis, energy dissipation activity, and the carotenoid composition of differently oriented cladodes of the cactus Opuntia macrorhiza were characterized during exposure to full sunlight in the field. Four cladode faces were chosen such that each was exposed to maximum photon flux densities (PFD) at different times of the day in addition to receiving different daily integrated PFDs. The sum of all carotenoids per chlorophyll was found to increase with increasing exposure to PFD, with the carotenoids of the xanthophyll cycle present in the most exposed face at more than twice the concentration found in the least exposed face. All faces exhibited large increases in xanthophyll cycle-dependent energy dissipation as the sun rose in the morning, even those receiving only minimal levels of diffuse radiation. The transient high levels of energy dissipation in those faces that did not receive direct sunlight in the morning may have been due to low temperature inhibition of photosynthesis (predawn low of 2°C). For the two faces receiving peak PFDs in the morning hours (north and east faces), the level of energy dissipation activity increased rapidly during exposure to direct sunlight in the early morning, gradually declining in the late morning under warm temperatures, and was negligible during the afternoon low light conditions. Changes in the xanthophyll cycle paralleled the changes in energy dissipation with the majority of the cycle present as violaxanthin (V) prior to sunrise, largely de-epoxidized to zeaxanthin (Z) and antheraxanthin (A) during exposure to direct sunlight, and reconverted to V during the afternoon. For the two faces receiving peak PFDs in the afternoon (south and west faces), energy dissipation activity increased dramatically during the early morning low light period, subsequently decreasing during midday as decarboxylation of malic acid proceeded maximally (providing a high concentration of CO₂ for photosynthesis), and then increased to the highest level in the late afternoon as the supply of malic acid was depleted and rates of photosynthetic electron transport declined. The xanthophyll cycle, largely present as Z and A prior to sunrise in the south and west faces, was de-epoxidized to the greatest extent in the late afternoon, followed by epoxidation back to the predawn level by sunset. In all cladode faces high levels of energy dissipation activity were accompanied by decreases in the intrinsic efficiency of photosystem II (PSII), indicative of a regulatory process that diverted the excess energy away from the reaction centers during periods of excess light. Furthermore, the overnight retention of Z and A by the south and west faces was accompanied by a sustained reduction in PSII efficiency (i.e., “photoinhibition”). We suggest that this “photoinhibition” represents the sustained engagement of nocturnally retained Z and A in the photoprotective down-regulation of PSII. Received: 8 May 1996 / Accepted: 9 September 1996     

Internal and external photoprotection in developing leaves of the CAM plant Cotyledon orbiculata

Leaves of the CAM plant Cotyledon orbiculata produced a dense epidermal wax which decreased the absorption of light, possibly functioning as an external photoprotective mechanism (Robinson et al. 1993). However, developing leaves did not accumulate wax until after 21 d with full wax coating not achieved until at least 35 d. In addition, young leaves had lower rates of electron transport than mature leaves. Leaf development therefore occurs at higher incident PFD than that experienced by the mature leaves, and, for young leaves, can lead to an increase in the proportion of light energy which is excess to requirements and must be dissipated non-photochemically. Changes in the photosynthetic capacity, PSII efficiency, rate of energy dissipation, and the content of chlorophyll (Chi), carotenoids, wax and anthocyanins were followed in developing leaves of C. orbiculata in an attempt to elucidate the relative importance of the various photoprotective mechanisms during leaf ontogeny. The largest pools of xanthophyll cycle pigments (on a Chi basis) were found in the waxless, young leaves and were correlated with greater levels of energy dissipation activity. The importance of xanthophyll cycle-dependent energy dissipation in young C. orbiculata leaves prior to development of a reflective wax covering, and full photosynthetic capacity which for CAM plants includes appreciable nocturnal acid accumulation, is discussed. Also, we consider the possibility that anthocyanin pigments in the upper and lower epidermis may increase reflectivity and act as external photoprotectants.     

Dependence of photosynthesis and energy dissipation activity upon growth form and light environment during the winter

Two very distinctive responses of photosynthesis to winter conditions have been identified. Mesophytic species that continue to exhibit growth during the winter typically exhibit higher maximal rates of photosynthesis during the winter or when grown at lower temperatures compared to individuals examined during the summer or when grown at warmer temperatures. In contrast, sclerophytic evergreen species growing in sun-exposed sites typically exhibit lower maximal rates of photosynthesis in the winter compared to the summer. On the other hand, shaded individuals of those same sclerophytic evergreen species exhibit similar or higher maximal rates of photosynthesis in the winter compared to the summer. Employment of the xanthophyll cycle in photoprotective energy dissipation exhibits similar characteristics in the two groups of plants (mesophytes and shade leaves of sclerophytic evergreens) that exhibit upregulation of photosynthesis during the winter. In both, zeaxanthin + antheraxanthin (Z + A) are retained and PS II remains primed for energy dissipation only on nights with subfreezing temperatures, and this becomes rapidly reversed upon exposure to increased temperatures. In contrast, Z + A are retained and PS II remains primed for energy dissipation over prolonged periods during the winter in sun leaves of sclerophytic evergreen species, and requires days of warming to become fully reversed. The rapid disengagement of this energy dissipation process in the mesophytes and shade sclerophytes apparently permits a rapid return to efficient photosynthesis and increased activity on warmer days during the winter. This may be associated with a decreasing opportunity for photosynthesis in source leaves relative to the demand for photosynthesis in the plant's sinks. In contrast, the sun-exposed sclerophytes – with a relatively high source to sink ratio – maintain PS II in a state primed for high levels of energy dissipation activity throughout much of the winter. Independent of whether photosynthesis was up- or downregulated, all species under all conditions exhibited higher levels of soluble carbohydrates during the winter compared to the summer. Thus downregulation of photosynthesis and of Photosystem II do not appear to limit carbohydrate accumulation under winter conditions. A possible signal communicating an altered source/sink balance, or that may be influencing the engagement of Z + A in energy dissipation, is phosphorylation of thylakoid proteins such as D1.This revised version was published online in October 2005 with corrections to the Cover Date.     

Seasonal changes in leaf antioxidant systems and xanthophyll cycle characteristics in Taxus x media growing in sun and shade environments

We examined differences between summer and winter in xanthophyll cycle-dependent energy dissipation and leaf antioxidant systems in needles of the overwintering evergreen Taxus x media cv. Tauntonii (Taunton yew) growing in both sun and shade environments in Saint Paul, Minnesota. During the winter, both sun and shade plants exhibited increases in the capacity for, and utilization of, xanthophyll cycle-dependent thermal energy dissipation. Winter needles showed decreases (sun needles) or no change (shade needles) in superoxide dismutase activity (EC 1.15.1.1), no change in ascorbate peroxidase activity (EC 1.11.1.11) and no change (sun needles) or increases (shade needles) in reduced ascorbate levels. Both sun and shade needles showed large increases in glutathione reductase activity (EC 1.6.4.2) and total glutathione levels during the winter, in addition to increases in levels of α-tocopherol. These results suggest an important photoprotective role during the winter for xanthophyll cycle-dependent energy dissipation and for the antioxidants glutathione and α-tocopherol. They suggest a less important photoprotective function of the enzyme-based water–water cycle in winter acclimation in the seasonally very cold environment of Minnesota.     

田间大豆叶片成长过程中的光合特性及光破坏防御机制

田间大豆叶片在成长进程中光饱和光合速率持续提高,但气孔导度的增加明显滞后.尽管叶片在成长初期就具有较高的最大光化学效率,但是仍略低于发育成熟的叶片.随着叶片的成长,光下叶片光系统Ⅱ实际效率增加;非光化学猝灭下降.幼叶叶黄素总量与叶绿素之比较高,随着叶面积的增加该比值下降,在光下,幼叶的脱环氧化程度较高.因此认为大豆叶片成长初期就能够有效地进行光化学调节;在叶片生长过程中依赖叶黄素循环的热耗散机制迅速建立起来有效抵御强光的破坏.     

Mechanistic aspects of xanthophyll cycle-dependent photoprotection in higher plant chloroplasts and leaves

Higher plants must dissipate absorbed light energy that exceeds the photosynthetic capacity to avoid molecular damage to the pigments and proteins that comprise the photosynthetic apparatus. Described in this minireview is a current view of the biochemical, biophysical and bioenergetic aspects of the primary photoprotective mechanism responsible for dissipating excess excitation energy as heat from photosystem II (PSII). The photoprotective heat dissipation is measured as nonphotochemical quenching (NPQ) of the PSII chlorophyll a (Chl a) fluorescence. The NPQ mechanism is controlled by the trans-thylakoid membrane pH gradient (ΔpH) and the special xanthophyll cycle pigments. In the NPQ mechanism, the de-epoxidized endgroup moieties and the trans-thylakoid membrane orientations of antheraxanthin (A) and zeaxanthin (Z) strongly affect their interactions with protonated chlorophyll binding proteins (CPs) of the PSII inner antenna. The CP protonation sites and steps are influenced by proton domains sequestered within the proteo-lipid core of the thylakoid membrane. Xanthophyll cycle enrichment around the CPs may explain why changes in the peripheral PSII antenna size do not necessarily affect either the concentration of the xanthophyll cycle pigments on a per PSII unit basis or the NPQ mechanism. Recent time-resolved PSII Chi a fluorescence studies suggest the NPQ mechanism switches PSII units to an increased rate constant of heat dissipation in a series of steps that include xanthophyll de-epoxidation, CP-protonation and binding of the xanthophylls to the protonated CPs; the concerted process can be described with a simple two-step, pH-activation model. The xanthophyll cycle-dependent NPQ mechanism is profoundly influenced by temperatures suboptimal for photosynthesis via their effects on the trans-thylakoid membrane energy coupling system. Further, low temperature effects can be grouped into either short term (minutes to hours) or long term (days to seasonal) series of changes in the content and composition of the PSII pigment-proteins. This minireview concludes by briefly highlighting primary areas of future research interest regarding the NPQ mechanism.     

Rapid changes in xanthophyll cycle-dependent energy dissipation and photosystem II efficiency in two vines, Stephania japonica and Smilax australis, growing in the understory of an open Eucalyptus forest

Leaves of Stephania japonica and Smilax australis were characterized in situ on the coast of north-eastern New South Wales, Australia, where they were growing naturally in three different light environments: deep shade, in the understory of an open Eucalyptus forest where they received frequent sunflecks of high intensity, and in an exposed site receiving full sunlight. In deep shade the xanthophyll cycle remained epoxidized during the day and the vast majority of absorbed light was utilized for photosynthesis. In the exposed site both deepoxidation and epoxidation of the xanthophyll cycle and changes in the level of xanthophyll-dependent thermal energy dissipation largely tracked the diurnal changes in photon flux density (PFD). In the understory the xanthophyll cycle became largely deepoxidized to zeaxanthin and antheraxanthin upon exposure of the leaves to the first high intensity sunfleck and this high level of deepoxidation was maintained throughout the day both during and between subsequent sunflecks. In contrast, thermal energy dissipation activity, and the efficiency of photosystem II, fluctuated rapidly in response to the changes in incident PFD. These findings suggest a fine level of control over the engagement of zeaxanthin and antheraxanthin in energy dissipation activity, presumably through rapid changes in thylakoid acidification, such that they became rapidly engaged for photoprotection during the sunflecks and rapidly disengaged upon return to low light when continued engagement might limit carbon gain.     

High Contents of Anthocyanins in Young Leaves are Correlated with Low Pools of Xanthophyll Cycle Components and Low Risk of Photoinhibition

We checked the hypothesis that the transient presence of anthocyanins in young leaves serves a photoprotective function. For this purpose, Rosa sp. and Ricinus communis L., whose young leaves are red to become green upon maturation, were used. Thus, young leaves with high and mature leaves with low anthocyanin contents were analysed concerning their carotenoid (Car) composition and susceptibility to photoinhibition. Cars, including the components of the xanthophyll cycle, had similar contents in young and mature leaves, when expressed on a chlorophyll basis. Yet, when expressed on a leaf area basis or on the assumed photon absorptive capacity of leaves, Cars contents were considerably lower in anthocyanic young leaves. Although this may indicate a low photodissipative potential, red young leaves were considerably less susceptible to photoinhibitory damage. The results are compatible with a photoprotective function of anthocyanins, indicating also that their presence may compensate for a low capacity in the xanthophyll cycle-dependent harmless dissipation of excess excitation energy.     

Analysis of xanthophyll cycle carotenoids and chlorophyll fluorescence in light intensity-dependent chlorophyll-deficient mutants of wheat and barley

Three light intensity-dependent Chl b-deficient mutants, two in wheat and one in barley, were analyzed for their xanthophyll cycle carotenoids and Chl fluorescence characteristics under two different growth PFDs (30 versus 600 mol photons·m^–2 s^–1 incident light). Mutants grown under low light possessed lower levels of total Chls and carotenoids per unit leaf area compared to wild type plants, but the relative proportions of the two did not vary markedly between strains. In contrast, mutants grown under high light had much lower levels of Chl, leading to markedly greater carotenoid to Chl ratios in the mutants when compared to wild type. Under low light conditions the carotenoids of the xanthophyll cycle comprised approximately 15% of the total carotenoids in all strains; under high light the xanthophyll cycle pool increased to over 30% of the total carotenoids in wild type plants and to over 50% of the total carotenoids in the three mutant strains. Whereas the xanthophyll cycle remained fairly epoxidized in all plants grown under low light, plants grown under high light exhibited a considerable degree of conversion of the xanthophyll cycle into antheraxanthin and zeaxanthin during the diurnal cycle, with almost complete conversion (over 90%) occurring only in the mutants. 50 to 95% of the xanthophyll cycle was retained as antheraxanthin and zeaxanthin overnight in these mutants which also exhibited sustained depressions in PS II photochemical efficiency (F_v/F_m), which may have resulted from a sustained high level of photoprotective energy dissipation activity. The relatively larger xanthophyll cycle pool in the Chl b-deficient mutant could result in part from the reported concentration of the xanthophyll cycle in the inner antenna complexes, given that the Chl b-deficient mutants are deficient in the peripheral LHC-II complexes.Abbreviations A antheraxanthin - Chl chlorophyll - F_o and F_m minimal yield (at open PS II reaction centers) and maximal yield (at closed centers) of chlorophyll fluorescence in darkness - F level of fluorescence during illumination with photosynthetically active radiation - F_m maximal yield (at closed centers) of chlorophyll fluorescence during illumination with photosynthetically active radiation - (F_m–F)/F_m actual efficiency of PS II during illumination with photosynthetically active radiation - F_v/F_m+(F_m–F_o)/F_m intrinsic efficiency of PS II in darkness - LHC_II light-harvesting chlorophyll-protein complex of Photosystem II - PFD photon flux density (between 400 and 700 nm) - PS I Photosystem I - PS II Photosystem II - V violaxanthin - Z zeaxanthin     

Photoinhibition and photoprotection in senescent leaves of field-grown wheat plants

Photosynthesis, photosystem II (PSII) photochemistry, photoinhibition and the xanthophyll cycle in the senescent flag leaves of wheat (Triticum aestivum L.) plants grown in the field were investigated. Compared to the non-senescent leaves, photosynthetic capacity was significantly reduced in senescent flag leaves. The light intensity at which photosynthesis was saturated also declined significantly. The light response curves of PSII photochemistry indicate that a down-regulation of PSII photochemistry occurred in senescent leaves in particular at high light. The maximal efficiency of PSII photochemistry in senescent flag leaves decreased slightly when measured at predawn but substantially at midday, suggesting that PSII function was largely maintained and photoinhibition occurred in senescent leaves when exposed to high light. At midday, PSII efficiency, photochemical quenching and the efficiency of excitation capture by open PSII centers decreased considerably, while non-photochemical quenching increased significantly. Moreover, compared with the values at early morning, a greater decrease in CO₂ assimilation rate was observed at midday in senescent leaves than in control leaves. The levels of antheraxanthin and zeaxanthin via the de-epoxidation of violaxanthin increased in senescent flag leaves from predawn to midday. An increase in the xanthophyll cycle pigments relative to chlorophyll was observed in senescent flag leaves. The results suggest that the xanthophyll cycle was activated in senescent leaves due to the decrease in CO₂ assimilation capacity and the light intensity for saturation of photosynthesis and that the enhanced formation of antheraxanthin and zeaxanthin at high light may play an important role in the dissipation of excess light energy and help to protect photosynthetic apparatus from photodamage. Our results suggest that the well-known function of the xanthophyll cycle to safely dissipate excess excitation energy is also important for maintaining photosynthetic function during leaf senescence.     

The photoprotective role of epidermal anthocyanins and surface pubescence in young leaves of grapevine (Vitis vinifera)

BACKGROUND AND AIMS: Depending on cultivar, surfaces of young leaves of Vitis vinifera may be glabrous-green ('Soultanina') or transiently have anthocyanins ('Siriki') or pubescence ('Athiri'). A test is made of the hypothesis that anthocyanins and pubescence act as light screens affording a photoprotective advantage to the corresponding leaves, and an assessment is made of the magnitude of their effect. METHODS: Measurements were made on young leaves of the three cultivars in spring under field conditions. Photosynthetic gas-exchange and in vivo chlorophyll fluorescence were measured. Photosynthetic and photoprotective pigments were analysed by HPLC. KEY RESULTS: Compared with glabrous-green leaves, both anthocyanic and pubescent leaves had greater dark-adapted PSII photochemical efficiency and net photosynthesis. In leaves possessing either anthocyanins or pubescence, the ratio of xanthophyll cycle components to total chlorophyll, and mid-day de-epoxidation state of the xanthophyll cycle were considerably smaller, than in glabrous-green leaves. These differences were more evident in pubescent leaves, probably indicating that trichomes were more effective in decreasing light stress than anthocyanins in the epidermis. CONCLUSIONS: Light screens, especially in the form of pubescence, decrease the risk of photoinhibition whilst allowing leaves to maintain a smaller content of xanthophyll cycle components and depend less on xanthophyll cycle energy dissipation. This combination of photoprotective features, i.e. decreased photon flux to the photosynthetic apparatus and lower xanthophyll cycle utilization rates may be particularly advantageous under stressful conditions.     

Enhanced Employment of the Xanthophyll Cycle and Thermal Energy Dissipation in Spinach Exposed to High Light and N Stress

The involvement of the xanthophyll cycle in photoprotection of N-deficient spinach (Spinacia oleracea L. cv Nobel) was investigated. Spinach plants were fertilized with 14 mM nitrate (control, high N) versus 0.5 mM (low N) fertilizer, and grown under both high- and low-light conditions. Plants were characterized from measurements of photosynthetic oxygen exchange and chlorophyll fluorescence, as well as carotenoid and cholorophyll analysis. Compared with the high-N plants, the low-N plants showed a lower capacity for photosynthesis and a lower chlorophyll content, as well as a lower rate of photosystem II photosynthetic electron transport and a corresponding increase in thermal energy dissipation activity measured as nonphotochemical fluorescence quenching. The low-N plants displayed a greater fraction of the total xanthophyll cycle pool as zeaxanthin and antheraxanthin at midday, and an increase in the ratio of xanthophyll cycle pigments to total chlorophyll. These results indicate that under N limitation both the light-collecting system and the photosynthetic rate decrease. However, the increased dissipation of excess energy shows that there is excess light absorbed at midday. We conclude that spinach responds to N limitation by a combination of decreased light collection and increased thermal dissipation involving the xanthophyll cycle.     

Photosynthesis and Photoprotection in Overwintering Plants

Abstract: Seasonal differences in the capacity of photosynthetic electron transport, leaf pigment composition, xanthophyll cycle characteristics and chlorophyll fluorescence emission were investigated in two biennial mesophytes ( Malva neglecta and Verbascum thapsus ) that grow in full sunlight, and in leaves/needles of sun and shade populations of several broad-leafed evergreens and conifers (Vinca minor, Euonymus kiautschovicus, Mahonia repens, Pseudotsuga menziesii [Douglas fir], and Pinus ponderosa). Both mesophytic species maintained or upregulated photosynthetic capacity in the winter and exhibited no upregulation of photoprotection. In contrast, photosynthetic capacity was downregulated in sun leaves/needles of V. minor, Douglas fir, and Ponderosa pine, and even in shade needles of Douglas fir. Interestingly, photosynthetic capacity was upregulated during the winter in shade leaves/needles of V. minor, Ponderosa pine and Euonymus kiautschovicus. Nocturnal retention of zeaxanthin and antheraxanthin, and their sustained engagement in a state primed for energy dissipation, were observed largely in the leaves/needles of sun-exposed evergreen species during winter. Factors that may contribute to these differing responses to winter stress, including chloroplast redox state, the relative levels of source and sink activity at the whole plant level, and apoplastic versus symplastic phloem loading, are discussed.     

Antioxidants and xanthophyll cycle-dependent energy dissipation in Cucurbita pepo L. and Vinca major L. acclimated to four growth PPFDs in the field

The acclimation of photochemistry, xanthophyll cycle-dependent energy dissipation, and antioxidants was characterized in leaves of Cucurbita pepo L. and Vinca major L. that developed under photosynthetic photon flux densities (PPFDs) ranging from deep shade to full sunlight in the field. The predominant acclimatory response of leaf pigment composition was an increase in the xanthophyll cycle pool size with increasing growth PPFD. In both species, the estimated rate of thermal energy dissipation at midday increased with increasing PPFD and midday levels of zeaxanthin and antheraxanthin per chlorophyll were closely correlated with the levels of non-photochemical fluorescence quenching under all growth PPFD regimes. However, at full sunlight there appeared to be considerably higher levels of xanthophyll cycle dependent energy dissipation in V. major compared with pumpkin while estimated rates of photochemistry exhibited the reverse trend. Leaf activities of the antioxidant enzymes ascorbate peroxidase and superoxide dismutase, as well as ascorbate content, increased with increasing growth PPFD in both plant species. Activities/contents were higher under 100% full sunlight and increased more strongly from intermediate growth PPFDs to 100% full sunlight in V. major than in C. pepo. These patterns of acclimation are similar to those exhibited by xanthophyll cycle-dependent energy dissipation. The patterns of acclimation of glutathione reductase are discussed in the context of the multiple roles for reduced glutathione. Catalase acclimated in a manner consistent with its role in scavenging H2O2 generated via photorespiration and/or mitochondrial respiration. Leaf -tocopherol did not exhibit growth PPFD-dependent trends.     

Photoprotective role of rhodoxanthin during cold acclimation in Cryptomeria japonica

To examine the role of rhodoxanthin in long‐term acclimation to low temperatures, we monitored seasonal changes in pigment composition, photosynthesis, chlorophyll fluorescence and the level of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) in needles of wild‐type and mutant forms of Cryptomeria japonica. In winter, rhodoxanthin accumulated in sun‐exposed needles of wild‐type plants, but not in those of the mutant. The level of chlorophyll decreased in both types of plant in winter. In contrast, the level of the xanthophyll cycle pool increased in both cases. The level of the pool in the mutant was twice that in the wild type in winter, on a Chl basis, even though the levels in both were similar in summer. The synthesis of rhodoxanthin might be triggered by photo‐inhibitory conditions, as suggested by the sustained elevated levels of zeaxanthin (Z) and antheraxanthin (A). In the wild type and the mutant, the quantum yield of CO₂ fixation (φ), the photosynthetic capacity, the photochemical efficiency of photosystem II (PSII), the photochemical quenching and the level of Rubisco in summer were similar. However, all these values for the wild type were higher than those for the mutant in winter. The non‐photochemical quenching (NPQ) in the mutant in winter increased rapidly even under low light conditions due to the high sustained levels of Z and A. In contrast, in the wild type, the conversion of Z via A to rhodoxanthin prevented the rapid increase in NPQ to maintain the relatively high level of φ. These findings suggest that rhodoxanthin might play an important photoprotective role in long‐term acclimation to cold. The dynamic regulation of the amount of rhodoxanthin relative to the level of the xanthophyll cycle pool might act to maintain an appropriate balance between light absorption, photosynthesis and the thermal dissipation of energy due to excess absorbed light in winter.     

Effects of changes in growth temperature on photosynthesis and carotenoid composition in Zea mays leaves

The effects of changes in growth temperature on photosynthesis and carotenoid composition were examined in Zea mays L. leaves of different age and different developmental history. The plants were first grown at sub-optimal temperature (14°C) until the full development of the third leaf. At that time, the mature third leaf and the immature fourth leaf had a low chlorophyll (Chl) content, a low Chl a/b ratio, a high carotenoid/Chl a+b ratio, a high xanthophyll/β-carotene ratio, and about 80% of the xanthophyll cycle pool (violaxanthin [V] + antheraxanthin [A] + zeaxanthin [Z]) was in the form of zeaxanthin and antheraxanthin. When the temperature was increased from 14°C to 24°C for three days, increased Chl synthesis, accompanied by an increase in the Chl a/b ratio, took place. The ratios of lutein, neoxanthin, and V+A+Z to Chl a+b decreased markedly, whereas no significant changes appeared in the β-carotene/Chl a+b ratio. Furthermore, there was a sharp decrease in the xanthophyll/β-carotene ratio and most of zeaxanthin was converted to violaxanthin in the xanthophyll cycle. The third leaf and the tip segment of the fourth leaf, both expanded at 14°C, showed little difference in their pigment contents. However, the rate of CO₂ assimilation of the tip segment of the fourth leaf was nearly twice that of the third leaf on the third day at 24°C, while the photosynthetic activity was similar in both leaves before the transfer to 24°C. During the warm period at 24°C, new leaf tissue (basal segment of the fourth leaf and part of a fifth leaf) was formed. On the third day at 24°C, the pigment content of 24°C-grown leaf tissue did not differ much from that of 14°C-grown leaf tissue with the exception that the total carotenoid content was lower in the former as compared to the latter, mainly because of a lower V+A+Z content. The rate of CO₂ assimilation of 24°C-grown leaf tissue was comparable to that of the tip segment of the fourth leaf. Regardless of which leaf tissue is considered, reducing the temperature from 24°C to 14°C for 5 days slightly affected the pigment content, but violaxanthin was largely converted to zeaxanthin and antheraxanthin in the xanthophyll cycle. The results indicate that compared to old leaf tissue of mature leaves, physiologically younger leaf tissue of immature leaves is much more able to recover from depressions in the photosynthetic activity induced by growth at sub-optimal temperature when the plants experience optimal growth temperatures, but that factors other than the pigment content must determine this capability.     

Positive correlation between levels of retained zeaxanthin + antheraxanthin and degree of photoinhibition in shade leaves of Schefflera arboricola (Hayata) Merrill

Attached intact leaves of Schefflera arboricola grown at three different photon flux densities (PFDs) were subjected to 24-h exposures to a high PFD and subsequent recovery at a low PFD. While sun leaves showed virtually no sustained effects on photosystem II (PSII), shade-grown leaves exhibited pronounced photoinhibition of PSII that required several days at low PFD to recover. Upon transfer to high PFD, levels of nonphotochemical quenching in PSII as well as levels of zeaxanthin were initially low in shade leaves but continued to increase gradually during the 24-h exposure. The xanthophyll cycle pool size rose gradually during and also subsequent to the photoinhibitory treatment in shade leaves. Upon return to low PFD, a marked and extremely long-lasting retention of zeaxanthin and antheraxanthin was observed in shade but not sun leaves. During recovery, changes in the conversion state of the xanthophyll cycle therefore closely mirrored the slow increases in PSII efficiency. This novel report of a close association between zeaxanthin retention and lasting PSII depressions in these shade leaves clearly suggests a role for zeaxanthin in photoinhibition of shade leaves. In addition, there was a decrease in β-carotene levels, some decrease in chlorophyll, but no change in lutein and neoxanthin (all per leaf area) in the shade leaves during and subsequent to the photoinhibitory treatment. These data may be consistent with a degradation of a portion of core complexes but not of peripheral light-harvesting complexes. A possible conversion of β-carotene to form additional zeaxanthin is discussed. Received: 24 October 1997 / Accepted: 12 November 1997     

Leaf orientation and the response of the xanthophyll cycle to incident light

Summary Leaves from two species, Euonymus kiautschovicus and Arctostaphylos uva-ursi, with a variety of different orientations and exposures, were examined in the field with regard to the xanthophyll cycle (the interconversion of three carotenoids in the chloroplast thylakoid membranes). East-, south-, and west-facing leaves of E. kiautschovicus were sampled throughout the day and all exhibited a pronounced and progressive conversion of violaxanthin to zeaxanthin, followed by a reconversion of zeaxanthin to violaxanthin later in the day. Maximal levels of zeaxanthin and minimal levels of violaxanthin were observed at the time when each leaf (orientation) received the maximum incident light, which was in the morning in east-facing, midday in southfacing, and in the afternoon in west-facing leaves. A very slight degree of hysteresis in the removal of zeaxanthin compared to its formation with regard to incident light was observed. Leaves with a broader range of orientations were sampled from A. uva-ursi prior to sunrise and at midday. All of the examined pigments (carotenoids and chlorophylls) increased somewhat per unit leaf area with increasing total daily photon receipt. The sum of the carotenoids involved in the xanthophyll cycle, violaxanthin + antheraxanthin + zeaxanthin, increased more strongly with increasing growth light than any other pigment. In addition, the amounts of zeaxanthin present at midday also increased markedly with increasing total daily photon receipt. The percentage of the xanthophyll cycle that was converted to zeaxanthin (and antheraxanthin) at peak irradiance was very large (approximately 80%) in the leaves of both E. kiautschovicus and A. uva-ursi. The daily changes in the components of the xanthophyll cycle that paralleled the daily changes in incident light in the leaves of E. kiautschovicus, and the increasing levels of the xanthophyll cycle components with total daily photon receipt in the leaves of A. uva-ursi, are both consistent with the involvement of zeaxanthin (i.e. the xanthophyll cycle) in the photoprotection of the photosynthetic apparatus against damage due to excessive light.Abbreviations A antheraxanthin - EPS epoxidation state of the xanthophyll cycle=(V+0.5A)/(V+A+Z) - PFD photon flux density (400–700 nm) - PFD_i photon flux density incident upon the upper leaf surface - T_air air temperature - T_L leaf temperature - V violaxanthin - Z zeaxanthin