An integrating sphere leaf chamber

Abstract. A combination of an optical integrating sphere and leaf chamber is described, and some principles of its design are given. It is shown that the number of quanta absorbed by the leaf inside the sphere can be found from measurements of the quantum flux density inside the sphere with and without the leaf present, when certain constant parameters of the sphere are known. Methods for finding these are given. It is not necessary to know leaf area or absorptivity for the quantum yield for photosynthesis to be derived from additional measurement of CO₂, fixation.     

Gas exchange measurements, what can they tell us about the underlying limitations to photosynthesis? Procedures and sources of error

The principles, equipment and procedures for measuring leaf and canopy gas exchange have been described previously as has chlorophyll fluorescence. Simultaneous measurement of the responses of leaf gas exchange and modulated chlorophyll fluorescence to light and CO2 concentration now provide a means to determine a wide range of key biochemical and biophysical limitations on photo synthesis in vivo. Here the mathematical frameworks and practical procedures for determining these parameters in vivo are consolidated. Leaf CO2 uptake (A) versus intercellular CO2 concentration (Ci) curves may now be routinely obtained from commercial gas exchange systems. The potential pitfalls, and means to avoid these, are examined. Calculation of in vivo maximum rates of ribulose-1,5-bisphosphate (RuBP) carboxylase/oxygenase (Rubisco) carboxylation (Vc,max), electron transport driving regeneration of RuBP (Jmax), and triose-phosphate utilization (VTPU) are explained; these three parameters are now widely assumed to represent the major limitations to light-saturated photosynthesis. Precision in determining these in intact leaves is improved by the simultaneous measurement of electron transport via modulated chlorophyll fluorescence. The A/Ci response also provides a simple practical method for quantifying the limitation that stomata impose on CO2 assimilation. Determining the rate of photorespiratory release of oxygen (Rl) has previously only been possible by isotopic methods, now, by combining gas exchange and fluorescence measurements, Rl may be determined simply and routinely in the field. The physical diffusion of CO2 from the intercellular air space to the site of Rubisco in C3 leaves has long been suspected of being a limitation on photosynthesis, but it has commonly been ignored because of the lack of a practical method for its determination. Again combining gas exchange and fluorescence provides a means to determine mesophyll conductance. This method is described and provides insights into the magnitude and basis of this limitation.     

Effects of Different CO2 Environments on the Photosynthesis-Yield Relationship and the Carbon and Water Balance of a White Clover (Trifolium repens L. cv. Blanca) Sward

Effects of atmospheric CO₂ enrichment to a level above 600 parts10^–6 on leaf and canopy gas exchange characteristics wereinvestigated in Trifolium repens, using an open system for gasexchange measurement. The cuvettes of the system served as growthchambers, allowing continuous measurement in a semi-controlledenvironment of ±350 and ±600 parts 10^–6CO₂, respectively. Carbon balance data were compared with cropyield and effects on the canopy level were compared with measuredleaf responses of photosynthesis and stomatal behaviour. Photosyntheticstimulation by high CO₂ was stronger at the canopy level (103%on average) than for leaves (90% in full light), as a consequenceof accelerated foliage area development. The latter increasedabsolute water consumption by 16%, despite strong stomatal closure.The overall result was a 63% improvement in canopy water useefficiency (WUE), while leaf WVE increased almost 3-fold insaturating light. The stomatal response was such that, whilethe internal CO₂ concentration in the leaf, ch increased withrising atmospherical CO₂ concentration, c_a, ci/c_a was somewhatdecreased. Total canopy resistance, R_c, was generally lowerat high CO₂ levels, despite higher leaf resistance. Higher canopyCO₂ loss at night and faster light extinction in a larger-sizedhigh CO₂ canopy were major drawbacks which prevented a furtherincrease in dry matter production (the harvest index was increasedby a factor 1.83). Key words: CO₂ enrichment, canopy CO₂ exchange, carbon balance, water use efficiency, leaf and canopy resistance     

Coping with branch excision when measuring leaf net photosynthetic rates in a lowland tropical forest

Measuring leaf gas exchange from canopy leaves is fundamental for our understanding of photosynthesis and for a realistic representation of carbon uptake in vegetation models. Since canopy leaves are often difficult to reach, especially in tropical forests with emergent trees up to 60 m at remote places, canopy access techniques such as canopy cranes or towers have facilitated photosynthetic measurements. These structures are expensive and therefore not very common. As an alternative, branches are often cut to enable leaf gas exchange measurements. The effect of branch excision on leaf gas exchange rates should be minimized and quantified to evaluate possible bias. We compared light-saturated leaf net photosynthetic rates measured on excised and intact branches. We selected branches positioned at three canopy positions, estimated relative to the top of the canopy: upper sunlit foliage, middle canopy foliage, and lower canopy foliage. We studied the variation of the effects of branch excision and transport among branches at these different heights in the canopy. After excision and transport, light-saturated leaf net photosynthetic rates were close to zero for most leaves due to stomatal closure. However, when the branch had acclimated to its new environmental conditions—which took on average 20 min—light-saturated leaf net photosynthetic rates did not significantly differ between the excised and intact branches. We therefore conclude that branch excision does not affect the measurement of light-saturated leaf net photosynthesis, provided that the branch is recut under water and is allowed sufficient time to acclimate to its new environmental conditions.     

An apparatus for measuring photosynthetic quantum yields and quanta absorption spectra of intact plants

Abstract An apparatus is described which consists of an assimilation chamber mounted in the centre of a light-integrating Ulbricht sphere, irradiated through two fibreoptic light pipes. This arrangement provides totally diffuse radiation in the sphere. The quantum flux density in die sphere is measured by an integrating quantaspectrometer connected to the sphere by eight fibreoptic light pipes. The quantaspectrometer gives the spectral and the total quantum flux density in the sphere. By measuring the quantum flux density in the sphere before and after the insertion of a plant the number of absorbed quanta per unit time and plant area can be determined. In addition, the spectral distribution of the absorbed quantum flux density is obtained. CO₂-assimilation per unit time and plant area is determined in the same apparatus. The totally diffuse radiation within the sphere minimizes mutual shading between branches and leaves. Hence, well-defined light-responsecurves of photosynthesis can be obtained by plotting the flux density of C0₂-assimilation as a function of the absorbed quantum flux density, and the quantum yields of intact plants can be calculated. Examples of photosynthetic quantum yield determinations and quanta absorption spectra are given for some plant species.     

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.     

The Contribution of Leaves from Different Levels within a Tomato Crop to Canopy Net Photosynthesis: An Experimental Examination of Two Canopy Models

The rates of net photosynthesis per unit ground area by a closedcanopy of tomato plants were measured over a range of naturallight flux densities. The canopy, of leaf area index 8.6, wasdivided into three horizontal layers of equal depth. On successivedays the canopy was progressively defoliated in layers fromthe ground upwards, allowing the photosynthetic contributionfrom individual leaf layers to be determined. The uppermostlayer, 23% of the total leaf area, assimilated 66% of the netCO₂ fixed by the canopy and accounted for a similar percentageof the total leaf respiration. Net photosynthesis versus light response curves for individualleaves from different positions within the canopy were alsoobtained. Leaf conductances to CO₂ transfer and the dark respirationrates of leaves from the uppermost leaf layer were approximatelyten times those from the lowest layer. The canopy data were analysed using a simple model which assumedthat the canopy was composed of leaves with identical photosyntheticand respiratory characteristics. The model fitted the data andallowed the characteristics of an ‘idealized’ leafto be estimated. The estimated values of the leaf light utilizationefficiency, ,and the leaf conductance CO₂ transfer, , were similarto values directly determined for individual leaves in the uppermostleaf layer and the estimated rate of leaf dark respiration,R_d, corresponded to measured rates for leaves much lower inthe canopy. The simple model may be used to examine gross effectsof crop environment on the leaf photosynthetic characteristicof an ‘idealized’ leaf, but cannot be used to predictaccurately canopy net photosynthesis from the photosyntheticand respiratory characteristics of any single real leaf. A moredetailed model, developed to allow explicitly for the observedvariation in and R_d within the canopy is appropriate for thispurpose.     

Photosynthesis of Ears and Flag Leaves of Wheat and Barley

Immediately after anthesis ears of spring wheat absorbed lessthan 0.5 mg CO₂, per hour in daylight and later evolved CO₂,in the light and in the dark. The rate of apparent photosynthesisof the combined flag-leaf lamina and sheath and peduncle (collectivelycalled flag leaf) of two spring wheat varieties, Atle and JufyI, was 3–4 mg per hour; the rates of the flag leaf andthe ear of two spring barleys, Plumage Archer and Proctor, wereeach about 1 mg per hour. The gas exchange of ears and flag leaves between ear emergenceand maturity accounted for most of the final grain dry weight.The CO₂, fixed by the wheat ear was equivalent to between 17and 30 per cent of the grain weight, but more than this waslost by respiration, so assimilation in the flag leaf was equivalentto 110–20 per cent of the final grain weight. In barley,photosynthesis in the flag leaf and the net CO₂ uptake by theear each provided about half of the carbohydrate in the grain. Barley ears photosynthesized more than wheat ears because oftheir greater surface, and flag leaves of wheat photosynthesizedmore than those of barley because they had more surface anda slightly greater rate of photosynthesis per dm².     

Temperature and CO2Responses of Leaf and Canopy Photosynthesis: a Clarification using the Non-rectangular Hyperbola Model of Photosynthesis

The responses of C₃leaf and canopy gross photosynthesis to increasingtemperature and CO₂can be readily understood in terms of thetemperature and CO₂dependencies of quantum yield (     

The Influence of Water Stress on the Photosynthesis of a Simulated Sward of Perennial Ryegrass

The rate of canopy photosynthesis, single leaf photosynthesis,leaf resistance to gaseous exchange, and leaf water potentialof simulated swards of perennial ryegrass (Lolium perenne cv.S24) in a controlled environment, were determined during a periodof increasing water stress and recovery from that stress. Canopyphotosynthesis did not decline immediately water was withheldbut continued at an undiminished rate for several days; thereafterit fell rapidly, particularly at first. As water stress increasedsuccessive relationships between canopy photosynthesis and irradiancebecame more curved, indicating that the effect of water stressincreased with increasing irradiance. After the swards werere-watered canopy photosynthesis rose, most rapidly during thefirst 24 h. In general, the pattern of change of leaf waterpotential was similar to that of canopy photosynthesis, althougha more detailed examination of this relationship showed it tobe hysteresial; in particular, the fall in leaf water potentialpreceded that of canopy photosynthesis. Single leaf photosynthesisappeared to be the main agent through which water stress influencedcanopy photosynthesis although in the more severely stressedswards (leaf water potentials of about—15 bars) some leaftissue died and so limited the recovery of canopy photosynthesis.The leaf resistance to gaseous diffusion increased with increasingwater stress, as did the CO₂ compensation point, thereby influencingsingle-leaf photosynthesis and through it canopy photosynthesis.     

Do epidermal lens cells facilitate the absorptance of diffuse light?

Many understory plants rely on diffuse light for photosynthesis because direct light is usually scattered by upper canopy layers before it strikes the forest floor. There is a considerable gap in the literature concerning the interaction of direct and diffuse light with leaves. Some understory plants have well-developed lens-shaped epidermal cells, which have long been thought to increase the absorption of diffuse light. To assess the role of epidermal cell shape in capturing direct vs. diffuse light, we measured leaf reflectance and transmittance with an integrating sphere system using leaves with flat (Begonia erythrophylla, Citrus reticulata, and Ficus benjamina) and lens-shaped epidermal cells (B. bowerae, Colocasia esculenta, and Impatiens velvetea). In all species examined, more light was absorbed when leaves were irradiated with direct as opposed to diffuse light. When leaves were irradiated with diffuse light, more light was transmitted and more was reflected in both leaf types, resulting in absorptance values 2-3% lower than in leaves irradiated with direct light. These data suggest that lens-shaped epidermal cells do not aid the capture of diffuse light. Palisade and mesophyll cell anatomy and leaf thickness appear to have more influence in the capture and absorption of light than does epidermal cell shape.     

Effect of Leaf Age and Shading on Photosynthesis in Potatoes (Solanum tuberosum)

The effects of leaf age and of shading on photosynthetic rateand on other leaf parameters of potato (Solanum tuberosum L.)were studied using a portable gas exchange system. A rapid decreasein the rate of photosynthesis during leaf senescence was observed.This was accompanied by an increase in stomatal resistance,and as a result a fairly constant level of sub-stomatal CO₂concentration was maintained at all leaf ages. The reductionin the photosynthetic rate in older leaves was therefore assumedto be essentially mesophyllic in origin, whereas the stomatalresponse was probably secondary. Canopy density significantly affected the rate of photosyntheticreduction with leaf age. Leaves maintained under high radiationintensities manifested a slower decline in their photosyntheticrate, especially in the early stages of their senescence, thanleaves kept under shade conditions. The latter leaves were foundto be more adapted to low radiation intensities, as indicatedby changes in their chlorophyll a:b ratio and specific leafweight Solanum tuberosum L, potato, photosynthetic rate, mesophyll, stomata, leaf age, radiation intensity, chlorophyll a:b ratio     

A Rapid Field Method for Measuring Photosynthesis with Labelled Carbon Dioxide

A portable field apparatus for the measurement of photosynthesisis described. Short pulses (20 s) of air containing ¹⁴CO₂ areapplied to circular leaf areas of 1 cm², enclosed in a transparentminiature chamber; the radioactivity of the leaf areas is measuredby liquid scintillation, using a new in-vial wet combustiontechnique. Typical daily courses of photosynthesis in maizeand cotton are presented.     

An integrated portable apparatus for the simultaneous field measurement of photosynthetic CO2 and water vapour exchange, light absorption and chlorophyll fluorescence emission of attached leaves

Abstract. A portable apparatus has been constructed to measure simultaneously the quantum yield of CO₂ assimilation, light absorption, chlorophyll fluorescence emission and water vapour exchange of attached intact leaves in the field. The core of the instrument is a light-integrating spherical leaf chamber which includes ports for a light source, photosynthetically active radiation sensor, fluorescence probes and gas inlet and outlet manifolds. Measurement of the quantum flux inside the empty chamber and with a leaf present allows determination of leaf absorptance. An open gas exchange system is employed using an infra-red analyser to measure leaf CO₂ exchange. Using a DC white light source the quantum yield of CO₂ assimilation based on absorbed light (φ_abs) may be determined rapidly in either ambient air or artificial gas mixtures. Inclusion of capacitance humidity probes into the gas inlet and outlet ports allows simultaneous determination of water vapour exchange and subsequent estimation of stomatal conductance to CO₂ and intercellular CO₂ concentration. Measurement of fluorescence emission by the sample leaf exposed to white light is achieved by a modulated fluorescence detection system. In addition to determination of the minimal, maximal and variable fluorescence levels, a further analysis allows the photochemical and non-photochemical components of fluorescence quenching, to be estimated. The theory and design of this apparatus is described in detail. The use of the apparatus in the field is demonstrated through a study of the photosynthetic performance of a maize and bean crop during the growing season and by analysis of the photosynthetic performance of crops subjected to nitrogen-stress and a herbicide treatment.     

An Evaluation of the Effects of Ethylene on Carbon Assimilation in Lycopersicon esculentum Mill

Leaf and whole plant gas exchange rates of Lycopersicon esculentumMill, were studied during several days of continuous exposureto ethylene. Steady-state photosynthesis and transpiration ratesof control and ethylene-treated individual leaves were equivalent.However, the photosynthesis and transpiration rates of treatedleaves required at least five times longer to reach 50% of thesteady-state rate. This induction lag was attributed to ethylene—inducedleaf epinasty and temporary acclimation to lower incident lightlevels immediately prior to measurement of gas exchange. Thewhole plant net carbon exchange rate (NCER) of a representativetreated plant was also reduced by 51% after 24 h exposure toethylene relative to both its pre-treatment rate and that ofthe control. Ethylene exposure reduced the growth rate of thetreated plant by 50% when expressed as carbon (C) gain. Theinhibition of NCER and growth rate associated with epinastywas completely reversed when the epinastic leaves were returnedto their original positions and light interception was re-established.The results demonstrate that the inhibition of whole plant CO₂assimilation is indirect and due to reduced light interceptionby epinastic leaves. Morphological changes caused by environmentalethylene are thus shown to reduce plant C accumulation withoutinhibiting leaf photosynthesis processes per se. Key words: Ethylene, carbon assimilation, growth     

Changing Sink Demand Affects the Area but not the Specific Activity of Assimilate Sources in Cantaloupe (Cucumis meloL.)

To better understand source-sink interactions, this work focusedon the influence of fruit number on leaf area and photosyntheticactivity in cantaloupe. To this end, flowers were removed over2 years on two Charentais cultivars to obtain single-fruit plantsand plants with an unrestricted fruit load (which set two tofive fruits and constituted control plants). At the whole plantscale, net photosynthesis was reduced by about 30% under highfruit load. At the leaf scale, a submodel of stomatal conductancewas fitted to the data and was included in a rectangular hyperbolamodel of leaf photosynthesis. Maximum leaf net photosynthesisaveraged 14.83 µmol CO₂m^-2s^-1at 1000 µmol quantam^-2s^-1. Light use efficiency was not affected by fruit loadand equalled 0.040 mol CO₂mol^-1quanta. Leaf area of plants withunrestricted fruit load decreased after 24 days from pollination,while the leaf area of single-fruit plants was still increasing.The decrease was due to production of fewer new leaves per day,whereas the number of senescent leaves and the size of individualleaves were not affected by the treatment. Under high fruitload, cultivar Galoubet developed a larger projected leaf areathan cultivar Talma. Thus it is concluded that: (1) large cantaloupefruits may divert a large amount of assimilates away from, andgrow at the expense of, the canopy; and (2) photosynthesis ofthe canopy was lowered because leaf area was reduced whereasphotosynthetic rate of leaves was not altered.Copyright 1998Annals of Botany Company. Cucumis meloL., fruit load, source-sink interactions, leaf photosynthesis, canopy photosynthesis, leaf area, SLA, source strength.     

Separation of Direct and Indirect Responses of Stomata to Light: Results from a Leaf Inversion Experiment at Constant Intercellular CO2 Molar Fraction

Previous work has shown that stomata respond directly to light,but it was not clear whether the only additional response isthrough CO₂, or whether some other metabolite is involved inthis response. Gas exchange experiments were done with normallypositioned and inverted leaves of Hedera helix to investigatethis problem. The macroscopic optical properties of the leavesand their anatomical structure were also studied. These experimentssnowed that there is no need to postulate the existence of amessenger other than CO₂ to explain the indirect response ofstomata to light. The experiments also showed that leaf inversionaffects both stomatal conductance and photosynthesis, and highlightthe difficulties involved in the interpretation of the effectof leaf inversion on stomata when stomatal conductance measurementsare not done concurrently with measurements of CO₂ flux densityand intercellular CO₂ molar fraction Key words: Hedera helix, ivy, gas exchange, leaf inversion, stomatal conductance, light, CO₂ flux density, photosynthesis     

冠层部位和叶龄对红松光合蒸腾特性的影响

利用Li-6400便携式CO2/H2O红外气体分析仪测定了红松不同冠层部位和叶龄针叶的光合蒸腾特性及其环境影响因子.结果表明:冠层部位和叶龄显著地影响最大净光合速率(Pmax)、光饱和点(LSP)、光补偿点(LCP)、表观最大量子效率(α)、蒸腾速率(Tr)和比叶面积(SLA),但对水分利用效率(WUE)影响不显著.随着冠层部位的下降和叶龄的增加,红松针叶的Pmax逐渐下降,其平均值变动在6·55～9·05μmol·m-2·s-1之间.不同冠层部位和叶龄针叶的LSP和LCP的差异很大,以树冠中部针叶对弱光和强光的利用能力最大.Tr随着冠层部位的下降而降低;不同叶龄针叶的Tr在1·37～1·59mmol·m-2·s-1之间变化.不同部位和叶龄红松针叶的Tr和光合有效辐射存在极显著正相关关系(R2=0·967).红松的WUE与净光合速率紧密相关(R2=0·860).随冠层部位的上升和叶龄的增大,红松针叶的SLA递减,分别在6·61～8·41m2·kg-1和6·65～8·38m2·kg-1之间波动.     

Nonstomatal inhibition of photosynthesis by water stress. Reduction in photosynthesis at high transpiration rate without stomatal closure in field-grown tomato

Large underestimates of the limitation to photosynthesis imposed by stomata can occur because of an error in the standard method of calculating average substomatal pressures of carbon dioxide when heterogeneity of those pressures occurs across a leaf surface. Most gas exchange data supposedly indicating nonstomatal inhibition of photosynthesis by water stress could have this error. However, if no stomatal closure occurs, any reduction in photosynthesis must be due to nonstomatal inhibition of photosynthesis. Net carbon dioxide exchange rates and conductances to water vapor were measured under field conditions in upper canopy leaves of tomato plants during two summers in Beltsville, Maryland, USA. Comparisons were made near midday at high irradiance between leaflets in air with the ambient water vapor content and in air with a higher water content. The higher water content, which lowered the leaf to air water vapor pressure difference (VPD), was imposed either one half hour or several hours before measurements of gas exchange. In both seasons, and irrespective of the timing of the imposition of different VPDs, net photosynthesis increased 60% after decreasing the VPD from 3 to 1 kPa. There were no differences in leaf conductance between leaves at different VPDs, thus transpiration rates were threefold higher at 3 than at 1 kPa VPD. It is concluded that nonstomatal inhibition of photosynthesis did occur in these leaves at high transpiration rate.     

农田冬小麦生长和产量对臭氧动态暴露的响应

 评估臭氧(O₃)污染对农田冬小麦生长和产量的影响是污染生态学和生理生态学研究的重要内容之一。该研究运用开顶式气室(OTC),对冬小麦‘ 嘉403’(Triticum aestivum cv. Jia 403)进行了O₃动态暴露的田间原位试验。实验设置过滤空气组(CF)、自然大气组(NF)和两个不同浓度的 O₃动态暴露组(DO100和DO150)。结果表明：1) O₃浓度增加,一方面可以改变灌浆期冬小麦叶片气体交换参数的日变化规律;另一方面引起表观 光量子产额、光饱和点和光补偿点等光响应参数的显著降低。这表明灌浆期叶片光合能力的下降是气孔因素和非气孔因素共同作用的结果。2) O₃暴露可以改变小麦形态特征,如植株变矮、叶片衰老加速、 叶面积变小,并最终导致产量大幅下降。