Simultaneous detection of photosynthetic energy storage and oxygen evolution in leaves by photothermal radiometry and photoacoustics

We have measured simultaneously the photothermal radiometry and the photoacoustic signals from intact leaves. We have confirmed that while the former senses that part of the modulated absorbed radiation not used in photosynthesis, but converted into heat, the latter, at low modulation frequencies, senses not only this heat but also the modulated oxygen evolution resulting from photosynthesis in the leaf. When photosynthetic activity is saturated upon additional excitation with strong non-modulated light, the photothermal radiometry signal increases (virtually all absorbed modulated light being converted into heat), while at the same time the photoacoustic signal decreases, because virtually no modulated oxygen evolution occurs any more. At higher modulation frequencies the behaviour of the photoacoustic signal closely follows that of the photothermal radiometry signal. We have used combined photothermal radiometry / photoacoustic measurements to estimate directly the yield of chemical energy storage in various plant species which applies for different times after excitation. Measurement of light saturation curves for wheat and Siberian pea bush leaves and of action spectra for the latter confirm the similarity between photothermal radiometry and high-frequency photoacoustic signals, and their difference from the low-frequency photoacoustic signal. Combined use of photothermal radiometry (or high-frequency photoacoustics) and low-frequency photoacoustics can thus provide more information than any one method alone. Experiments on intact chloroplasts and on a blue-green alga demonstrate that photothermal radiometry and photoacoustic methodologies can also be used for these tissues.     

In vivo measurement of spectroscopic and photochemical properties of intact leaves using the ‘mirage effect’

This paper describes a new, highly sensitive, method for in vivo studies of photosynthesis based on the ‘mirage effect’ in which thermal energy dissipation from intact leaves, illuminated with intensity-modulated light, is sensed through the periodic deflection of a laser beam propagating along the leaf surface. The photothermal deflection technique allows one to rapidly estimate the gross efficiency of photochemical energy storage by comparing the heat emission signal with and without an additional strong, photosynthetically saturating, non-modulated light. In pea leaves, the maximal storage efficiency at low light intensities was shown to approach 55%. The general utility of this simple photothermal method is illustrated by examining the variation of the deflection signal under different conditions. The spectral resolution of this new method is shown to be much higher than that of the photoacoustic method.     

The use of photothermal radiometry in assessing leaf photosynthesis: I. General properties and correlation of energy storage to P700 redox state

Energy storage measurements by modulated photothermal radiometry (PTR) were carried out on intact leaves to assess the value of the PTR method for photosynthesis research. In particular, correlations to the redox state of P₇₀₀ under various conditions were examined. PTR monitors modulated light conversion to heat by sensing the resulting modulated infra-red radiation emitted from the leaf. It is, therefore, a complementary method to photoacoustics for estimating energy storage and its time variation, particularly under controlled leaf atmosphere.With modulated light-1 (>690 nm) the energy storage approached zero and P₇₀₀ was maximally oxidized. When background light of shorter wavelength (<690 nm-light-2) was added, energy storage momentarily increased (a manifestation of Emerson enhancement) while P₇₀₀ was reduced. The values of both parameters varied as a function of the background light intensity, keeping a mutual linear relationship. Following the initial change, there was a slow reversal transient of P₇₀₀ oxidation with a parallel decrease in energy storage. Temporal correlation to P₇₀₀ redox state after dark adaptation was observed also for the energy storage measured in modulated light 2 when combined with background actinic light of medium intensity (about 50 W m²). Under these circumstances P₇₀₀ was almost totally oxidized initially and then gradually reduced while energy storage was initially low and then increased parallel to P₇₀₀ reduction.A comparison between the maximum energy storage in modulated light 1, enhanced by background light 2, to the energy storage with short wavelength light (where light tends to be more evenly distributed) indicates a comparable contribution to energy storage from each active photosystem. The above experiments indicate that energy storage contribution from PS I is directly related to the extent of openness of its reaction-centers.While some aspects of the data call for more experimentation, these experiments already establish PTR as a valuable method to monitor photosynthetic energy storage activity in vivo, particularly when used simultaneously with other non-invasive methods.Abbreviations ES energy storage - light 1 or light 2 light of spectral distribution which favors absorption in PS I or PS II, resp. - PTR photothermal radiometry - P₇₀₀ the primary donor in PS I reaction center     

A photoacoustic study of water infiltrated leaves

Photoacoustic measurements of photosynthetic energy storage were conducted on water infiltrated pea and sugar maple leaves. The samples were vacuum infiltrated with pure water or with a suitable buffer. The use of such methodology permitted an accurate determination of the energy storage parameter at low modulation frequencies, where in non-infiltrated leaves oxygen evolution dominates the photoacoustic signal and does not allow energy storage measurements. Differences between infiltration media were not essential, however the use of pure water as infiltration medium sometimes caused instability of the measured energy storage, particularly at longer experimental time. Values of energy storage in individual samples ranged mostly between 0.2 to 0.35. Measured as a function of the modulation frequency, energy storage was found to be constant from about 10 to 200 Hz for pea leaves. In sugar maple leaves, the energy storage slightly increased between 100 and 500 Hz. Obtaining an accurate value for energy storage also allowed an accurate estimation of the O₂ evolution contribution to the photoacoustic signal of an unfiltrated leaf. In a maple leaf its frequency dependence showed only the effect of diffusion in the entire frequency range (10–500 Hz). Energy storage transients were observed after long periods (ca. 1/4-2 hrs) of dark adaptation upon the transition to light. In this case the initial energy storage was roughly about 1/2 that of the steady state value indicating strong PS I activity, while PS II was transiently incompetent. Energy-storage increased during illumination in a way to correspond to photosynthetic induction events as previously measured by fluorescence and O₂ evolution. Transients in energy storage were also found following high light to low light transitions (i.e., switch off of the saturating background light), that paralleled similar transients in oxygen evolution, showing initial transient inactivation followed by progressive reactivation of PS II.Abbreviations ES energy storage - PA photoacoustic(s) - PTR photothermal radiometry     

Increased thermal deactivation of excited pigments in pea leaves subjected to photoinhibitory treatments

The photoacoustic technique was used to monitor thermal deexcitation of the photosynthetic pigments in intact pea leaves (Pisum sativum L.) submitted to photoinhibitory treatments. When the leaves were exposed to photon flux densities above 1000 micromoles per square meter per second, the amplitude of the photothermal component of the in vivo photoacoustic signal strongly increased. This high-light-induced stimulation of nonradiative energy dissipation (heat emission) was accompanied by an inverse change in the O₂ evolution activity and in the steady state emission of 685 nanometer chlorophyll fluorescence. The time course of these effects was shown to be very rapid, with a t_1/2 of around 15 minutes. When high-light-treated leaves were readapted to the dark, the heat emission changes were reversed, following somewhat slower kinetics. A reversible increase in the rate of light energy dissipation via radiationless transitions could be a photoprotective mechanism eliminating excess excitation energy from the photosynthetic reaction centers. Interestingly, this process does not operate at temperatures below about 12°C.     

Photosynthetic responses of leaves to water stress, expressed by photoacoustics and related methods : I. Probing the photoacoustic method as an indicator for water stress in vivo

The effect of leaf desiccation on the photosynthetic activities in vivo was probed by the photoacoustic method. The aim of this research was: (a) To study the photoacoustic signal per se in varied conditions in order to develop this tool as a probe for stress conditions in vivo. (b) To obtain results pertaining to electron transport activities in vivo, and confirm conclusions based on work with isolated chloroplasts, which could otherwise be the result of nonspecific damage occurring during their isolation. Leaf discs from tobacco (Nicotiana tabacum L.) were routinely used, with other species tested also for comparison. Rapid leaf desiccation caused changes in the low frequency photoacoustic signal, attributed both to the mechanism of signal transduction, influenced by changes in the structural parameters of the leaf, and to the direct (nonstomatal) inhibition of gross photosynthesis. The dependence of the photothermal part of the signal on the frequency indicated the presence of two photothermal components, one of which persisted only at low modulation frequencies (below about 100 Hz) and which largely increased with the desiccation treatment. This component was ascribed to a thermal wave which reaches the leaf surface. The other nonvariable photothermal component was ascribed to a thermal wave propagating from the chloroplasts to the surface of the mesophyll cell. Only this component is considered in the ratio of the O₂ signal to the photothermal signal, which is used to estimate the quantum yield of photosynthesis. The specific dependence of the latter ratio on the frequency yielded a comparative quantum yield parameter from its extrapolation to zero frequency, and also indicated stress induced changes in the diffusion of O₂ through the mesophyll cell, reflected by changes in its characteristic slope. The (zero frequency extrapolated) quantum yield was markedly reduced with the progression of the water stress, indicating the inhibition of (gross) phototosynthetic electron transport in vivo. This result was expressed even more emphatically by the stronger inhibition of the photochemical energy storage, obtained by photoacoustic measurements at a high modulation frequency.     

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

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

Functional aspects of the photosynthetic light reactions in heat stressed Arabidopsis deficient in digalactosyl-diacylglycerol

Plants are often submitted, in their natural environment, to various abiotic stresses such as heat stress. However, elevated temperature has a detrimental impact on overall plant growth and development. We have examined the physiological response of the dgd1-2 and dgd1-3 Arabidopsis mutants lacking 30-40% of digalactosyl-diacylglycerol (DGDG) exposed to heat constraint. These mutants, which grow similarly to wild type under normal conditions, were previously reported to be defective in basal thermotolerance as measured by cotyledon development. However their functional properties were not described. Chlorophyll fluorescence measurements and absorbance changes at 820 nm were used to monitor photosystem II (PSII) and PSI activity, respectively. It was observed that both mutants have similar photosystem activities with some differences. The mutants were less able to use near saturation light energy and elicited higher rates of cyclic PSI electron flow compare to wild type. Arabidopsis leaves exposed to short-term (5 min) mild (40 °C) or strong (44 °C) heat treatment have shown a decline in the operating effective quantum yield of PSII and in the proportion of active PSI reaction centers. However, cyclic PSI electron flow was enhanced. The establishment of the energy-dependent non-photochemical quenching of chlorophyll fluorescence was accelerated but its decline under illumination was inhibited. Furthermore, heat stress affected the process implicated in the redistribution of light excitation energy between the photosystems known as the light state transitions. All the effects of heat stress mentioned above were more intense in the mutant leaves with dgd1-3 being even more susceptible. The decreased DGDG content of the thylakoid membranes together with other lipid changes are proposed to influence the thermo-sensitivity of the light reactions of photosynthesis towards heat stress.     

The Effect of Temperature on the Photosynthesis of Ryegrass Canopies

In bright light, the rate of gross photosynthesis of ryegrasscanopies increased with increasing temperature between 10 and25 °C, in keeping with an increase in the photosynthesisof their constituent leaves. The increase in canopy gross photosynthesisoccurred irrespective of the leaf area index, which would affectthe proportion of the leaves in bright light. Because the responseof gross photosynthesis to temperature was similar to that ofrespiration, net photosynthesis also increased with temperature,irrespective of the relative magnitude of the two processes. However, these increases in photosynthesis were observed whilethe saturation deficit of water vapour was kept small (lessthan 3 g m^–3). The photosynthesis of both leaves and canopieswas reduced when measured at a greater saturation deficit; indicatingthat the increasing saturation deficit, which frequently accompaniesincreasing temperature in the field, may obscure the effectof temperature on photosynthesis. Lack of control of saturationdeficit as measurement temperature is increased may explainwhy some workers have failed to detect a positive effect oftemperature on photosynthesis. Perennial ryegrass, Loliwn perenne L., photosynthesis, temperature response, respiration, irradiance     

Correlation between chlorophyll fluorescence and photoacoustic signal transients in spinach leaves

Chlorophyll fluorescence and photoacoustic transients from dark adapted spinach leaves were measured and analyzed using the saturating pulse technique. Except for the first 30 s of photosynthetic induction, a good correlation was found between photoacoustically detected oxygen evolution at 35 Hz modulation frequency and electron flow calculated from the fluorescence quenching coefficients q_P and q_N. The induction kinetics of the photothermal signal, i.e., the photoacoustic signal at 370 Hz, reveal a fast (t _r <10 ms) and a slow (t _r 1 s) rise component. The fast component is suggested to be composed of the minimal thermal losses in photosynthesis and thermal losses from non-photosynthetic processes. The slow phase is attributed to variable thermal losses in photosynthesis. The variable thermal losses were normalized by measuring the minimal photothermal signal (H₀) in the dark-adapted state and the maximal photothermal signal (H_m) during a saturating light pulse. The kinetics of the normalized photochemical loss (H-H₀)/(H_m-H₀) obtained from high-frequency PA measurements were found to correlate with the kinetics of oxygen evolution measured at low frequency.Abbreviations F_m maximum fluorescence - F₀ initial fluorescence - F_v variable fluorescence - H photothermal signal - I in-phase - LED light emitting diode - PA photoacoustic - PL photochemical loss - Q quadrature - q_N non-photochemical quenching - q_P photochemical quenching - VCLS voltage controlled light source     

Canopy Photosynthesis and Crop Growth Rate of Eight Temperate Forage Grasses

The rates of canopy and individual leaf photosynthesis, ratesof growth of shoots and roots, and the extinction coefficientfor light of eight temperate forage grasses were determinedin the field during early autumn. Canopy gross photosynthesiswas calculated as net photosynthesis plus dark respiration adjustedfor temperature using a Q₁₀ = 2. The relationships between canopygross photosynthesis and light intensity were hyperbolic, andthe initial slopes of these curves indicated that light wasbeing utilized efficiently at low light intensities. The initialslope depended on the distribution of light in the canopy andthe quantum efficiency of the individual leaves. The maximumrate of canopy gross photosynthesis reflected the maximum rateof individual leaf photosynthesis. Although the maximum rateof canopy gross photosynthesis was correlated with crop growthrate, there was no significant relationship between daily grossphotosynthesis and crop growth rate. Indeed, daily gross photosynthesisvaried by only 22 per cent, whereas the daily growth of shootsand roots varied by 120 per cent. This poor correlation is influencedby a negative correlation (P < 0.01) between the maximumrate of canopy gross photosynthesis and the initial slope ofthe curve relating canopy gross photosynthesis and light intensity.Difficulties in the interpretation of measurements of dark respirationappeared to confound attempts to relate daily net photosynthesisto crop growth rate, individual leaf photosynthesis, and theextinction coefficient for light.     

Bill Arnold and calorimetric measurements of the quantum requirement of photosynthesis—once again ahead of his time

The approach of photocalorimetry to decide on the true quantum requirement of photosynthesis — one of the main issues of the research in the first half of the century and a source of a bitter debate — is described. Bill Arnold's original approach to get into the true answer is reflected from the point of view of present day calorimetric techniques.Abbreviations PA photoacoustic(s) - PTR photothermal radiometry - PBD probe beam deflection (thermal measurement) - ES energy storage - QR quantum requirement This is CIW/DPB Contribution No. 1286.     

Comparison of the Oxygen Exchange between Photosynthetic Cell Suspensions and Detached Leaves of Euphorbia characias L

Using a mass-spectrometric ¹⁶O₂/¹⁸O₂-isotope technique, we compared the nature and the relative importance of oxygen exchange in photomixotrophic (PM) and photoautotrophic (PA) suspensions of Euphorbia characias L. with those in intact leaves of the same species. Young and mature leaves, dividing and nondividing cell suspensions were characterized in short-term experiments. On chlorophyll basis, the gross photosynthetic activities at CO₂ saturating concentration of PA and PM suspensions varied little from those of leaves. On dry weight basis, gross photosynthesis of PA suspensions was equal to that of leaves because of their similar chlorophyll content. This was not the case in PM suspensions where gross photosynthesis was lower and largely varied during the growth cycle. The CO₂ compensation point of PA cells (155-265 parts per million) was much higher than that of leaves (50-80 ppm). Oxygen uptakes were analyzed in terms of mitochondrial respiration, photorespiration and light stimulation of oxygen uptake (LSOU), often identified to Mehlertype reactions. In PA and PM suspensions, mitochondrial respiration rates were higher than in leaves by a factor of 1.5 to 4.5. In PM suspensions, photorespiration and LSOU were observed only in nondividing cells. Photorespiration and LSOU rates were comparable in PA suspensions and leaves. Our results demonstrate that photorespiration of PA suspensions has not been affected by the 2% CO₂ concentration imposed during 2 years of culture.     

Photosynthetic energy storage in aquatic leaves measured by photothermal deflection

In a study of photosynthetic energy storage efficiency (ES), the adaxial surface of the leaves of Vallisneria americana exhibited the highest ES values (22%) of the four aquatic plants examined. V. americana leaves have a dispersed structure and it was possible to measure the energy storage properties of the epidermal cells independently of the rest of the leaf. The abaxial epidermis had a higher value of ES at zero light fluence than the adaxial epidermis but ES in the abaxial epidermis declined much more rapidly with light fluence. Thus the abaxial epidermis is more suited to lower light fluences than the adaxial epidermis. ES declined as the pH rose from 4.0 to 8.0 at a constant dissolved inorganic carbon concentration. This paralleled the change from carbon dioxide to bicarbonate and suggests that these leaves utilise CO₂ more efficiently than bicarbonate. ES increased by about 50% at pH 8.0 as leaf sections further from the leaf tip were examined which demonstrates that the older epidermal cells are less well able to use bicarbonate. Exposure to 30 min of a saturating light fluence caused the epidermal chloroplasts to move from the periclinal walls to the anticlinal walls. This decreased the photothermal signal by increasing the thermal diffusion distance and lowering the light fluence due to greater chloroplast shading. The latter effect increased ES. It appears that chloroplast movement could assist the epidermis to survive harmful light fluences.Abbreviations APW artificial pond water; atrazine- (2-chloro-4-ethylamino-6-(isopropylamine)-s-triazine) - DCMU (3-(3,4 dichlorophenyl)-1,1-dimethyl urea) - DIC dissolved inorganic carbon - DMSO dimethyl sulfoxide - ES energy storage efficiency - HEPES N-[2-Hydroxyethyl]piperazine-N-[2-ethanesulfonic acid] - MOPS 3-[N-Morpholino]propanesulfonic acid - PD photothermal deflection     

Protective and injuring action of visible light on photosynthetic apparatus in wheat plants during hyperthermia treatment

Illumination of wheat (Triticum aestivum L.) leaves during heat treatment produced either additional injury or protection of photosynthetic apparatus depending on irradiance and the heating dose. Furthermore, illumination of leaves during hyperthermia exerted differential impacts on thermal tolerances of photosynthesis and photosystem II-driven electron transport assessed from the reduction of 2,6-dichlorophenolindophenol (DCPIP). Measurements with infrared gas analyzer showed that mild heating of leaves in darkness (10 min at 38–40°C) had stronger inhibitory effect on CO₂ uptake than heating of leaves exposed to low and moderate complex irradiances (3–30 klx), as well as excessive irradiance (75–100 klx). When the leaves were heated at higher temperatures (42–44°C), the low and moderate irradiances had a protective action, while high-intensity light aggravated the inhibition of photosynthesis. Illumination of leaves with weak light during heat treatment mitigated the impairment of chloroplast ultrastructure, whereas irradiation with high-intensity light (100 klx) destroyed the sensitive population of chloroplasts. The heat-stimulated photoinhibition was stronger for leaf photosynthesis than for DCPIP reduction in chloroplasts isolated from heat-treated leaves. No correlation was observed between the extent of violaxanthin deepoxidation, zeaxanthin accumulation, and the protective effect of light on photosynthetic apparatus during heat treatments.     

Restrictions to carbon dioxide conductance and photosynthesis in spinach leaves recovering from salt stress

Salt accumulation in spinach (Spinacia oleracea L.) leaves first inhibits photosynthesis by decreasing stomatal and mesophyll conductances to CO₂ diffusion and then impairs ribulose-1,5-bisphosphate carboxylase/oxygenase (S. Delfine, A. Alvino, M. Zacchini, F. Loreto [1998] Aust J Plant Physiol 25: 395–402). We measured gas exchange and fluorescence in spinach recovering from salt accumulation. When a 21-d salt accumulation was reversed by 2 weeks of salt-free irrigation (rewatering), stomatal and mesophyll conductances and photosynthesis partially recovered. For the first time, to our knowledge, it is shown that a reduction of mesophyll conductance can be reversed and that this may influence photosynthesis. Photosynthesis and conductances did not recover when salt drainage was restricted and Na content in the leaves was greater than 3% of the dry matter. Incomplete recovery of photosynthesis in rewatered and control leaves may be attributed to an age-related reduction of conductances. Biochemical properties were not affected by the 21-d salt accumulation. However, ribulose-1,5-bisphosphate carboxylase/oxygenase activity and content were reduced by a 36- to 50-d salt accumulation. Photochemical efficiency was reduced only in 50-d salt-stressed leaves because of a decrease in the fraction of open photosystem II centers. A reduction in chlorophyll content and an increase in the chlorophyll a/b ratio were observed in 43- and 50-d salt-stressed leaves. Low chlorophyll affects light absorptance but is unlikely to change light partitioning between photosystems.     

Analysis of Light-Induced Depressions of Photosynthesis in Leaves of a Wheat Crop during the Winter

The photosynthetic performances of individual leaves of a wheat (Triticum aestivum cv Bezostaya) crop were assessed daily and throughout individual days during the winter when temperature and light levels were fluctuating. Measurements of chlorophyll fluorescence induction and the maximum quantum yield of O₂ evolution were made on individual leaves. Depressions in the ratio of variable to maximal fluorescence (F_v/F_m) were correlated with low temperatures and high light levels throughout the winter and during the course of individual days. Depressions in F_v/F_m observed in the field during the day were not accompanied by any significant change in the ability of photosystem II complexes to bind 3-(3,4-dichlorophenyl)-1-dimethyl urea, indicating that the depressions in F_v/F_m were not attributable to photodamage to the D1 protein of the photosystem II reaction center. Decreases in F_v/F_m were associated with increases in the rate of dissipation of excitation energy by radiationless decay processes and decreases in the quantum efficiency of CO₂ assimilation, indicative of a rapidly reversible light-induced “downregulation” of photosynthesis. No major changes were observed in the maximum quantum efficiency of O₂ evolution of leaves throughout periods of fluctuating temperature and light, because light-induced depressions in photosynthetic efficiency recovered within the time required to make these measurements.     

Seasonal changes in canopy photosynthesis and foliage respiration in a Rhizophora stylosa stand at the northern limit of its natural distribution

Gross photosynthesis and respiration rates of leaves at different canopy heights in a Rhizophora stylosa Griff. stand were measured monthly over 1 year at Manko Wetland, Okinawa Island, Japan, which is the northern limit of its distribution. The light-saturated net photosynthesis rate for the leaves at the top of the canopy showed a maximum value of 17 μmol CO₂ m⁻² s⁻¹ in warm season and a minimum value of 6 μmol CO₂ m⁻² s⁻¹ in cold season. The light-saturated gross photosynthesis and dark respiration rates of the leaves existing at the top of the canopy were 2−7 times and 3–16 times, respectively, those of leaves at the bottom of the canopy throughout the year. The light compensation point of leaves showed maximum and minimum peaks in warm season and cold season, respectively. The annual canopy gross photosynthesis, foliage respiration, and surplus production were estimated as 117, 49, and 68 t CO₂ ha⁻¹ year⁻¹, respectively. The energy efficiency of the annual canopy gross photosynthesis was 2.5%. The gross primary production GPP fell near the regression curve of GPP on the product of leaf area index and warmth index, the regression curve which was established for forests in the Western Pacific with humid climates.     

Computer-controlled pulse modulation system for analysis of photoacoustic signals in the time domain

A newly developed photoacoustic system for measurement of photosynthetic reactions in intact leaves is described. The system is based on pulsed light-emitting diodes, the pulse program and pulse response analysis being computer controlled. Separation of various components in the overall photoacoustic signal is achieved by curve fitting analysis of the responses following individual measuring light pulses in the millisecond time domain. This procedure is in distinction to the conventionally used analysis in the frequency domain, with the advantage that various signal components are obtained by on-line deconvolution, yielding simultaneous recordings of photothermal (complement of energy storage) and photobaric (evolution and uptake) signals. The basic components of the new system are described by block diagrams and the principal steps for deconvolution of the overall photoacoustic response are outlined. An example of application with simultaneous recording of chlorophyll fluorescence is given. It is apparent that the photobaric uptake component represents a significant part of the overall signal, particularly during induction of photosynthesis after dark-adaptation. This component probably contains not only O₂-uptake but uptake of CO₂ as well.Abbreviations PA photoacoustic - LED light-emitting-diode - RAM random access memory     

Increased photosynthetic activities and thermostability of photosystem II with leaf development of elm seedlings (Ulmus pumila) probed by the fast fluorescence rise OJIP

Experiments were conducted to investigate the photosynthetic activity and thermostability of photosystem II (PSII) in elm seedling (Ulmus pumila) leaves from initiation to full expansion. During leaf development, photosynthesis, measured as CO₂ fixation, increased gradually and reached a maximum value when leaves were fully developed. In parallel with the increase of carbon assimilation, chlorophyll content increased. The chlorophyll a fluorescence measurements showed that the maximum quantum yield of PSII primary photochemistry (φ_po), the efficiency with which the energy of trapped excitons is converted into the electron transport beyond Q_A (Ψ_o) and the quantum yield of electron transport beyond Q_A (φ_Eo) increased gradually. The low light experiments confirmed these results independently. When subjected to heat stress, young leaves exhibited progressively lower φ_po and maximal fluorescence (F_m) values with considerably higher minimal fluorescence (F_o) than mature leaves, demonstrating that PSII in newly initiating leaves is more sensitive to heat stress. Further analysis revealed that PSII structure in newly initiating leaves showed a robust alteration under heat stress, which was reflected by the clear K phase in the OJIP curves. Therefore, we suggest that the enhanced thermostability of PSII in the case of leaf growth might be associated with an improvement of the stability of the oxygen-evolving complex (OEC) to heat stress during leaf development.