Chlorophyll Fluorescence Parameters: The Definitions,Photosynthetic Meaning,and Mutual Relationships

Chlorophyll fluorescence parameters (Chl FPs) derived from the slow (long-term) induction kinetics of modulated Chl a fluorescence are reviewed and analysed with respect to their application in photosynthesis research. Only four mutually independent Chl FPs, calculated from values of five essential Chl fluorescence (ChlF) yields, are distinguished as the basic ones. These are: the maximum quantum yield of PS2 photochemistry (_{P O}), the photochemical quenching of variable ChlF (q_P), the non-photochemical quenching of variable ChlF (q_N), and the relative change of minimum ChlF (q_O). _{P O} refers to the dark-adapted state of a thylakoid membrane, q_P, q_N and q_O characterise the light-adapted state. It is demonstrated that all other Chl FPs can be determined using this quartet of parameters. Moreover, three FPs related to the non-radiative energy dissipation within thylakoid membranes are evaluated, namely: the non-photochemical ChlF quenching (NPQ), the complete non-photochemical quenching of ChlF (q_CN), and the effective quantum yield of non-photochemical processes in PS2 (_N). New FPs, the total quenching of variable ChlF (q_TV) and the absolute quenching of ChlF (q_A) which allow to quantify co-action of the photochemical and non-photochemical processes during a light period are defined and analysed. The interpretation of Chl FPs and recommendations for their application in the photosynthesis research are also given. Some alternative FPs used in the laboratory practice have only an approximate character and can lead to incorrect conclusions if applied to stressed plants. They are reviewed and compared with the standard ones. All formulae and conclusions discussed herein are verified using experimental values obtained on young seedlings of the Norway spruce (Picea abies [L.] Karst.).     

Photosynthetic Gas Exchange and Chlorophyll a Fluorescence of Three Wild Soybean Species in Response to NaCl Treatments

Responses of photosynthetic gas exchange and chlorophyll (Chl) a fluorescence of three wild soybeans, Glycine soja, G. tomentella, and G. tabacina occurring in different habitats of Taiwan, to four NaCl treatments, 0S, LS, MS, and HS (i.e. 0, 17, 51, and 85 mM NaCl) were compared. In G. soja following exposure to NaCl treatment for one month, the photon saturated photosynthetic rate (P _N), the ratio of variable to maximum fluorescence (F_v/F_m), the quantum yield of photosystem 2 (_PS2), and the electron transport rate (ETR) decreased dramatically. These reductions increased with increasing concentration of NaCl treatment. Plants of MS and HS treatments did not survive after extending the treatment to two months. Reductions in P _N, _PS2, and ETR (but not in F_v/F_m) were found in G. tabacina after two months of exposure to MS and HS treatments, but the reduction was not as severe as that in G. soja. In G. tomentella, significant reductions in P _N and g _s were found only in HS plants after two months of treatment, but no significant differences in F_v/F_m, _PS2, and ETR were found among plants of the four treatments. Thus the three wild soybeans in Taiwan have differentiated in their photosynthetic susceptibility to salinity, G. tomentella being the least susceptible, G. soja the most sensitive, and G. tabacina the intermediate. Different mechanisms are attributed to the inhibition effect of salinity on photosynthesis of the three species.     

Chlorophyll a fluorescence imaging of ozone-stressed Brassica napus L. plants differing in glucosinolate concentrations

Brassicaceae are characterised by glucosinolates (GS), which appear to be involved not only in biotic but also in abiotic stress responses of plants. We investigated the effect of O (3) stress on leaf GS concentrations in two lines of BRASSICA NAPUS L., differing in GS content. Ozone fumigation decreased GS concentrations in leaves of B. NAPUS of one line. In control conditions, chlorophyll content, rates of saturating photosynthesis, and quantum yield of photosystem 2 differed between the two BRASSICA lines, but differences were smaller in O (3)-stress conditions, suggesting that the relationship between leaf GS concentration and sensitivity to abiotic stress merits further research. In agreement with other ecophysiological measurements, chlorophyll fluorescence imaging clearly distinguished both lines and in some cases also treatments. A method for analysis of fluorescence images accounting for the two-dimensional leaf heterogeneity is presented.     

Chlorophyll a Fluorescence Emission,Xanthophyll Cycle Activity,and Net Photosynthetic Rate Responses to Ozone in Some Foliose and Fruticose Lichen Species

The lichens Parmelia quercina, Parmelia sulcata, Evernia prunastri, Hypogymnia physodes, and Anaptychia ciliaris were exposed to ozone (O₃) in controlled environment cuvettes designed to maintain the lichens at optimal physiological activity during exposure. Measurements of gas exchange, modulated chlorophyll (Chl) fluorescence, and pigment analysis were conducted before and after exposure to 300 mm³ (O₃) m^–3, 4 h per d for 14 d. No changes in the efficiency of photosystem 2 (PS2) photochemistry, the reduction state of Q_A, or the electron flow through PS2, measured by Chl fluorescence, were detected in any of the five lichen species studied. Additionally, neither photosynthetic CO₂ assimilation nor xanthophyll cycle activity or photosynthetic pigment concentration were affected by high O₃ concentrations. Thus the studied lichen species have significant capacities to withstand oxidative stresses induced by high concentration of O₃.     

Fluorescence Transients as Indicator for the Existence of Regulatory Mechanisms in Photosynthetic Electron Transport

In green algae several characteristic differences in the slope of the fast 685 nm fluorescence transient indicate the existence of different mechanisms for the regulation of the photosynthetic electron transport in vivo with respect to the requirements for ATP and NADPH. Autotrophically cultivated Chlamydobotrys stellata exhibits a normal time curve of the fluorescence yield. Anaerobiosis and C0₂-deficiency raise the O-, I- and S-level, whereas the P- level is lowered and the I-D-decay disappears. The readdition of oxygen increases the fluorescence significantly. Supplementation of aerobic cells with CO₂ restores the normal fluorescence transients. The replacement of carbon dioxide by acetate as a carbon source in the light lowers the overall fluorescence emission and abolishes the D-P-increase and the P-S-decline. The presence of DCMU increases fluorescence only at high intensities of incedent light. Anaerobiosis in these photoheterotrophic algae lowers the fluorescence emission. In this case DCMU increases fluorescence even at low light intensities. In Gonium multicoccum, which shows a normal fluorescence transient when cultivated autotrophically, CO₂-deficiency abolishes the O-level and increases the I- and S-niveau. Additional anaerobiosis in CO₂-deficient cells raises the steady state emission. Readdition of oxygen to these cells raises the I- and S-level even more and prevents the build up of the P-level. In Gonium     

Changes in the Non-Photochemical Quenching of Chlorophyll Fluorescence During Aging of Wheat Flag Leaves

Non-photochemical quenching of chlorophyll fluorescence (q_N) and its three components (q_Nf, q_Nm, and q_Ns) in the flag leaves of wheat grown in the field were studied by a fluorometer PAM-2000 on clear days. The diurnal variation patterns of q_N in just fully extended (JFEL) and aging leaves (AL) were similar, but q_Nm declined markedly in JFEL while it remained at a relatively high level in AL under strong sunlight at noon. Furthermore, at midday q_Nf was higher than q_Ns in JFEL, but much lower in AL. The results show the relative contributions of different mechanisms in preventing the photosynthetic apparatus from photodamage change during leaf development. This revised version was published online in June 2006 with corrections to the Cover Date.     

How Wind Affects the Photosynthetic Performance of Trees: Quantified with Chlorophyll a Fluorescence and Open-Top Chambers

Meteorological parameters inside and outside an open-top chamber (OTC) fumigation facility were recorded and the primary photosynthetic response of four tree species measured with chlorophyll (Chl) a fluorescence emission. Parameters extracted from the Chl a fluorescence transient were used to calculate photosynthetic activity of the leaves using a performance index. Measurements were made during the night throughout a single growing season. The seasonal primary photosynthetic performance in all species was significantly altered by growth in the OTCs, and the degree of response was dependent upon the species. Wind was an important effectual component of the altered environment. The average temperature was consistently 1.94±0.70 °C higher within the OTCs, whereas wind speed fluctuated substantially more between inside and outside the OTCs (0 to 8 m ⁻¹). There was a correlation between the photosynthetic performance index and wind speed in Fagus sylvatica, Fraxinus excelsior, and Prunus serotina. The response to wind was also particular to each species; the photosynthetic performance of F. sylvatica increased with wind speed (1 to 7 m s⁻¹), decreased with F. excelsior (0 to 6.5 m s⁻¹) and P. serotina (0 to 5.5 m s⁻¹). Abies alba, in contrast, was almost insensitive to wind. A model was proposed and tested for the conversion of the photosynthetic performance values collected in OTCs to predict the photosynthetic performance outside OTCs. The wide variety of responses to wind and temperature of the four species conformed to linear functions that describe the relationship of the wind speed and temperature responses with the difference in photosynthetic performance between the OTC and open environments. Specific coefficients for wind and temperature were proposed. The photosynthetic response to wind of each species depends on its ecophysiological specialisation. This revised version was published online in August 2006 with corrections to the Cover Date.     

拟南芥叶黄素缺失突变体叶绿素荧光猝灭的特性

研究了3个叶黄素组分缺失的拟南芥核基因突变体,npq₁（缺乏玉米黄质Z和单环氧玉米黄质A）、lut₂（缺乏lutein）和lut₂-npq₁（双突变体,同时缺Z和lutein）,及其对照野生型（WT）在强光诱导下叶绿素荧光猝灭的特性.与WT相比,3个突变体的叶绿素a/b没有明显的差异,F_v/F_m则有不同幅度的增加,缺乏lutein的突变体lut₂和lut₂-npq₁的叶黄素循环库（V+A+Z）显著增大.缺乏Z的突变体npq₁和lut₂-npq₁在强光下,荧光的非光化学猝灭（NPQ）的诱导受到明显抑制,lut₂的NPQ形成也受到部分抑制.强光处理9 min后,3个突变体和WT的NPQ大小顺序为WT＞lut₂＞npq₁＞npq₁-lut₂.强光诱导过程中突变体的光化学猝灭(qP)都小于WT.强光下突变体显示较弱的抗光抑制能力,其抗光抑制能力的强弱顺序为：WT＞lut₂＞npq₁＞lut₂-npq₁.结果表明叶黄素循环不但与NPQ的形成直接相关也与qP有关.     

Relationship Between Photosystem 2 Electron Transport and Photosynthetic CO2 Assimilation Responses to Irradiance in Young Apple Tree Leaves

The responses to irradiance of photosynthetic CO₂ assimilation and photosystem 2 (PS2) electron transport were simultaneously studied by gas exchange and chlorophyll (Chl) fluorescence measurement in two-year-old apple tree leaves (Malus pumila Mill. cv. Tengmu No.1/Malus hupehensis Rehd). Net photosynthetic rate (P _N) was saturated at photosynthetic photon flux density (PPFD) 600-1 100 (mol m^-2 s^-1, while the PS2 non-cyclic electron transport (P-rate) showed a maximum at PPFD 800 mol m^-2 s^-1. With PPFD increasing, either leaf potential photosynthetic CO₂ assimilation activity (F_d/F_s) and PS2 maximal photochemical activity (F_v/F_m) decreased or the ratio of the inactive PS2 reaction centres (RC) [(F_i – F_o)/(F_m – F_o)] and the slow relaxing non-photochemical Chl fluorescence quenching (q_s) increased from PPFD 1 200 mol m^-2 s^-1, but cyclic electron transport around photosystem 1 (RFp), irradiance induced PS2 RC closure [(F_s – F_o)/F_m – F_o)], and the fast and medium relaxing non-photochemical Chl fluorescence quenching (q_f and q_m) increased remarkably from PPFD 900 (mol m^-2 s^-1. Hence leaf photosynthesis of young apple leaves saturated at PPFD 800 mol m^-2 s^-1 and photoinhibition occurred above PPFD 900 mol m^-2 s^-1. During the photoinhibition at different irradiances, young apple tree leaves could dissipate excess photons mainly by energy quenching and state transition mechanisms at PPFD 900-1 100 mol m^-2 s^-1, but photosynthetic apparatus damage was unavoidable from PPFD 1 200 mol m^-2 s^-1. We propose that Chl fluorescence parameter P-rate is superior to the gas exchange parameter P _N and the Chl fluorescence parameter F_v/F_m as a definition of saturation irradiance and photoinhibition of plant leaves.     

Hydrogen Peroxide Inhibits Photosynthetic Electron Transport in Cells of Cyanobacteria

The effect of H₂O₂ on photosynthetic O₂ evolution and photosynthetic electron transfer in cells of cyanobacteria Anabaena variabilis and Anacystis nidulans was studied. The following experiments were performed: 1) directly testing the effect of exogenous H₂O₂; 2) testing the effect of intracellular H₂O₂ generated with the use of methyl viologen (MV); 3) testing the effect of inhibiting intracellular H₂O₂ decomposition by salicylic acid (SA) and 3-amino-1,2,4-triazole (AT). H₂O₂ inhibited photosynthetic O₂ evolution and light-induced reduction of p-benzoquinone (BQ) + ferricyanide (FeCy) in the Hill reaction. The I₅₀ value for H₂O₂ was 0.75 mM. Photosynthetic electron transfer in the cells treated with H₂O₂ was not maintained by H₂O₂, NH₂OH, 1,5-diphenylcarbazide, tetraphenylboron, or butylated hydroxytoluene added as artificial electron donors for Photosystem (PS) II. The H₂O CO₂, H₂O MV (involving PSII and PSI) and H₂O BQ + FeCy (chiefly dependent on PSII) electron transfer reactions were inhibited upon incubation of the cells with MV, SA, or AT. The N,N,N",N"-tetramethyl-p-phenylenediamine MV (chiefly dependent on PSI) electron transfer was inhibited by SA and AT but was resistant to MV. The results show that H₂O₂ inhibits photosynthetic electron transfer. It is unlikely that H₂O₂ could be a physiological electron donor in oxygenic photosynthesis.     

Changes in the Non-Photochemical Quenching of Chlorophyll Fluorescence During Aging of Wheat Flag Leaves

Ultrastructural changes in chloroplasts of primary leaves of 15-d-old bean plants (Phaseolus vulgaris L. cv. Cheren Starozagorski) in response to a single stress (increasing water deficit, WD) as well as to combined stress (WD plus high temperature, WD+HT) were investigated under the possible protective or reparatory effects of the carbamide cytokinin 4-PU-30 [N-(2-chloro-4-pyridyl)-N-phenylurea] applied before or after the stress. Essential structural changes in chloroplast ultrastructure occurred mainly in plants that had experienced WD+HT: the thylakoids were swollen, the envelope was destroyed, and the spatial orientation of inner membrane system was not typical. Changed starch accumulation was also observed. 4-PU-30 protected chloroplast ultrastructure under WD+HT.     

Influence of Drought,High Temperature,and Carbamide Cytokinin 4-PU-30 on Photosynthetic Activity of Bean Plants. 1. Changes in Chlorophyll Fluorescence Quenching

Fifteen-day-old bean plants (Phaseolus vulgaris L.) grown in a climatic chamber were exposed to water deficit (WD) and high temperature (HT) stresses applied separately or in combination. Changes in chlorophyll fluorescence quenching were investigated. Bean plants that endured mild (42 °C, 5 h for 2 d) WD separately or in combination with HT did not change their q_P and q_N quenching (measured at 25 °C) compared with those of the control. After 5 min testing at 45 °C, q_P in control and droughted plants strongly decreased, while q_P of plants that experienced combined WD+HT stress was insignificantly influenced, suggesting the acclimation effect of HT treatments. At more severe stresses (after 3 d-treatment), q_P measured at 25 °C was the lowest in WD+HT plants and q_N values were the highest. But when measured at 45 °C, q_P of WD+HT plants had practically the same values as at 25 °C. Under these conditions q_P of WD plants also showed an adaptation to HT. Twenty-four hours after recovery, the unfavourable effects of the stresses were strongly reduced when measured at 25 °C, but they were still present when measured at 45 °C. Positive effect of the carbamide cytokinin 4-PU-30 was well expressed only in droughted plants. This revised version was published online in June 2006 with corrections to the Cover Date.     

氮素水平对不同品种茶树光合及叶绿素荧光特性的影响

为探明氮素水平对不同品种茶树的光合系统的影响机制,以‘福鼎大白茶’、‘保靖黄金茶1号’、‘白毫早’两年生茶苗为材料,设置不施氮N_0(0g)、低氮N_1(11g)、中氮N_2(22g)和高氮N_3(33g)4个氮素[(NH_4)_2SO_4]水平的盆栽实验,研究了铵态氮对3个品种茶树的生长势、叶片叶绿素含量、光合参数与叶绿素荧光参数的影响。结果表明:(1)施氮处理能够显著促进茶树的生长,提高茶树叶片叶绿素含量、净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr),降低胞间CO_2浓度(Ci),并以N_2处理最好,但水分利用率(WUE)在3个品种茶树间表现不同。(2)在N_2处理下,3个茶树品种的叶片光系统Ⅱ(PSⅡ)暗适应下的最大光化学效率(F_v/F_m)、光化学猝灭系数(qP)、PSⅡ的相对电子传递速率(rETR)亦增加最大,非光化学淬灭系数(NPQ)降低。(3)茶树叶片叶绿素含量与光合参数间存在着一定的联系,并且具有品种特异性。研究发现,适量施氮能够显著增加茶树叶绿素含量、气孔导度、光合活性,从而使得各品种茶树净光合速率增加;氮素水平对各茶树品种的光合及荧光特性影响存在差异,水分利用率亦具有品种特异性;生产中应综合叶绿素含量、光合作用参数、叶绿素荧光参数,可快速、直观地评价不同品种茶树对氮素营养的内在需求,为茶园施肥管理提供指导。     

Estimation of Photorespiration Rate by Simultaneous Measurements of CO2, Gas Exchange Rate,and Chlorophyll Fluorescence Quenching in the C3 Plant Vigna Radiata (L.) Wilczek and the C4 Plant Amaranthus Mongostanus L.

So far the photorespiration rate (R _P) in a leaf has been determined as the difference between the net photosynthetic rates (P _N) measured in 21 % O₂ air (P _N21%) and 3 % O₂ air (P _N3%). In the C₃ plant Vigna radiata and the C₄ plant Amaranthus mongostanus L., P _N and chlorophyll fluorescence quenching in leaves were monitored simultaneously. R _P of leaves in situ was estimated as termed R _PE from the electron transport rates through photosystem 2 (PS2), and compared with R _PO (P _N3% – P _N21%). In V. radiata R _PO was 11.9 µmol(CO₂) m^–2 s^–1 and the ratio of R _PO to P _N21% was 42.2 %, whereas the ratio of R _PE to P _N21% was 25.7 %. This suggests that R _PO may be over-estimated for the real R _P in normal air. In A. mongostanus, P _N was almost not changed with a decrease in O₂ concentration from 21 to 3 %, whereas the quantum yield of PS2 was evidently affected by the change in O₂ concentration. This fact shows the presence of photorespiration in this C₄ species, where R _PE was equivalent to 3.8 % of P _N21%.     

Greening of Peas: Parallel Measurements of 77 K Emission Spectra,OJIP Chlorophyll a Fluorescence Transient,Period Four Oscillation of the Initial Fluorescence Level,Delayed Light Emission,and P700

Irradiation of etiolated leaves leads to their greening. Although this problem has a long history, the question of whether the intermittent irradiation (IMI) grown plants have fully functional reaction centres as well as the oxygen clock, before exposure to continuous irradiation (CI), had not been resolved. To answer this question, as well as to analyze the development of the two photosystems, the following parallel measurements were made: (1) Emission spectra at 77 K; (2) OJIP chlorophyll (Chl) a fluorescence transient; (3) period 4 oscillation in the flash number dependence of initial fluorescence F₀ (at 50 μs) and F_J (at 2 ms); and (4) P700. In the 1-ms-flash (FL) grown pea, that has a different biogenesis of the photosynthetic apparatus, delayed light emission (DLE) and Chl a fluorescence transient were measured in parallel. Quantitative analysis of Chl a fluorescence values provided the following conclusions: (1) IMI, not FL, plants have almost fully developed reaction centres and the oxygen clock. (2) Further greening of IMI plants under CI involves two phases: (a) during 3-4 h of CI, the number of PS2 units and connectivity between them increase, and then (b) light-harvesting antenna increases. (3) In FL, 10 min CI activates fully the oxygen clock. This revised version was published online in June 2006 with corrections to the Cover Date.     

生长调节剂对黄连木光合生理指标和荧光参数的影响

以黄连木幼苗为试材,采用叶片喷施的方法,研究了生长调节剂多效唑和矮壮素对黄连木叶片解剖结构、色素含量、光合生理指标和叶绿素荧光参数的影响,以揭示生长调节剂对黄连木光合特性的影响及其内在机制。结果显示:随着两种生长调节剂处理浓度的增加,黄连木幼苗叶片厚度、栅栏组织厚度、海绵组织厚度、色素含量、净光合速率(Pn)、PSⅡ原初光能转化效率(Fv/Fm)、PSⅡ的潜在活性(Fv/F0)、叶片单位面积吸收的能量(ABS/CS)和捕获的能量(TRo/CS)以及电子传递的能量(ETo/CS)均呈先增大后减小的趋势,并均在800mg/L多效唑和500mg/L矮壮素处理时达到最大值,且显著提高了黄连木叶片的净光合速率。研究发现,黄连木幼苗叶片栅/海比例增大和叶绿素含量的增加使叶片对光能的捕获和吸收能力提高,而PSⅡ反应中心开放比例的增大与其活性的增强保证了叶片对光能的高效传递,同时降低了能量的热耗散,是提高苗木光合速率和保证苗木光合作用高效进行的内在机制。     

Effects of arsenic on phosphorus content in different organs and chlorophyll fluorescence in primary leaves of soybean

The effect of arsenic (32 – 96 μM) on the phosphorus content and Chl fluorescence was studied in soybean (Glycine max Merril) grown in the nutrient solution with and without phosphorus. The increased concentration of As led to the decrease in P content in plant organs. Parameters of Chl fluorescence of soybean leaves in the presence of these As concentrations did not show significant changes.     

Heat Stress and Spermidine: Effect on Chlorophyll Fluorescence in Tomato Plants

Two tomato (Lycopersicon esculentum L.) cultivars: Robin (tolerant) and Roma (sensitive to heat stress) were studied. Chlorophyll fluorescence induction parameters (F_v/F_p, A_max, and Rfd) at 25 °C showed that the PS2 activity was similar for both cultivars. The parameters, measured at 38 °C, decreased in both cultivars, but more in cv. Roma. Exogenous application of 4 mM spermidine improved the plant heat-resistance in both cultivars, and especially in cv. Roma. Analysis of chlorophyll fluorescence changes during linear increase in temperature showed that cv. Robin plants have higher ability to hardening and higher resistance to thermal damage of the pigment-protein complexes structure and the activity of PS2 than cv. Roma.     

Chlorophyll Fluorescence and Photosynthetic Gas Exchange Responses to Irradiance of Tree of Heaven (Ailanthus Altissima) in Contrasting Urban Environments

Sun-and shade-adapted plants of Ailanthus altissima utilized thermal-dissipative photoprotection (NPQ) across a range of photosynthetic photon flux densities (PPFD), with higher NPQ and lower maximum quantum yield of photosystem 2 photochemistry (F_v/F_m) in sun-adapted individuals, suggesting increased engagement of antennae-based quenching. Photosynthetic quantum requirements (Q_req; number of photons per CO₂) were similar in sun and shade plants, but were low and comparable to forest understory species. Diurnal measurements showed that PPFDs in both habitats were consistently above photosynthetic compensation irradiance, and frequently exceeded saturating values. In addition, sun- and shade-adapted individuals possessed stomata that tracked short-term fluctuations in PPFD. Thus A. altissima may be unique in that it couples high, shade-plant like photosynthetic efficiency with high photosynthetic capacity in high-irradiance, while stomatal attributes that optimize water use efficiency are maintained in the shade. These features may contribute to success of A. altissima in establishing in disturbance-prone urban systems, and facilitate its spread into more PPFD-limited and competitive natural ecosystems.     

Measurement of Differences in Red Chlorophyll Fluorescence and Photosynthetic Activity between Sun and Shade Leaves by Fluorescence Imaging

With a flash-lamp chlorophyll (Chl) fluorescence imaging system (FL-FIS) the photosynthetic activity of several thousand image points of intact shade and sun leaves of beech were screened in a non-destructive way within a few seconds. The photosynthetic activity was determined via imaging the Chl fluorescence at maximum F_p and steady state fluorescence F_s of the induction kinetics (Kautsky effect) and by a subsequent determination of the images of the fluorescence decrease ratio R_Fd and the ratio F_p/F_s. Both fluorescence ratios are linearly correlated to the photosynthetic CO₂ fixation rates. This imaging method permitted to detect the gradients in photosynthetic capacity and the patchiness of photosynthetic quantum conversion across the leaf. Sun leaves of beech showed a higher photosynthetic capacity and differential pigment ratios (Chl a/b and Chls/carotenoids) than shade leaves. Profile analysis and histogram of the Chl fluorescence yield and the Chl fluorescence ratios allow to quantify the differences in photosynthetic activity between different leaf parts and between sun and shade leaves with a high statistical significance.