Mesophyll versus epidermal anthocyanins as potential in vivo antioxidants: evidence linking the putative antioxidant role to the proximity of oxy-radical source

The hypothesis that anthocyanins in red leaves may be potential in vivo antioxidants whose efficiency is linked to their proximity with the oxy-radical source was tested. Advantage was taken of intra-individual and intra-species variations in the anthocyanic trait and green and red leaves on the same individuals or leaves of green and red phenotypes were compared for the extent of PSII damage by reactive oxygen species generated by methyl viologen treatment in the light. Two species possessing anthocyanins in the mesophyll (Cistus creticus and Photinia x fraseri) and two in the epidermis (Rosa sp. and Ricinus communis) were used, while red actinic light (which is not absorbed by anthocyanins) allowed discrimination between an indirect sunscreen and a direct antioxidant function. Red leaves whose anthocyanins were located in the mesophyll were more resistant to methyl viologen treatment than their green counterparts. In one of these species (Cistus creticus), where anthocyanins are induced in some individuals within the natural population after bright cool days in winter, both green and future-red morphs displayed the same sensitivity to methyl viologen before anthocyanin induction. Immediately after reddening, however, resistance to methyl viologen was considerably increased in the red morphs. By contrast, red leaves whose anthocyanins were restricted to epidermal cells were more sensitive to the herbicide. Total leaf phenolic levels in green/red pairs were similar. The results indicate that vacuolar anthocyanins may be an effective in vivo target for oxy-radicals, provided that the oxy-radical source and the anthocyanic detoxifying sink are in close vicinity.     

Xanthophyll cycle pigment and antioxidant profiles of winter-red (anthocyanic) and winter-green (acyanic) angiosperm evergreen species

Leaves of many angiosperm evergreen species change colour from green to red during winter, corresponding with the synthesis of anthocyanin pigments. The ecophysiological function of winter colour change (if any), and why it occurs in some species and not others, are not yet understood. It was hypothesized that anthocyanins play a compensatory photoprotective role in species with limited capacity for energy dissipation. Seasonal xanthophyll pigment content, chlorophyll fluorescence, leaf nitrogen, and low molecular weight antioxidants (LMWA) of five winter-red and five winter-green angiosperm evergreen species were compared. Our results showed no difference in seasonal xanthophyll pigment content (V+A+Z g(-1) leaf dry mass) or LMWA between winter-red and winter-green species, indicating red-leafed species are not deficient in their capacity for non-photochemical energy dissipation via these mechanisms. Winter-red and winter-green species also did not differ in percentage leaf nitrogen, corroborating previous studies showing no difference in seasonal photosynthesis under saturating irradiance. Consistent with a photoprotective function of anthocyanin, winter-red species had significantly lower xanthophyll content per unit chlorophyll and less sustained photoinhibition than winter-green species (i.e. higher pre-dawn F(v)/F(m) and a lower proportion of de-epoxidized xanthophylls retained overnight). Red-leafed species also maintained a higher maximum quantum yield efficiency of PSII at midday (F'(v)/F'(m)) during winter, and showed characteristics of shade acclimation (positive correlation between anthocyanin and chlorophyll content, and negative correlation with chlorophyll a/b). These results suggest that the capacity for photon energy dissipation (photochemical and non-photochemical) is not limited in red-leafed species, and that anthocyanins more likely function as an alternative photoprotective strategy to increased VAZ/Chl during winter.     

The winter-red-leaf syndrome in Pistacia lentiscus: evidence that the anthocyanic phenotype suffers from nitrogen deficiency, low carboxylation efficiency and high risk of photoinhibition

Recent evidence indicates that winter-red leaf phenotypes in the mastic tree (Pistacia lentiscus) are more vulnerable to chronic photoinhibition during the cold season relative to winter-green phenotypes occurring in the same high light environment. This was judged by limitations in the maximum quantum yield of photosystem II (PSII), found in previous studies. In this investigation, we asked whether corresponding limitations in leaf gas exchange and carboxylation reactions could also be manifested. During the cold (“red”) season, net CO₂ assimilation rates (A) and stomatal conductances (g_s) in the red phenotype were considerably lower than in the green phenotype, while leaf internal CO₂ concentration (Ci) was higher. The differences were abolished in the “green” period of the year, the dry summer included. Analysis of A versus Ci curves indicated that CO₂ assimilation during winter in the red phenotype was limited by Rubisco content and/or activity rather than stomatal conductance. Leaf nitrogen levels in the red phenotype were considerably lower during the red-leaf period. Consequently, we suggest that the inherently low leaf nitrogen levels are linked to the low net photosynthetic rates of the red plants through a decrease in Rubisco content. Accordingly, the reduced capacity of the carboxylation reactions to act as photosynthetic electron sinks may explain the corresponding loss of PSII photon trapping efficiency, which cannot be fully alleviated by the screening effect of the accumulated anthocyanins.     

Role of red carotenoids in photoprotection during winter acclimation in Buxus sempervirens leaves

The red leaf coloration of several plant species during autumn and winter is due to the synthesis of phenolic compounds such as anthocyanins or red carotenoids. The latter occur very rarely and are non-ubiquitous and taxonomically restricted compounds. The present study shows that the leaves of common box ( Buxus sempervirens L.) accumulate red carotenoids (eschscholtzxanthin, monoanhydroeschscholtzxanthin, anhydroeschscholtzxanthin) as a response to photoinhibitory conditions during winter acclimation. These compounds are produced in a coordinated manner with the operation of other photoprotective systems: accumulation and sustained deepoxidation of VAZ pigments with a concomitant decrease in maximal photochemical efficiency, accumulation of alpha-tocopherol and a gradual decrease on chlorophyll content. All these processes were reversed when the photosynthetic tissues were transferred from photoinhibitory winter conditions to room temperature for 9 days. Buxus leaves showed a large degree of phenotype variation in the degree of reddening, ranging from green to orange. The differences in colour pattern were mainly due to differences in the accumulation of red carotenoids and xanthophyll esters. Red pigments were mainly anhydroeschscholtzxanthin and esters of eschscholtzxanthin. Conversely to fruit or petal chromoplasts, the plastids of red leaves in this species are not the terminal differentiated state but are able to redifferentiate again to chloroplasts. Their photoprotective role during winter as a light screen system or as antioxidants, in a similar way to other red pigments, and their implications on the wide ecological tolerance of this evergreen species are discussed.     

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.     

Functional role of red (retro)-carotenoids as passive light filters in the leaves of Buxus sempervirens L.: increased protection of photosynthetic tissues?

Red (retro)-carotenoids accumulate in chloroplasts of Buxus sempervirens leaves during the process of winter leaf acclimation. As a result of their irregular presence, different leaf colour phenotypes can be found simultaneously in the same location. Five different colour phenotypes (green, brown, red, orange, and yellow), with a distinct pattern of pigment distribution and concentration, have been characterized. Leaf reddening due to the presence of anthocyanins or carotenoids, is a process frequently observed in plant species under photoinhibitory situations. Two main hypotheses have been proposed to explain the function of such colour change: antioxidative protection exerted by red-coloured molecules, and green light filtering. The potential photoprotective role of red (retro-) carotenoids as light filters was tested in Buxus sempervirens leaves. In shade leaves of this species the upper (adaxial) mesophyll of the lamina was replaced by the equivalent upper part of a different colour phenotype. These hybrid leaves were exposed to a photoinhibitory treatment in order to compare the photoprotective effect exerted by adaxial parts of phenotypes with a different proportion of red (retro)-carotenoids in the lower mesophyll of a shade leaf. The results indicated that the presence of red (retro)-carotenoids in the upper mesophyll did not increase photoprotection of the lower mesophyll when compared with chlorophyll, and the best protection was achieved by an upper green layer. This was due to the fact that the extent of photoinhibition was proportional to the amount of red light transmitted by the upper mesophyll and/or to the chlorophyll pool located above. These results do not exclude a protective function of carotenoids in the upper leaf layer, but imply that, at least under the conditions of this experiment, the accumulation of red pigments in the outer leaf layers does not increase photoprotection in the lower mesophyll.     

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.     

Epidermal coumaroyl anthocyanins protect sweet basil against excess light stress: multiple consequences of light attenuation

The putative photoprotective role of foliar anthocyanins continues to attract heated debate. Strikingly different experimental set‐ups coupled with a poor knowledge of anthocyanin identity have likely contributed to such disparate opinions. Here, the photosynthetic responses to 30 or 100% solar irradiance were compared in two cultivars of basil, the green‐leafed Tigullio (TG) and the purple‐leafed Red Rubin (RR). Coumaroyl anthocyanins in RR leaf epidermis significantly mitigated the effects of high light stress. In full sunlight, RR leaves displayed several shade‐plant traits; they transferred less energy than did TG to photosystem II (PSII), and non‐photochemical quenching was lower. The higher xanthophyll cycle activity in TG was insufficient to prevent inactivation of PSII in full sunlight. However, TG was the more efficient in the shade; RR was far less able to accommodate a large change in irradiance. Investment of carbon to phenylpropanoid biosynthesis was more in RR than in TG in the shade, and was either greatly enhanced in TG or varied little in RR because of high sunlight. The metabolic cost of photoprotection was lower whereas light‐induced increase in biomass production was higher in RR than in TG, thus making purple basil the more light tolerant. Purple basil appears indeed to display the conservative resource‐use strategy usually observed in highly stress tolerant species. We conclude that the presence of epidermal coumaroyl anthocyanins confers protective benefits under high light, but it is associated with a reduced plasticity to accommodate changing light fluxes as compared with green leaves.     

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.     

Strength of winter leaf redness as an indicator of stress vulnerable individuals in Pistacia lentiscus

Leaf anthocyanins are believed to afford protection against photoinhibition, yet the dependence of protection on actual anthocyanin concentrations has not been investigated. To that aim, non-invasive optical methods (spectral reflectance and chlorophyll fluorescence) were used to assess the levels of anthocyanins and chlorophylls as well as photosystem II photochemical efficiency in hundreds of leaves from the mastic tree (Pistacia lentiscus), which displays in the field a continuum of leaf tints during winter from fully green to fully red. Contrary to expectations based on the photoprotective hypothesis, the strength of leaf redness was positively correlated to the extent of mid-winter chronic photoinhibition and negatively correlated to leaf chlorophyll contents. Hence, a photoprotective role for anthocyanins is not substantiated. Instead, we argue that winter leaf redness may be used reliably, quickly and non-invasively to locate vulnerable individuals in the field.     

Red 'Anjou' pear has a higher photoprotective capacity than green 'Anjou'

Photoprotective function of anthocyanins along with xanthophyll cycle and antioxidant system in fruit peel was investigated in red 'Anjou' vs green 'Anjou' pear (Pyrus communis) during fruit development and in response to short-term exposure to high light. The sun-exposed peel of red 'Anjou' had higher maximum quantum yield of photosystem II (F(V)/F(M)) than that of green 'Anjou' and both the sun-exposed peel and the shaded peel of red 'Anjou' had smaller decreases in F(V)/F(M) after 2-h high light (photon flux density of 1500 mumol m(-2) s(-1)) treatment than those of green 'Anjou'. At the middle and late developmental stages, the xanthophyll cycle pool size on a chlorophyll basis, the activity of superoxide dismutase, ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR), dehydroascorbate reductase (DHAR) and glutathione reductase (GR) and the level of reduced ascorbate and total ascorbate pool in the sun-exposed peel were either the same or lower in red 'Anjou' than in green 'Anjou', whereas the xanthophyll cycle pool size on a chlorophyll basis and the activity of APX, catalase, MDAR, DHAR and GR in the shaded peel were higher in red 'Anjou' than in green 'Anjou'. It is concluded that red 'Anjou' has a higher photoprotective capacity in both the sun-exposed peel and the shaded peel than green 'Anjou'. While the higher anthocyanin concentration along with the larger xanthophyll cycle pool size and the higher activity of some antioxidant enzymes may collectively contribute to the higher photoprotective capacity in the shaded peel of red 'Anjou', the higher photoprotective capacity in the sun-exposed peel of red 'Anjou' is mainly attributed to its higher anthocyanin concentration.     

Functional role of anthocyanins in high-light winter leaves of the evergreen herb Galax urceolata

High-light leaves of the evergreen herb Galax urceolata exhibit a striking color change from green to red during winter months due to anthocyanin synthesis in outermost mesophyll cells. Here we investigate three possible functions of this color change. To test the hypothesis that anthocyanins function as light attenuators, maximum photosystem II efficiency (F(v)/F(m)) of red and green leaves was measured during and after exposure to wavelengths either strongly or poorly absorbed by anthocyanin. To determine whether anthocyanins elevate radical-scavenging capacity, antioxidant activity of red and green leaves was assessed using the alpha,alpha-diphenyl-beta-picrylhydrazyl assay. Nonstructural carbohydrate levels were analyzed to test the hypothesis that anthocyanins function as a carbon sink. Declines in F(v)/F(m) under white and green light were significantly greater for green than red leaves, but were comparable under red light. Anthocyanin content positively correlated with antioxidant activity. Although levels of anthocyanins did not appear to be related to nonstructural carbohydrate concentration, high levels of sugars may be necessary for their photo-induction. Results suggest that anthocyanins function as light attenuators and may also contribute to the antioxidant pool in winter leaves.     

Seasonal differences in xanthophyll cycle characteristics and antioxidants in Mahonia repens growing in different light environments

We investigated differences between summer and winter in photosynthesis, xanthophyll cycle-dependent energy dissipation, and antioxidant systems in populations of Mahonia repens (Lindley) Don growing in the eastern foothills of the Colorado Rocky Mountains in deep shade, full exposure, and under a single-layered canopy of Pinus ponderosa (partially shaded). In summer, increasing growth irradiance (from deep shade to partial shade to full exposure) was associated with increased xanthophyll cycle-dependent energy dissipation in PSII and an increased capacity to detoxify reactive reduced oxygen species, as measured by increases in the activities of ascorbate peroxidase, superoxide scavenging, glutathione reductase, and monodehydroascorbate reductase, as well as increases in leaf ascorbate and glutathione content. Leaves of exposed and partially shaded plants exhibited decreased capacities for photosynthetic O₂ evolution in winter compared to summer, while in the deeply shaded plants this parameter did not differ seasonally. Seasonal differences in the levels of antioxidants generally exhibited an inverse response to photosynthesis, being higher in winter compared to summer in the exposed and partially shaded populations, but remaining unchanged in the deeply shaded population. In addition, total pool size and conversion state of the xanthophyll cycle were higher in winter than in summer in all populations. These trends suggest that both xanthophyll cycle-dependent energy dissipation in PSII and the capacity to detoxify reactive reduced oxygen species responded to the level of excess light absorption. Received: 23 October 1997 / Accepted: 23 March 1998     

Seasonal reversibility of acclimation to irradiance in leaves of common box (Buxus sempervirens L.) in a deciduous forest

Understorey shade plants are seasonally exposed to dramatic changes in light conditions in deciduous forests related with the dynamics of the overstorey leaf phenology. These transitions are commonly followed by changes in herb plant communities, but shade-tolerant evergreen species must be able to adapt to changing light conditions. In this work we checked the photoprotective responses of evergreen species to acclimate to the shady summer environment and reversibly de-acclimate to a more illuminated environment after leaf fall on deciduous overstoreys. For that purpose we have followed the process of light acclimation in leaves of common box (Buxus sempervirens) during the winter to spring transition, which decrease irradiance in the understorey, and conversely during the transition from summer to autumn. Four parameters indicative of the structure and degree of acclimation of the photosynthetic apparatus have been studied: chlorophyll a/b ratio which is supposed to be inversely proportional to the antenna size, α/β-carotene which increases in shade acclimated leaves and the pools of α-tocopherol and xanthophyll cycle pigments (VAZ) which are two of the main photoprotection mechanisms in plants. Among these parameters, chlorophyll a/b ratio and VAZ pool responded finely to changes in irradiance indicating that modifications in the light harvesting size and photoprotective capacity contribute to the continuous acclimation and de-acclimation of long-lived evergreen leaves.     

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 (R_RED: R_GREEN), 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 (Φ_PSII) 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.     

Variations in light energy dissipation in <Emphasis Type="Italic">Woodfordia fruticosa</Emphasis> leaves during expansion

Young leaves of tropical trees frequently appear red in color, with the redness disappearing as the leaves mature. During leaf expansion, plants may employ photoprotective mechanisms to cope with high light intensities; however, the variations in anthocyanin contents, nonphotochemical quenching (NPQ), and photorespiration during leaf expansion are poorly understood. Here, we investigated pigment contents, gas exchange, and chlorophyll (Chl) fluorescence in Woodfordia fruticosa leaves during their expansion. Young red leaves had significantly lower Chl content than that of expanding or mature leaves, but they accumulated significantly higher anthocyanins and dissipated more excited light energy through NPQ. As the leaves matured, net photosynthetic rate, total electron flow through PSII, and electron flow for ribulose-1,5-bisphosphate oxygenation gradually increased. Our results provided evidence that photorespiration is of fundamental importance in regulating the photosynthetic electron flow and CO₂ assimilation during leaf expansion.     

Photosynthetic activity of variegated leaves of Coleus × hybridus hort. cultivars characterised by chlorophyll fluorescence techniques

Different pigments often occur together and affect photosynthetic characteristics of the respective leaf portions. In this study, photosynthetic activity in variegated leaves of five cultivars of the ornamental and medicinal plant, Coleus × hybridus hort., was estimated by image analysis and point data measurements of major chlorophyll (Chl) fluorescence parameters and related to the amount of photosynthetic pigments measured with a Chl meter or spectrophotometrically in leaf extracts. Significant differences in Chl and carotenoid (Car) contents were noticed among differentially pigmented sectors of a leaf and among the cultivars. Although the higher Chl concentration was noticed in purple parts compared to green parts of the leaves, the values of minimal and maximal fluorescence yield at the dark- and light-adapted state (F₀, F_m, F₀', F_m', respectively) were a little lower than those in the green sectors, indicating photoprotective effects provided by anthocyanins and Car, more abundant in the red parts. The lowest Chl and Car content was detected in creamy-yellow and pink sectors and this contributed to low F₀, F_m, and F_m', maximal quantum yield of PSII photochemistry, and nonphotochemical and photochemical quenching but high PSII maximum efficiency and effective quantum yield of PSII photochemistry. Both methods of Chl fluorescence analysis revealed heterogeneity in capture, transfer, and dissipation of excitation energy but Chl fluorescence imaging was more suitable in examining very narrow pigmented leaf areas.     

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.     

Why leaves turn red in autumn. The role of anthocyanins in senescing leaves of red-osier dogwood

Why the leaves of many woody species accumulate anthocyanins prior to being shed has long puzzled biologists because it is unclear what effects anthocyanins may have on leaf function. Here, we provide evidence for red-osier dogwood (Cornus stolonifera) that anthocyanins form a pigment layer in the palisade mesophyll layer that decreases light capture by chloroplasts. Measurements of leaf absorbance demonstrated that red-senescing leaves absorbed more light of blue-green to orange wavelengths (495-644 nm) compared with yellow-senescing leaves. Using chlorophyll a fluorescence measurements, we observed that maximum photosystem II (PSII) photon yield of red-senescing leaves recovered from a high-light stress treatment, whereas yellow-senescing leaves failed to recover after 6 h of dark adaptation, which suggests photo-oxidative damage. Because no differences were observed in light response curves of effective PSII photon yield for red- and yellow-senescing leaves, differences between red- and yellow-senescing cannot be explained by differences in the capacities for photochemical and non-photochemical light energy dissipation. A role of anthocyanins as screening pigments was explored further by measuring the responses PSII photon yield to blue light, which is preferentially absorbed by anthocyanins, versus red light, which is poorly absorbed. We found that dark-adapted PSII photon yield of red-senescing leaves recovered rapidly following illumination with blue light. However, red light induced a similar, prolonged decrease in PSII photon yield in both red- and yellow-senescing leaves. We suggest that optical masking of chlorophyll by anthocyanins reduces risk of photo-oxidative damage to leaf cells as they senesce, which otherwise may lower the efficiency of nutrient retrieval from senescing autumn leaves.     

Hyponastic leaf growth decreases the photoprotective demand, prevents damage to photosystem II and delays leaf senescence in Salvia broussonetii plants

The influence of leaf angle on the response of plants to high light was studied in Salvia broussonetii, a species endemic of the Canary Islands that shows hyponastic leaf growth. The response of vertical, naturally oriented leaves was compared with that of horizontal, artificially held leaves for 1, 13, 24 and 29 days in terms of photoinhibition [efficiency of photosystem II (PSII)], photoprotection (by the xanthophyll cycle, alpha-tocopherol and beta-carotene) and progression of leaf senescence. Vertical leaves not only showed a decreased photoprotective demand compared with horizontal leaves but also kept the maximum efficiency of PSII (F(v)/F(m) ratio) constant throughout the experiment, thus reflecting the capacity of naturally oriented leaves to avoid photooxidative stress in the field. By contrast, horizontal leaves, which were exposed to higher light intensities, showed a higher photoprotective demand (reflected by a higher de-epoxidation of the xanthophyll cycle, carotenoid losses and increases in alpha-tocopherol), damage to PSII (as indicated by decreases in the F(v)/F(m) ratio) and accelerated leaf senescence, which was associated with cell death after 24 days of high light exposure. It is concluded that hyponastic leaf growth prevents photoinhibition and decreases the photoprotective demand of leaves by reducing the incident light, which helps maintaining leaf vigor and delaying the progression of leaf senescence in S. broussonetii plants. Hyponastic leaf growth is therefore one of the first photoprotection mechanisms activated in this species to avoid the negative impact of high-light stress in the field.