How to correctly determine the different chlorophyll fluorescence parameters and the chlorophyll fluorescence decrease ratio R<Subscript>Fd</Subscript> of leaves with the PAM fluorometer

This contribution is a practical guide to the measurement of the different chlorophyll (Chl) fluorescence parameters and gives examples of their development under high-irradiance stress. From the Chl fluorescence induction kinetics upon irradiation of dark-adapted leaves, measured with the PAM fluorometer, various Chl fluorescence parameters, ratios, and quenching coefficients can be determined, which provide information on the functionality of the photosystem 2 (PS2) and the photosynthetic apparatus. These are the parameters F_v, F_m, F₀, F_m′, F_v′, NF, and ΔF, the Chl fluorescence ratios F_v/F_m, F_v/F₀, ΔF/F_m′, as well as the photochemical (q_P) and non-photochemical quenching coefficients (q_N, q_CN, and NPQ). q_N consists of three components (q_N = q_E + q_T + q_I), the contribution of which can be determined via Chl fluorescence relaxation kinetics measured in the dark period after the induction kinetics. The above Chl fluorescence parameters and ratios, many of which are measured in the dark-adapted state of leaves, primarily provide information on the functionality of PS2. In fully developed green and dark-green leaves these Chl fluorescence parameters, measured at the upper adaxial leaf side, only reflect the Chl fluorescence of a small portion of the leaf chloroplasts of the green palisade parenchyma cells at the upper outer leaf half. Thus, PAM fluorometer measurements have to be performed at both leaf sides to obtain information on all chloroplasts of the whole leaf. Combined high irradiance (HI) and heat stress, applied at the upper leaf side, strongly reduced the quantum yield of the photochemical energy conversion at the upper leaf half to nearly zero, whereas the Chl fluorescence signals measured at the lower leaf side were not or only little affected. During this HL-stress treatment, q_N, q_CN, and NPQ increased in both leaf sides, but to a much higher extent at the lower compared to the upper leaf side. q_N was the best indicator for non-photochemical quenching even during a stronger HL-stress, whereas q_CN and NPQ decreased with progressive stress even though non-photochemical quenching still continued. It is strongly recommended to determine, in addition to the classical fluorescence parameters, via the PAM fluorometer also the Chl fluorescence decrease ratio R_Fd (F_d/F_s), which, when measured at saturation irradiance is directly correlated to the net CO₂ assimilation rate (_{P N}) of leaves. This R_Fd-ratio can be determined from the Chl fluorescence induction kinetics measured with the PAM fluorometer using continuous saturating light (cSL) during 4–5 min. As the R_Fd-values are fast measurable indicators correlating with the photosynthetic activity of whole leaves, they should always be determined via the PAM fluorometer parallel to the other Chl fluorescence coefficients and ratios.     

Chlorophyll fluorescence imaging of photosynthetic activity with the flash-lamp fluorescence imaging system

A flash-lamp chlorophyll (Chl) fluorescence imaging system (FL-FIS) is described that allows to screen and image the photosynthetic activity of several thousand leaf points (pixels) of intact leaves in a non-destructive way within a few seconds. This includes also the registration of several thousand leaf point images of the four natural fluorescence bands of plants in the blue (440 nm) and green (520 nm) regions as well as the red (near 690 nm) and far-red (near 740 nm) Chl fluorescence. The latest components of this Karlsruhe FL-FIS are presented as well as its advantage as compared to the classical single leaf point measurements where only the fluorescence information of one leaf point is sensed per each measurement. Moreover, using the conventional He-Ne-laser induced two-wavelengths Chl fluorometer LITWaF, we demonstrated that the photosynthetic activity of leaves can be determined measuring the Chl fluorescence decrease ratio, R_Fd (defined as Chl fluorescence decrease F_d from maximum to steady state fluorescence F_s:F_d/F_s), that is determined by the Chl fluorescence induction kinetics (Kautsky effect). The height of the values of the Chl fluorescence decrease ratio R_Fd is linearly correlated to the net photosynthetic CO₂ fixation rate P _N as is indicated here for sun and shade leaves of various trees that considerably differ in their P _N. Imaging the R_Fd-ratio of intact leaves permitted the detection of considerable gradients in photosynthetic capacity across the leaf area as well as the spatial heterogeneity and patchiness of photosynthetic quantum conversion within the control leaf and the stressed plants. The higher photosynthetic capacity of sun versus shade leaves was screened by Chl fluorescence imaging. Profile analysis of fluoresence signals (along a line across the leaf area) and histograms (the signal frequency distribution of the fluorescence information of all measured leaf pixels) of Chl fluorescence yield and Chl fluorescence ratios allow, with a high statistical significance, the quantification of the differences in photosynthetic activity between various areas of the leaf as well as between control leaves and water stressed leaves. The progressive uptake and transfer of the herbicide diuron via the petiole into the leaf of an intact plant and the concomitant loss of photosynthetic quantum conversion was followed with high precision by imaging the increase of the red Chl fluorescence F₆₉₀. Differences in the availability and absorption of soil nitrogen of crop plants can be documented via this flash-lamp fluorescence imaging technique by imaging the blue/red ratio image F₄₄₀/F₆₉₀, whereas differences in Chl content are detected by collecting images of the fluorescence ratio red/far-red, F₆₉₀/F₇₄₀.     

Effects of rhizobia inoculation and nitrogen fertilization on photosynthetic physiology of soybean

Plant growth, contents of photosynthetic pigments, photosynthetic gas exchange, and chlorophyll (Chl) fluorescence in soybean [Glycine max (L.) Merr. cv. Heinong37] were investigated after it was inoculated with Sinorhizobium fredii USDA191 or treated with 5 mM (NH₄)₂SO₄ (N5) and 30 mM (NH₄)₂SO₄ (N30), respectively. In the plants following N5 fertilization, not only plant biomass, leaf area, and Chl content, but also net photosynthetic rate (P _N), stomatal conductance (g _s), carboxylation efficiency (CE), maximum photochemical efficiency (F_v/F_m) of photosystem 2 (PS2), and quantum yield of PS2 (Φ_PS2) were markedly improved as compared with the control plants. There were also positive effects on plant growth and plant photosynthesis after rhizobia inoculation, but the effects were much less than those of N5 fertilization. For N30 plants there were no significant positive effects on plant growth and photosynthetic capacity. Plant biomass, P _N, and g _s were similar to those of N-limited (control) plants. Φ_PS2 and photochemical quenching (q_P) were obviously declined while content of carotenoids and non-photochemical quenching (q_N) were significantly enhanced in N30 treated plants. This indicated that excess N supply may cause some negative effects on soybean plants.     

Effects of water stress and high temperature on gas exchange and chlorophyll fluorescence in Triticum aestivum L.

Wheat plants grown in controlled growth chambers were exposed to drought stress (DS) and high temperature (HT) singly and in combination (DS+HT). The effects of these two stresses on net photosynthetic rate (P _N), stomatal conductance (g _s), intercellular CO₂ concentration (C _i), quantum efficiency of photosystem 2 (Φ_PS2), variable to maximum chlorophyll (Chl) fluorescence (F_v/F_m), photochemical (q_p) and non-photochemical (NPQ) Chl fluorescence, and yield were investigated. Grain yield was decreased by 21 % due to DS, while it was increased by 26 % due to HT. P _N, g _s, C _i, and Chl fluorescence were dramatically reduced to DS, HT, and their interaction, except NPQ which showed an increase due to HT.     

Method for resolution and quantification of components of the non-photochemical quenching (<Emphasis Type="Italic">q</Emphasis><Subscript>N</Subscript>)

A new method of the chlorophyll (Chl) a fluorescence quenching analysis is described, which allows the calculation of values of (at least) three components of the non-photochemical quenching of the variable Chl a fluorescence (q _N) using a non-linear regression of a multi-exponential function within experimental data. Formulae for coefficients of the “energy”-dependent (ΔpH-dependent) quenching (q _E), the state-transition quenching (q _T) and the photo/inhibitory quenching (q _I) of Chl a fluorescence were found on the basis of three assumptions: (i) the dark relaxation kinetics of q _N, as well as of all its components, is of an exponential nature, (ii) the superposition principle is valid for individual Chl a fluorescence quenching processes and (iii) the same reference fluorescence level (namely the maximum variable Chl a fluorescence yield in the dark-adapted state, F _V) is used to define both q _N and its components. All definitions as well as the algorithms for analytical recognition of the q _N components are theoretically clarified and experimentally tested. The described theory results in a rather simple equation allowing to compute values for all q _N components (q _E, q _T, q _I) as well as the half-times of relaxation (τ_1/2) of corresponding quenching processes. It is demonstrated that under the above assumptions it holds: q _N = q _E + q _T + q _I. The theoretically derived equations are tested, and the results obtained are discussed for non-stressed and stressed photosynthetically active samples. Semi-empirical formulae for a fast estimation of values of the q _N components from experimental data are also given.     

Diurnal Changes of Gas Exchange,Chlorophyll Fluorescence,and Stomatal Aperture of Hybrid Poplar Clones Subjected to Midday Light Stress

Diurnal changes in net photosynthetic rate (P _N), chlorophyll (Chl) fluorescence, and stomatal aperture of several hybrid poplar clones subjected to midday light stress were measured in July and August of 1996. Midday depression of P _N, photosystem 2 (PS2) efficiency, stomatal conductance (g _s), and stomatal aperture was observed in all clones, though at differing rates among them. Non-uniform stomatal closure occurred at noon and at other times, requiring a modification of intercellular CO₂ concentration (C ₁). A linear relationship was found between g _s and stomatal aperture. More than half of the photons absorbed by PS2 centre dissipated thermally when subjected to light stress at noon. There was a linear relationship between the rate of PS2 photochemical electron transport (PxPFD) and P _N. There was a consensus for two fluorescence indicators (1 – q_P/q_N and (F_m' – F)/F_m') in assessment of susceptibility of photoinhibition in the clones. According to P _N, Chl fluorescence, and stomatal aperture, we conclude that midday depression of photosynthesis can be attributed to both stomatal and non-stomatal limitations.     

Photosynthesis in Drought-Adapted Cassava

After 45 d of limited water supply, cassava (Manihot esculenta Crantz) exhibited pronounced reduction in shoot growth, high leaf fall, and decreased stomatal conductance. However, the water status of the remaining leaves was unaffected. This was combined with an amplified heliotropic response and drooping which minimises radiant energy interception at mid-day, suggesting that leaves are sensitive to high irradiance (I). In well-irrigated plants, CO₂-saturated oxygen evolution and net photosynthetic rate (P _N) in air were markedly higher (5-fold) in young (expanding) leaves than in mature leaves. Water limitation did not strongly modify CO₂-saturated oxygen evolution but it altered P _N in air for both types of leaves, although differently. The mature leaves of drought-adapted plants displayed residual rate of P _N and deteriorated photosystem 2 (PS2) photochemistry estimated from chlorophyll (Chl) a fluorescence measurements. In young leaves at moderate I, P _N was depressed by only 66 % in stressed plants. Moreover, the photochemical quenching of Chl a fluorescence and the quantum efficiency of PS2 photochemistry in young leaves were comparable in both control and stressed plants. In contrast at high I, P _N was almost null and marked decreases in the two fluorescence parameters were apparent. Hence the strong heliotropic response and drooping displayed by young leaves under water limitation is an important strategy for avoiding inactivation of P _N by high I and therefore for cassava tolerance to drought.     

Photoprotective function of betacyanin in leaves of <Emphasis Type="Italic">Amaranthus cruentus</Emphasis> L. under water stress

The photoprotective function of leaf betacyanin in water-stressed Amaranthus cruentus plants was examined by comparing leaves of two strains which differ significantly in the amount of betacyanin. At 0, 1, and 2 days after the imposed water stress, leaves were subjected to high-light (HL) treatment to assess their photosynthetic capacity and photoinhibition susceptibility. The water stress equally reduced leaf relative water content (RWC), gas-exchange rate and chlorophyll (Chl) contents in both leaves, indicating that the severity of water stress was comparable between the strains. Consequently, the extent of photoinhibition after the HL treatment increased in both strains as water stress developed; however, it was significantly greater in acyanic leaves than in betacyanic leaves, suggesting lower photoinhibition susceptibility in the betacyanic strain. The betacyanic leaves also exhibited approximately 30% higher values for photochemical quenching coefficient (q_P) during the period of water stress despite the nonphotochemical quenching coefficient (q_N) did not differ significantly between the strains. These results may be partially explained by the increased amount of leaf betacyanin under water stress. Moreover, a decrease in Chl content in betacyanic leaves might have enhanced light screening effect of betacyanin by increasing relative abundance of betacyanin to Chl molecule. In addition, reduced Chl content increased light penetrability of leaves. As a result, the extent of photoinhibition at the deeper tissue was exacerbated and the Chl fluorescence emitted from these tissues was more readily detected, facilitating assessment of photoinhibition at deeper tissues where the effect of betacyanic light screening is considered to be most apparent. Our results demonstrated that leaf betacyanin contributes to total photoprotective capacity of A. cruentus leaves by lowering excitation pressure on photosystem II (PSII) via attenuation of potentially harmful excess incident light under water stress.     

Photosynthetic activity of poikilochlorophyllous desiccation tolerant plant <Emphasis Type="Italic">Reaumuria soongorica</Emphasis> during dehydration and re-hydration

Diurnal patterns of gas exchange and chlorophyll (Chl) fluorescence parameters of photosystem 2 (PS2) as well as Chl content were analyzed in Reaumuria soongorica (Pall.) Maxim., a perennial semi-shrub during dehydration and rehydration. The net photosynthetic rate (P _N), maximum photochemical efficiency of PS2 (variable to maximum fluorescence ratio, F_v/F_m), quantum efficiency of non-cyclic electron transport of PS2, and Chl content decreased, but non-photochemical quenching of fluorescence and carotenoid content increased in stems with the increasing of drought stress. 6 d after re-hydration, new leaves budded from stems. In the re-watered plants, the chloroplast function was restored and Chl a fluorescence returned to a similar level as in the control plants. This improved hydraulic adjustment in plant triggered a positive effect on ion flow in the tissues and increased shoot electrical admittance. Thus R. soongorica plants are able to sustain drought stress through leaf abscission and keep part of Chl content in stems.     

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 responses of radish (<Emphasis Type="Italic">Raphanus sativus</Emphasis> var. <Emphasis Type="Italic">longipinnatus</Emphasis>) plants to infection by turnip mosaic virus

Plant growth, chlorophyll (Chl) content, photosynthetic gas exchange, ribulose-1,5-bisphosphate carboxylase (RuBPCO) enzyme activity, and Chl fluorescence in radish (Raphanus sativus var. longipinnatus) plants were examined after turnip mosaic virus (TuMV) infection. Plant fresh mass, dry mass, Chl content, net photosynthetic rate (P _N), transpiration rate (E), stomatal conductance (g _s), and RuBPCO activity were significantly lower in infected plants after 5 weeks of virus infection as compared to healthy plants. The 5-week virus infection did not induce significant differences in intercellular CO₂ concentration (C _i, photochemical efficiency of photosystem 2, PS2 (F_v/F_m), excitation capture efficiency of open PS2 reaction centres (F_v'/F_m'), effective quantum efficiency of photosystem 2 (ΔF/F_m'), and photochemical quenching (q_P), but non-photochemical quenching (q_N) and alternative electron sink (AES) were significantly enhanced. Thus the decreased plant biomass of TuMV-infected plants might be associated with the decreased photosynthetic activity mainly due to reduced RuBPCO activity.     

Influence of Potassium Deficiency on Photosynthesis,Chlorophyll Content,and Chloroplast Ultrastructure of Cotton Plants

In cotton (Gossypium hirsutum L.) grown in controlled-environment growth chamber the effects of K deficiency during floral bud development on leaf photosynthesis, contents of chlorophyll (Chl) and nonstructural saccharides, leaf anatomy, chloroplast ultrastructure, and plant dry matter accumulation were studied. After cotton plants received 35-d K-free nutrient solution at the early square stage, net photosynthetic rate (P _N) of the uppermost fully expanded main-stem leaves was only 23 % of the control plants receiving a full K supply. Decreased leaf P _N of K-deficient cotton was mainly associated with dramatically low Chl content, poor chloroplast ultrastructure, and restricted saccharide translocation, rather than limited stomata conductance in K-deficient leaves. Accumulation of sucrose in leaves of K-deficient plants might be associated with reduced entry of sucrose into the transport pool or decreased phloem loading. K deficiency during squaring also dramatically reduced leaf area and dry matter accumulation, and affected assimilate partitioning among plant tissues.     

The Photosynthesis and Chlorophyll a Fluorescence in Seedlings of Kandelia Candel (L.) Druce Grown under Different Nitrogen and NaCl Controls

Kandelia candel (L.) Druce is the dominant mangrove species on the west coast of northern Taiwan. We have measured the net photosynthetic rate (P _N) and chlorophyll (Chl) a fluorescence of seedlings grown at combinations of two nitrogen (0.01 and 0.1 mM) and two NaCl (250 and 430 mM NaCl) controls. With the same nitrogen level, seedlings grown at higher salinity (HS) had a significantly lower P _N and stomatal conductance (g _s) than those at lower salinity (LS). An increase in nitrogen availability significantly elevated P _N and g _s of the LS-grown seedlings. Compared to dark adapted leaves, the maximum quantum yield of photosystem 2 (PS2) (F_v/F_m) of leaves exposed to PFDs of 1200 and 1600 μmol m^-2 s^-1 for 2 h was significantly reduced. The degree of F_v/F_m reduction differed among leaves of the four types of treated plants. Chl fluorescence quenching analysis revealed differences among the examined plants in coefficients of non-photochemical and photochemical quenching. This revised version was published online in June 2006 with corrections to the Cover Date.     

Rehydration of Sugar Beet Plants after Water Stress: Effect of Cytokinins

The possibility to improve the recovery of sugar beet plants after water stress by application of synthetic cytokinins N⁶-benzyladenine (BA) or N⁶-(m-hydroxybenzyl)adenosine (HBA) was tested. Relative water content (RWC), net photosynthetic rate (P_N), transpiration rate (E), stomatal conductance (g_s), chlorophyll (Chl) a and Chl b contents, and photosystem 2 efficiency characterized by variable to maximal fluorescence ratio (F_v/F_m) were measured in control plants, in water-stressed plants, and after rehydration (4, 8, 24, and 48 h). Water stress markedly decreased parameters of gas exchange, but they started to recover soon after irrigation. Application of BA or HBA to the substrate or sprayed on leaves only slightly stimulated recovery of P_N, E, and g_s in rehydrated plants, especially during the first phases of recovery. Chl contents decreased only under severe water stress and F_v/F_m ratio was not significantly affected by water stress applied. Positive effects of BA or HBA application on Chl content and F_v/F_m ratio were mostly not observed.     

Diurnal variation of gas exchange, chlorophyll fluorescence, and xanthophyll cycle components of maize hybrids released in different years

Diurnal variation of gas exchange, chlorophyll (Chl) fluorescence, and xanthophyll cycle components of three maize (Zea mays L.) hybrids released in different years, i.e. Baimaya (1950s), Zhongdan2 (1970s), and Nongda108 (1990s), were compared. On cloudless days, the newer hybrids always had higher net photosynthetic rate (P _N), especially at noon, than the older ones. At noon, all the hybrids decreased their maximal yield of photosystem 2 (PS2) photochemistry (F_v/F_m) and actual quantum yield of PS2 (Φ_PS2), the newer ones always showing higher values. Generally, the newer hybrids displayed higher photochemical quenching of Chl (q_P) and lower non-photochemical quenching (NPQ). The interhybrid differences in P _N may be owing to their differential photochemical efficiency. A midday depression in P _N occurred in all hybrids, which might be caused by serious photoinhibition or by decreased stomatal conductance. However, midday depression in P _N was more obvious in the older hybrids, especially when leaves were senescent. The higher de-epoxidation state of the xanthophylls was noted in older hybrids, which was confirmed by their larger NPQ. The newer maize hybrids did not need a strong de-epoxidation state since they had a better photosynthetic quantum conversion rate and a lower NPQ.     

Plant shading increases lipid peroxidation and intensifies senescence-induced changes in photosynthesis and activities of ascorbate peroxidase and glutathione reductase in wheat

Plants of spring wheat (Triticum aestivum L. cv. Saxana) were grown during the autumn. Over the growth phase of three leaves (37 d after sowing), some of the plants were shaded and the plants were grown at 100 (control without shading), 70, and 40 % photosynthetically active radiation. Over 12 d, chlorophyll (Chl) and total protein (TP) contents, rate of CO₂ assimilation (P _N), maximal efficiency of photosystem 2 photochemistry (F_V/F_P), level of lipid peroxidation, and activities of antioxidative enzymes ascorbate peroxidase (APX) and glutathione reductase (GR) were followed in the 1_st, 2_nd, and 3_rd leaves (counted according to their emergence). In un-shaded plants, the Chl and TP contents, P _N, and F_V/F_P decreased during plant ageing. Further, lipid peroxidation increased, while the APX and GR activities related to the fresh mass (FM) decreased. The APX activity related to the TP content increased in the 3_rd leaves. The plant shading accelerated senescence including the increase in lipid peroxidation especially in the 1^st leaves and intensified the changes in APX and GR activities. We suggest that in the 2_nd and 3_rd leaves a degradation of APX was slowed down, which could reflect a tendency to maintain the antioxidant protection in chloroplasts of these leaves.     

A theoretical and experimental analysis of the qP and qN coefficients of chlorophyll fluorescence quenching and their relation to photochemical and nonphotochemical events

The initial (F₀), maximal (F_M) and steady-state (F_S) levels of chlorophyll fluorescence emitted by intact pea leaves exposed to various light intensities and environmental conditions, were measured with a modulated fluorescence technique and were analysed in the context of a theory for the energy fluxes within the photochemical apparatus of photosynthesis. The theoretically derived expressions of the fluorescence signals contain only three terms, X=J₂p_2F/(1–G), Y=T/(1–G) and V, where V is the relative variable fluorescence, J₂ is the light absorption flux in PS II, p_2F is the probability of fluorescence from PS II, G and T are, respectively, the probabilities for energy transfer between PS II units and for energy cycling between the reaction center and the chlorophyll pool: F₀=X, F_M=X/(1–Y) and F_S=X(1+(YV/(1–Y))). It is demonstrated that the amplitudes of the previously defined coefficients of chlorophyll fluorescence quenching, q_P and q_N, reflect, not just photochemical (q_P) or nonphotochemical (q_N) events as implied in the definitions, but both photochemical and nonphotochemical processes of PS II deactivation. The coefficient q_P is a measure of the ratio between the actual macroscopic quantum yield of photochemistry in PS II (41-1) in a given light state and its maximal value measured when all PS II traps are open (41-2) in that state, with 41-3 and 41-4. When the partial connection between PS II units is taken into consideration, 1-q_P is nonlinearily related to the fraction of closed reaction centers and is dependent on the rate constants of all (photochemical as well as nonphotochemical) exciton-consuming processes in PS II. On the other hand, 1-q_N equals the (normalized) ratio of the rate constant of photochemistry (k_2b) to the combined rate constant (k_N) of all the nonphotochemical deactivation processes excluding the rate constant k₂₂ of energy transfer between PS II units. It is demonstrated that additional (qualitative) information on the individual rate constants, k_N-k₂₂ and k_2b, is provided by the fluorescence ratios 1/F_M and (1/F₀)–(1/F_M), respectively. Although, in theory, 41-5 is determined by the value of both k_2b and k_N-k₂₂, experimental results presented in this paper show that, under various environmental conditions, 41-6 is modulated largely through changes in k _N, confirming the idea that PS II quantum efficiency is dynamically regulated in vivo by nonphotochemical energy dissipation.Abbreviations Chl chlorophyll - F₀, F_M and F_S initial, maximal and steady-state levels of modulated Chl fluorescence emitted by light-adapted leaves - PS I and II photosystem I and II - q_P and q_N (previously defined) photochemical and nonphotochemical components of Chl fluorescence quenching     

Effects of Nitrogen Deficiency on Gas Exchange,Chlorophyll Fluorescence,and Antioxidant Enzymes in Leaves of Rice Plants

Gas exchange, chlorophyll (Chl) fluorescence, and contents of photosynthetic pigments, soluble proteins (ribulose-1,5-bisphosphate carboxylase/oxygenase, RuBPCO), and antioxidant enzymes were characterized in the fully expanded 6^th leaves in rice seedlings grown on either complete (CK) or on nitrogen-deficient nutrient (N-deficiency) solutions during a 20-chase period. Compared with the control plants, the lower photosynthetic capacity at saturation irradiance (P _max) was accompanied by an increase in intercellular CO₂ concentration (C_i), indicating that in N-deficient plants the decline in P _max was not due to stomatal limitation but due to the reduced carboxylation efficiency. The fluorescence parameters _PS2, F_v/F_m, electron transport rate (ETR), and q_P showed the same tendency as P _max in N-deficient plants. Correspondingly, a higher q_N paralleled the rise of the ratio of carotenoid (Car) to Chl contents. However, F_v/F_m was still diminished, suggesting that photoinhibition did occur in the photosystem 2 (PS2) reaction centres. In addition, the activities of antioxidant enzymes on a fresh mass basis were gradually lowered, leading to the aggravation of membrane lipid peroxidation with the proceeding N-deficiency. The accumulation of malonyldialdehyde resulted in the lessening of Chl and soluble protein content. Analyses of regression showed PS2 excitation pressure (1 - q_P) was linearly correlated with the content of Chl and inversely with soluble protein (particularly RuBPCO) content. There was a lag phase in the increase of PS2 excitation pressure compared to the decrease of RuBPCO content. Therefore, the increased excitation pressure under N-deficiency is probably the result of saturation of the electron transport chain due to the limitation of the use of reductants by the Calvin cycle. Rice plants responded to N-deficiency and high irradiance by decreasing light-harvesting capacity and by increasing thermal dissipation of absorbed energy.     

Low-temperature limitations of photosynthesis in three tropical Vigna species: A chlorophyll fluorescence study

The temperature-dependence of photosynthesis and chlorophyll (Chl) fluorescence quenching components was studied between 0 and 45°C in three tropical, chilling-sensitive Vigna species and in chilling-tolerant pea. Photosynthesis of the Vigna spp. was approx. 20% more reduced by temperatures between 7 and 30°C than in pea. The latter revealed significant changes in Chl fluorescence parameters at much lower temperature than the Vigna spp. Below 15°C, the reduction state of Q_A increased quickly in pea, while in Vigna already below 30°C, an increase of reduced Q_A was obtained. The analysis of different components of non-photochemical Chl fluorescence quenching (q_N) revealed, that in pea photoinhibitory quenching (q_I) occurred below 13°C. Below ca. 7°C, a sudden breakdown of both q_P and the fast relaxing component of q_N was observed in pea.In Vigna, susceptibility of LHC II phosphorylation or limitation of electron flow by damage to PS I, the PS II reaction centre or the water-splitting system were not responsible for the chilling-sensitivity of photosynthesis between 5 and 30°C. Instead, photosynthesis was gradually limited by an inefficient use of reduction equivalents. This, in turn may increase susceptibilty to photoinhibition, which occurred below 20°C in Vigna. The combined study of q_P and of the different components of q_N allowed the demonstration of the subsequent occurrence of different limiting processes with decreasing temperature in the chilling-sensitive Vigna species.     

Acclimation of winter wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis>, cv. Yangmai 13) to low levels of solar irradiance

Winter wheat is a grass species widely planted in northern and central China, where the increase of aerosols, air pollutants and population density are causing significant reduction in solar irradiance. In order to investigate the adaptation of winter wheat (Triticum aestivum L., cv. Yangmai 13) to low irradiance conditions occurring in the downstream plain of the Yangtze River (China), plants were subjected to four solar irradiance treatments (100%, 60%, 40%, and 20% of environmental incident solar irradiance). Significant increases in chlorophyll (Chl) and xanthophyll (Xan) pigments, and decreases in Chl a/b and Xan/Chl ratios were observed in plants under low light. Light-response curves showed higher net photosynthetic rates (P _N) in fully irradiated plants, that also showed a higher light-compensation point. Shaded plants maintained high values of minimal fluorescence of dark-adapted state (F_o) and maximum quantum efficiency of PSII photochemistry (F_v/F_m) that assess a lower degree of photoinhibition under low light. Reduced irradiance caused decreases in effective quantum yield of PSII photochemistry (Φ_PSII), electron transport rate (ETR), and nonphotochemical quenching coefficient (q_N), and the promotion of excitation pressure of PSII (1 − q_P). The activities of the antioxidant enzymes superoxide dismutase and peroxidase were high under reduced light whereas no light-dependent changes in catalase activity were observed. Thiobarbituric acid reactive species content and electrolyte leakage decreased under shaded plants that showed a lower photooxidative damage. The results suggest that winter wheat cv. Yangmai 13 is able to maintain a high photosynthetic efficiency under reduced solar irradiance and acclimates well to shading tolerance. The photosynthetic and antioxidant responses of winter wheat to low light levels could be important for winter wheat cultivation and productivity.