Photoprotective implications of leaf variegation in E. dens-canis L. and P. officinalis L

Variegated leaves occur rarely in nature, but there are some species, primarily in the forest understory, that possess this characteristic. We recently studied two variegated plants: Erytronium dens-canis L., which is characterised by a pattern of red patches and Pulmonaria officinalis L., with light green spots. These non-green areas could attenuate light reaching mesophyll cells with respect to green sections. The aim of the study was to verify whether such red and light green parts are more photoprotected than green ones and if this trait could be of adaptive value. Red patches in E. dens-canis were due to a single layer of red cells in the upper parenchyma, which accumulated anthocyanins. Light green spots in P. officinalis were caused by the presence of loosely arranged cells instead of a well-established layer of packed cells in the palisade parenchyma. Chlorophyll fluorescence imaging was performed under light treatment, showing a greater decrease of photochemical efficiency in red and light green patches than in green sections. Differences in the extent of photochemical efficiency among patches were not attributable to different activation of the xanthophyll cycle. These observations failed to confirm our initial hypothesis, but they questioned the physiological reason for this higher sensitivity in red and light green patches of photosynthetic tissues. Chlorophyll fluorescence imaging was therefore performed in the field. The same pattern of photochemical efficiency was maintained only in E. dens-canis. The current results demonstrate that in both species the benefits of variegation, if any, are different from enhanced photosynthetic performance.     

The applicability of empirical vegetation indices for determining leaf chlorophyll content over different leaf and canopy structures

Retrieving leaf chlorophyll content at a range of spatio-temporal scales is central to monitoring vegetation productivity, identifying physiological stress and managing biological resources. However, estimating leaf chlorophyll over broad spatial extents using ground-based traditional methods is time and resource heavy. Satellite-derived spectral vegetation indices (VIs) are commonly used to estimate leaf chlorophyll content, however they are often developed and tested on broadleaf species. Relatively little research has assessed VIs for different leaf structures, particularly needle leaves which represent a large component of boreal forest and significant global ecosystems. This study tested the performance of 47 published VIs for estimating foliar chlorophyll content from different leaf and canopy structures (broadleaf and needle). Coniferous and deciduous sites were selected in Ontario, Canada, representing different dominant vegetation species (Picea mariana and Acer saccharum) and a variety of canopy structures. Leaf reflectance data was collected using an ASD Fieldspec Pro spectroradiometer (400–2500 nm) for over 300 leaf samples. Canopy reflectance data was acquired from the medium resolution imaging spectrometer (MERIS). At the canopy level, with both leaf types combined, the DD-index showed the strongest relationship with leaf chlorophyll (R² = 0.78; RMSE = 3.56 μg/cm²), despite differences in leaf structure. For needleleaf trees alone the relationship with the top VI was weaker (D_[red], R² = 0.71; RMSE = 2.32 μg/cm²). A sensitivity study using simulated VIs from physically-modelled leaf (PROSPECT) and canopy (4-Scale) reflectance was performed in order to further investigate these results and assess the impacts of different background types and leaf area index on the VIs’ performance. At the leaf level, the MNDVI8 index showed a strong linearity to changing chlorophyll and negligible difference to leaf structure/type. At canopy level, the best performing VIs were relatively consistent where LAI ≥ 4, but responded strongly to differences in background at low canopy coverage (LAI = 2). This research provides comprehensive assessments for the use of spectral indices in retrieval of spatially-continuous leaf chlorophyll content at the leaf (MTCI: R² = 0.72; p < 0.001) and canopy (DD: R² = 0.78; p < 0.001) level for resource management over different spatial and temporal scales.     

Hyperspectral indices for estimating leaf biochemical properties in temperate deciduous forests: Comparison of simulated and measured reflectance data sets

This study aimed at finding efficient hyperspectral indices for estimating three leaf biochemical parameters: chlorophyll content (CHL, μg cm⁻²), leaf water thickness (EWT, g cm⁻²), and leaf mass per area (LMA, g cm⁻²) in typical temperate deciduous forests. These parameters are required by most biogeochemical models that describe ecosystem functions. We have identified the most efficient hyperspectral indices (both the index types and the wavelength domains) based on both a simulated data set (produced with the calibrated leaf reflectance model PROSPECT) and with data sets (I, II, and III) from measurement of field-collected leaves. Results indicated that CHL, EWT, and LMA can be estimated with high precision using a two-waveband vegetation index (Double Deference index, DDn) for all parameters, with an overall root mean square error (RMSE) of 6.87 μg cm⁻² for CHL, 0.0011 g cm⁻² for EWT, and 0.0015 g cm⁻² for LMA. The best overall indices for temperate deciduous forests were DDn (715, 185) for CHL, DDn (1530, 525) for EWT, and DDn (1235, 25) for LMA, although these indices were not necessarily the best for every specific data set (especially for the simulated data set). Moreover, discrepancies were obvious when the identified indices were applied to different data sets. Even if the wavelengths of calibrated indices have been accurately determined through the simulated data set, the regressions between the indices and the biochemical parameters must be calibrated with field-based measurements. The indices identified in this study are applicable to various species (data set III), various phenological stages and locations (data set I), and various leaf anatomies (data set II) and may therefore be widely applicable for temperate deciduous forests and possibly for other plant communities.     

The effects of leaf size,leaf habit,and leaf form on leaf/stem relationships in plant twigs of temperate woody species

Question: Do thick‐twigged/large‐leaf species have an advantage in leaf display over their counterparts, and what are the effects of leaf habit and leaf form on the leaf‐stem relationship in plant twigs of temperature broadleaf woody species? Location: Gongga Mountain, southwest China. Methods: (1) We investigated stem cross‐sectional area and stem mass, leaf area and leaf/lamina mass of plant twigs (terminal branches of current‐year shoots) of 89 species belonging to 55 genera in 31 families. (2) Data were analyzed to determine leaf‐stem scaling relationships using both the Model type II regression method and the phylogenetically independent comparative (PIC) method. Results: (1) Significant, positive allometric relationships were found between twig cross‐sectional area and total leaf area supported by the twig, and between the cross‐sectional area and individual leaf area, suggesting that species with large leaves and thick twigs could support a disproportionately greater leaf area for a given twig cross‐sectional area. (2) However, the scaling relationships between twig stem mass and total leaf area and between stem mass and total lamina mass were approximately isometric, which indicates that the efficiency of deploying leaf area and lamina mass was independent of leaf size and twig size. The results of PIC were consistent with these correlations. (3) The evergreen species were usually smaller in total leaf area for a given twig stem investment in terms of both cross‐sectional area and stem mass, compared to deciduous species. Leaf mass per area (LMA) was negatively associated with the stem efficiency in deploying leaf area. (4) Compound leaf species could usually support a larger leaf area for a given twig stem mass and were usually larger in both leaf size and twig size than simple leaf species. Conclusions: Generally, thick‐twigged/large‐leaf species do not have an advantage over their counterparts in deploying photosynthetic compartments for a given twig stem investment. Leaf habit and leaf form types can modify leaf‐stem scaling relationships, possibly because of contrasting leaf properties. The leaf size‐twig size spectrum is related to the LMA‐leaf life span dimension of plant life history strategies.     

Characteristics of Encelia species differing in leaf reflectance and transpiration rate under common garden conditions

Summary The performance of coastal and desert species of Encelia (Asteraceae) were evaluated through common garden growth observations. The obectives of the study were to evaluate the roles of leaf features, thought to be of adaptive value (increased leaf reflectance and/or transpirational cooling), on plant growth in the hot, arid, desert garden versus their impact on growth under cooler, relatively more moist coastal garden conditions. E. californica native to the coast of southern California and E. farinosa, and E. frutescens, interior desert species, were grown in common gardens at coastal (Irvine, California) and interior (Phoenix, Arizona) sites under both irrigated and natural conditions. Although all species survived in both gardens during the two and a half year study period, there were large differences in their sizes. In the desert garden, leaf conductance and leaf water potential were both lower than at the coastal site. E. californica shrubs were leafless much of the time under natural conditions in the desert garden and had the smallest size there as well. Under natural conditions, E. farinosa, with its reflective leaf surface, was able to maintain lower leaf temperatures and attained a large size than the other two species in the desert garden. The green-leaved species (E. californica and E. frutescens) were not able to maintain leaves into the drought periods in the desert garden, with the exception of the irrigated E. frutescens which did maintain its leaf area if provided with supplemental watering to maintain transpirational leaf cooling. In the coastal garden, all species survived and there were few clear differences in the physiological characteristics among the three species. E. californica, the coastal native, attained a larger size in the coastal garden when compared with either of the two desert species.     

The development of the activity of the chloroplastic and cytosolic isoenzymes of phosphoglycerate kinase during barley leaf ontogenesis

The catalytic activities of the chloroplastic and cytosolic isoenzymes of phosphoglycerate kinase (PGK; EC 2.7.2.3) have been followed during the development of the first leaf of barley (Hordeum vulgare L.) grown for 7 d in darkness followed by transfer to continuous illumination. The investigation has included both the study of a standard leaf section, measured from the leaf tip, over the whole life of the leaf and the study of serial sections of leaf, measured from the leaf base, at a standard sampling time. The results of both approaches were fully compatible. As the catalytic activity of each isoenzyme in the standard assay is directly proportional to the amount of isoenzyme protein present, the catalytic activities may be interpreted wholly in terms of enzyme synthesis and degradation. Both isoenzymes are synthesized in darkness and in etiolated barley are present at a ratio of about 2674 for the cytosolic to chloroplastic isoenzymes. Illumination results in a fivefold or greater increase in chloroplast PGK over a number of days with little change of the cytosolic isoenzyme, resulting in an eventual ratio of cytosolic to chloroplastic isoenzymes approaching the green-leaf value of about 991. Prior to any detectable onset of senescence a 15-fold increase in cytosolic isoenzyme commenced while the amount of chloroplast PGK remained constant. It is suggested that the increased cytosolic PGK may be involved in the export of carbohydrate reserves (starch) prior to leaf senescence. Both isoenzymes subsequently decline in parallel to total protein and chlorophyll in the course of senescence.Abbreviations DHAP reductase dihydroxyacetone-phosphate reductase - GS glutamine synthetase - LHCP light-harvesting chlorophyll-a/b-binding protein - PGK phosphoglycerate kinase - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase This work was supported by the Science and Engineering Research Council (grant no. GR/E54504).     

Constancy of organellar genome copy numbers during leaf development and senescence in higher plants

In higher plants, plastid and mitochondrial genomes occur at high copy numbers per cell. Several recent publications have suggested that, in higher plants like Arabidopsis and maize, chloroplast DNA is virtually absent in mature and old leaves. This conclusion was mainly based on DAPI staining of isolated chloroplasts. If correct, the finding that chloroplasts in mature leaves lack DNA would change dramatically our understanding of gene expression, mRNA stability and protein stability in chloroplasts. In view of the wide implications that the disposal of chloroplast DNA during leaf development would have, we have reinvestigated the age dependency of genome copy numbers in chloroplasts and, in addition, tested for possible changes in mitochondrial genome copy number during plant development. Analyzing chloroplast and mitochondrial DNA amounts in Arabidopsis and tobacco plants, we find that organellar genome copy numbers remain remarkably constant during leaf development and are present in essentially unchanged numbers even in the senescing leaves. We conclude that, during leaf development, organellar gene expression in higher plants is not significantly regulated at the level of genome copy number and we discuss possible explanations for the failure to detect DNA in isolated chloroplasts stained with DAPI.     

The role of carbon dioxide and oxygen in determining chlorophyll fluorescence quenching during leaf development

Chlorophyll fluorescence emission at 680 nm (F680) and the rate of CO₂ fixation were measured simultaneously in sections along the length of wheat and maize leaves. These leaves possess a basal meristem and show a gradation in development towards the leaf tip. The redox state of the primary electron acceptor, Q, of photosystem II was estimated using a non-invasive method. Distal mature leaf sections displayed typical F680 induction curves which were generally anti-parallel with CO₂ fixation and during which Q became gradually oxidised. In leaf-base sections net assimilation of CO₂ was not detectable, F680 quenched slowly and monotonously without displaying any of the oscillations typical of mature tissue and Q remained relatively reduced. Sections cut from mid-regions of the leaf showed intermediate characteristics. There were no major differences between the wheat and maize leaf in the parameters measured. The results support the hypothesis that generation of the transthylakoid proton gradient and associated ATP production is not a major limitation to photosynthesis during leaf development in either C₃ or C₄ plants. Removal of CO₂ from the mature leaf sections caused little change in steady-state F680 and produced about 50% reduction of Q. When O₂ was then removed, F680 rose sharply and Q became almost totally reduced. In immature tissue unable to assimilate CO₂, removal of O₂ alone caused a similar large rise in F680 and reduction of Q whilst removal of CO₂ had negligible effects on F680 and the redox state of Q. It is concluded that in leaf tissue unable to assimilate CO₂, either because CO₂ is absent or the tissue is immature, O₂ acts as an electron acceptor and maintains Q in a partially oxidised state. The important implication that O₂ may have a role in the prevention of photoinhibition of the photochemical apparatus in the developing leaf is discussed.Abbreviations F680 chlorophyll fluorescence emission at 680 nm - PSI photosystem I - PSII photosystem II - Q PSII primary electron acceptor - pH transthylakoid proton gradient     

Non-invasive monitoring of photosynthetic activity and water content in forest lichens by spectral reflectance data and RGB colors from photographs

There is a need for non-invasive monitoring of temporal and spatial variation in hydration and photosynthetic activity of red-listed poikilohydric autotrophs. Here, we simultaneously recorded kinetics in RGB-colors (photos), reflectance spectra, water content, maximal (F_V/F_M), and effective quantum yield of PSII (Φ_PSII) during desiccation in foliose lichens differing in cortical characteristics and photobionts. The spectral absorbance peaks of chlorophyll a, phycocyanin, and phycoerythrin were clearly displayed at high hydration levels. Brightness and total RGB colors of the lichens strongly increased during desiccation. The normalized difference vegetation index (NDVI) efficiently estimated hydration level and Φ_PSII – a proxy for lichen photosynthesis – in all species, including threatened old forest lichens. Color and reflectance indices based on green wavelengths gave good estimates of water content in cephalo- and chlorolichens, but not in cyanolichens with a wider range of photosynthetic pigments. Due to species-specific characteristics, species-wise calibration is essential for non-invasive assessments of lichen functioning.     

Changes in activities of antioxidant enzymes and their gene expression during leaf development of sweetpotato

To understand the functions of antioxidant enzymes during leaf development in sweetpotato, we investigated the activities of several antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POX), ascorbate peroxidase (APX) and catalase (CAT). Significant increases were observed in the activities of SOD, POX and APX during the late stage of leaf development, whereas CAT activity increased during the early developmental stage. By RT-PCR analysis, various POX and APX genes showed differential expression patterns during leaf development. Four POX genes swpa3, swpa4, swpa6, swpb4 and one APX gene swAPX1 exhibited high levels of gene expression during the senescence stage of leaf development, but two POX genes, swpa1 and swpa7 were preferentially expressed at both the mature green and the late senescence stages of leaf development. These results indicate that hydrogen peroxide (H₂O₂)-related antioxidant enzymes are differentially regulated in the process of leaf development of sweetpotato.     

Progress in understanding the origin and functions of carotenoid hydroxylases in plants

The dihydroxy xanthophylls lutein (beta,epsilon-carotene-3,3'-diol) and zeaxanthin (beta,beta-carotene-3,3'-diol) are well known for their critical roles in photosystem structure and adaptation of plants to high light stress. As a group, carotenoid hydroxylases catalyze the formation of lutein and zeaxanthin from their corresponding cyclic carotene precursors. Carotenoid beta-ring hydroxylases (beta-hydroxylases) have been isolated and characterized from various organisms and have invariably been shown to be non-heme di-iron monooxygenases. The presence of an epsilon-ring specific hydroxylase (epsilon-hydroxylase) was genetically demonstrated by isolation of mutants at the LUT1 locus in Arabidopsis. This review focuses on progress in understanding the molecular and biochemical nature of the epsilon-hydroxylase and the in vivo overlapping functions of the various carotenoid beta- and epsilon-hydroxylases in Arabidopsis.     

Spatial patterns of photosynthetic characteristics and leaf physical traits of plants in the Loess Plateau of China

The spatial patterns of photosynthetic characteristics and leaf physical traits of 171 plants belonging to nine life-forms or functional groups (trees, shrubs, herbs, evergreen trees, deciduous trees, C₃ and C₄ herbaceous plants, leguminous and non-leguminous species) and their relationships with environmental factors in seven sites, Yangling, Yongshou, Tongchuan, Fuxian, Ansai, Mizhi and Shenmu, ranging from south to north in the Loess Plateau of China were studied. The results showed that the leaf light-saturated photosynthetic rate (P_max), photosynthetic nitrogen use efficiency (PNUE), chlorophyll content (Chl), and leaf mass per area (LMA) of all the plants in the Loess Plateau varied significantly among three life-form groups, i.e., trees, shrubs and herbs, and two groups, i.e., evergreen trees and deciduous trees, but leaf nitrogen content differed little among different life-form groups. For the 171 plants in the Loess Plateau, leaf P_max was positively correlated with PNUE. The leaf nitrogen content per unit area (N_area) was positively correlated but Chl was negatively correlated with the LMA. When controlling the LMA, the N_area was positively correlated with the Chl (partial r = 0.20, P < 0.05). With regard to relationships between photosynthetic characteristics and leaf physical traits, the P_max was positively correlated with N _area, while the PNUE was positively correlated with the Chl and negatively correlated with the N_area and LMA. For all the species in the Loess Plateau, the PNUE was negatively correlated with the latitude and annual solar radiation (ASR), but positively correlated with the mean annual rainfall (MAR) and mean annual temperature (MAT). With regard to the leaf physical traits, the leaf Chl was negatively correlated with the latitude and ASR, but positively correlated with the MAR and MAT. However, the N_area and LMA were positively correlated with the latitude and ASR, but negatively correlated with the MAR and MAT. In general, leaf N_area and LMA increased, while PNUE and Chl decreased with increases in the latitude and ASR and decreases in MAR and MAT. Electronic supplementary material The online version of this article () contains supplementary material, which is available to authorized users.     

Polygalacturonase expression during leaf abscission of normal and transgenic tomato plants

Polygalacturonase (PG, EC 3.2.1.15), an enzyme commonly found in ripening fruit, has also been shown to be associated with abscission. A zone-specific rise in PG activity accompanies the abscission of both leaves and flowers of tomato (Lycopersicon esculentum Mill.) plants. Studies of transgenic plants expressing an antisense RNA for fruit PG indicate that although the enzyme activity in transgenic fruit is < 1 % of that in untransformed fruit, the PG activity in the leaf abscission zone increases during separation to a similar value to that in untransformed plants. The timing and rate of leaf abscission in transgenic plants are unaffected by the introduction of the antisense gene. A polyclonal antibody raised against tomato fruit PG does not recognise the leaf abscission protein. Furthermore a complementary DNA (cDNA) clone (pTOM6), which has been demonstrated to code for fruit PG, does not hybridise to mRNA isolated from the abscission-zone region of tomato leaves. These results indicate that the PG protein in abscission zones of tomato is different from that in the fruit, and that the gene coding for this protein may also be different.Abbreviation PG polygalacturonase The authors of this paper are grateful to David Jackson of the John Innes Institute, Norwich, UK for his assistance with the in-situ hybridisation work. This research was supported by an Agricultural and Food Research Council Post-Doctoral award to J.E.T., and by a grant to D.G. from the Science and Engineering Research Council Biotechnology Directorate in association with ICI seeds. The work was carried out under Ministry of Agriculture, Food and Fisheries licences.     

Competition between isoprene emission and pigment synthesis during leaf development in aspen

In growing leaves, lack of isoprene synthase (IspS) is considered responsible for delayed isoprene emission, but competition for dimethylallyl diphosphate (DMADP), the substrate for both isoprene synthesis and prenyltransferase reactions in photosynthetic pigment and phytohormone synthesis, can also play a role. We used a kinetic approach based on post‐illumination isoprene decay and modelling DMADP consumption to estimate in vivo kinetic characteristics of IspS and prenyltransferase reactions, and to determine the share of DMADP use by different processes through leaf development in Populus tremula. Pigment synthesis rate was also estimated from pigment accumulation data and distribution of DMADP use from isoprene emission changes due to alendronate, a selective inhibitor of prenyltransferases. Development of photosynthetic activity and pigment synthesis occurred with the greatest rate in 1‐ to 5‐day‐old leaves when isoprene emission was absent. Isoprene emission commenced on days 5 and 6 and increased simultaneously with slowing down of pigment synthesis. In vivo Michaelis–Menten constant (K_m) values obtained were 265 nmol m^?2 (20 μm ) for DMADP‐consuming prenyltransferase reactions and 2560 nmol m^?2 (190 μm ) for IspS. Thus, despite decelerating pigment synthesis reactions in maturing leaves, isoprene emission in young leaves was limited by both IspS activity and competition for DMADP by prenyltransferase reactions.     

Transfer and trapping of excitation energy in Photosystem II as studied by chlorophyll a2 fluorescence quenching by dinitrobenzene and carotenoid triplet. The matrix model

1. The curves representing the reciprocal fluorescence yield of chlorophyll a of Photosystem II (PS II) in Chlorella vulgaris as a function of the concentration of m-dinitrobenzene in the states P Q and P Q^-, are found to be straight parallel lines; P is the primary donor and Q the primary acceptor of PS II. In the weakly trapping state P Q^- the half-quenching of dinitrobenzene is about 0.2 mM, in vitro it is of the order of 10 mM. The fluorescence yield as a function of the concentration of a quencher is described for three models for the structure of pigment systems: the model of separate units, the model of limited energy transfer between the units, and the matrix model. If it is assumed that the rate constant of quenching by dinitrobenzene is high and thus the number of dinitrobenzene molecules per reaction center low, it can be concluded that the pigment system of PS II in C. vulgaris is a matrix of chlorophyll molecules in which the reaction centers are embedded. Theoretical and experimental evidence is consistent with such an assumption.

For Cyanidium caldarium the zero fluorescence yield Ф₀ and its quenching by dinitrobenzene were found to be much smaller than the corresponding quantities for C. vulgaris. Nevertheless, our measurements on C. caldarium could be interpreted by the assumption that the essential properties (rate constants, dinitrobenzene quenching) of PS II are the same for these two species belonging to such widely different groups.

2. The measured dinitrobenzene concentrations required for half-quenching in vivo and other observations are explained by (non-rate-limiting) energy transfer between the chlorophyll a molecules of PS II and by the assumptions that dinitrobenzene is approximately distributed at random in the membrane and does not diffuse during excitation.

3. The fluorescence kinetics of C. vulgaris during a 350 ns laser flash of variable intensity could be simulated on a computer using the matrix model. From the observed fluorescence quenching by the carotenoid triplet (C^T) and the measurement of the number of C^T per reaction center via difference absorption spectroscopy, the rate constant for quenching of C^T is calculated to be k_T = 3.3 · 10¹¹ s⁻¹ which is almost equal to the rate constant of trapping by an open reaction center (Duysens, L.N.M. (1979) CIBA Foundation Symposium 61 (New Series), pp. 323–340).

4. The fluorescence quenching by C^T in non-treated spinach chloroplasts after a 500 ns laser flash (Breton, J., Geacintov, N.E. and Swenberg, C.E. (1979) Biochim. Biophys. Acta 548, 616–635) could be explained within the framework of the matrix model when the value for k_T is used as given in point 3.

5. The observations mentioned under point 1 indicate that the fluorescence yield Ф₀ for centers in trapping state P Q is probably for a fraction exceeding 0.8 emitted by PS II.     

Plastid structure and chlorophyll content in leaf primordia of onion bulbs

Angiosperms do not synthesize chlorophyll in the dark. Here we show that leaf primordia in onion bulbs are green, though they developing in dark conditions. We present results that show plastids in green primordia are chloroplasts, and that they contain chlorophyll as well as embryos in seeds of certain angiosperms.     

Changes in activities of antioxidative system and monoterpene and photochemical efficiency during seasonal leaf senescence in Hevea brasiliensis trees

A variety of ecophysiological parameters were monitored in leaves of Hevea brasiliensis (rubber tree) during seasonal leaf senescence. Higher levels of hydrogen peroxide and malondialdehyde, and lower content of total protein and efficiency of photochemistry of photosystem II (PSII) were observed in the senescent leaves (SL) compared to the mature leaves (ML). A significant decrease in the contents of chlorophyll (Chl) and carotenoids (Car) in SL was also observed, but with increase in ratio of Car/Chl. Moreover, activities of superoxide dismutases, catalase, and glutathione reductase in SL were strongly suppressed. In contrast, the activities of guaiacol peroxidase (POD) and ascorbate peroxidase (APX), and the contents of reduced ascorbate, total ascorbate, reduced glutathione and total glutathione were considerably increased in SL compared to ML. In addition, α-pinene, β-pinene, sabinene and total monoterpene pool in SL were drastically decreased. Taken together, these results indicate that the enhanced activities of POD and APX, and further activation of ascorbate-glutathione cycle conferred an important photoprotection against oxidative stress in senescent leaves of rubber trees. The increased Car/Chl could give the protection against photoxidation as well.     

Chlorophyll fluorescence imaging of photosynthetic activity and pigment contents of the resurrection plants Ramonda serbica and Ramonda nathaliae during dehydration and rehydration

The desiccation-tolerant plants of the R. serbica and R. nathaliae are resurrection plants which are able to fully recover their physiological function after anabiosis. A comparison of chlorophyll fluorescence imaging and photosynthetic pigment contents responses of R. serbica and, for the first time, R. nathaliae to dehydration and rehydration were investigated. For this purpose, plants after collection from their natural habitats were kept fully watered for 14 days at natural condition. The experiment was conducted with mature leaves of both species. R. serbica and R. nathaliae plants were dehydrated to 5.88 % and 7.87 % relative water content (RWC) by withholding water for 15 days, afterwards the plants were rehydrated for 72 hours to 94.67 % and 97.02 % RWC. During desiccation, R. serbica plants preserved the chlorophyll content about 84 %, while R. nathaliae about 90 %. During dehydration when RWC were more than 40 %, photochemical efficiency of PSII for photochemistry, the Fv/Fm ratio, decreased about 40 % in R. nathaliae plants, but a strong reduction with 60 % was recorded for R. serbica. Following rehydration, the Fv/Fm ratio recovered more rapidly in R. nathaliae. The higher photosynthetic rates could also be detected via imaging the chlorophyll fluorescence decrease ratio Rfd, which possessed higher values after rehydration leaves of R. nathaliae as compared to R. serbica. The results showed that the photosynthetic activity and chlorophyll contents after rehydration are recovered more rapidly in R. nathaliae in comparison to R. serbica.     

Seasonal changes in photosystem II organisation and pigment composition in Pinus sylvestris

Conifers of the boreal zone encounter considerable combined stress of low temperature and high light during winter, when photosynthetic consumption of excitation energy is blocked. In the evergreen Pinus sylvestris L. these stresses coincided with major seasonal changes in photosystem II (PSII) organisation and pigment composition. The earliest changes occurred in September, before any freezing stress, with initial losses of chlorophyll, the D1-protein of the PSII reaction centre and of PSII light-harvesting-complex (LHC II) proteins. In October there was a transient increase in F₀, resulting from detachment of the light-harvesting antennae as reaction centres lost D1. The D1-protein content eventually decreased to 90%, reaching a minimum by December, but PSII photochemical efficiency [variable fluorescence (F_v)/maximum fluorescence (F_m)] did not reach the winter minimum until mid-February. The carotenoid composition varied seasonally with a twofold increase in lutein and the carotenoids of the xanthophyll cycle during winter, while the epoxidation state of the xanthophylls decreased from 0.9 to 0.1 from October to January. The loss of chlorophyll was complete by October and during winter much of the remaining chlorophyll was reorganised in aggregates of specific polypeptide composition, which apparently efficiently quench excitation energy through non-radiative dissipation. The timing of the autumn and winter changes indicated that xanthophyll de-epoxidation correlates with winter quenching of chlorophyll fluorescence while the drop in photochemical efficiency relates more to loss of D1-protein. In April and May recovery of the photochemistry of PSII, protein synthesis, pigment rearrangements and zeaxanthin epoxidation occurred concomitantly. Indoor recovery of photosynthesis in winter-stressed branches under favourable conditions was completed within 3 d, with rapid increases in F₀, the epoxidation state of the xanthophylls and in light-harvesting polypeptides, followed by recovery of D1-protein content and F_v/F_m, all without net increase in chlorophyll. The fall and winter reorganisation allow Pinus sylvestris to maintain a large stock of chlorophyll in a quenched, photoprotected state, allowing rapid recovery of photosynthesis in spring.Abbreviations Elips early light-induced proteins - EPS epoxidation state - F₀ instantaneous fluorescence - F_m maximum fluorescence - F_v variable fluorescence - LHC II light-harvesting complex of PSII - LiDS lithium dodecyl sulfate This research was supported by the Swedish Natural Science Research Council. We wish to thank Dr. Adrian Clarke¹ (Department of Plant Physiology, University of Umeå, Sweden) for advice on electrophoresis, valuable discussion and providing antibodies. Dr. Stefan Jansson¹ and Dr. Torill Hundal (Department for Biochemistry, University of Stockholm, Sweden) provided antibodies. Jan Karlsson¹ helped with the HPLC, Dr. Marianna Krol gave advice on green gels and Dr. Vaughan Hurry (Cooperative Research Centre for Plant Sciences, Australian National University, Canberra, Australia) provided valuable discussion.