Remote sensing of heterogeneity in photosynthetic efficiency, electron transport and dissipation of excess light in Populus deltoides stands under ambient and elevated CO2 concentrations, and in a tropical forest canopy, using a new laser-induced fluorescence transient device

Determining the spatial and temporal diversity of photosynthetic processes in forest canopies presents a challenge to the evaluation of biological feedbacks needed for improvement of carbon and climate models. Limited access with portable instrumentation, especially in the outer canopy, makes remote sensing of these processes a priority in experimental ecosystem and climate change research. Here, we describe the application of a new, active, chlorophyll fluorescence measurement system for remote sensing of light use efficiency, based on analysis of laser‐induced fluorescence transients (LIFT). We used mature stands of Populus grown at ambient (380 ppm) and elevated CO₂ (1220 ppm) in the enclosed agriforests of the Biosphere 2 Laboratory (B2L) to compare parameters of photosynthetic efficiency, photosynthetic electron transport, and dissipation of excess light measured by LIFT and by standard on‐the‐leaf saturating flash methods using a commercially available pulse‐modulated chlorophyll fluorescence instrument (Mini‐PAM). We also used LIFT to observe the diel courses of these parameters in leaves of two tropical forest dominants, Inga and Pterocarpus, growing in the enclosed model tropical forest of B2L. Midcanopy leaves of both trees showed the expected relationships among chlorophyll fluorescence‐derived photosynthetic parameters in response to sun exposure, but, unusually, both displayed an afternoon increase in nonphotochemical quenching in the shade, which was ascribed to reversible inhibition of photosynthesis at high leaf temperatures in the enclosed canopy. Inga generally showed higher rates of photosynthetic electron transport, but greater afternoon reduction in photosynthetic efficiency. The potential for estimation of the contribution of outer canopy photosynthesis to forest CO₂ assimilation, and assessment of its response to environmental stress using remote sensing devices such as LIFT, is briefly discussed.     

Variation of photosynthetic capacity with leaf age in an alpine orchid, Cypripedium flavum

Photosynthetic rate, chlorophyll fluorescence, leaf nitrogen and chlorophyll content of Cypripedium flavum were studied at different leaf ages. The photosynthetic capacity changed significantly with leaf age. Net photosynthesis and chlorophyll content peaked when leaf age was 60 days, decreasing at 30, 90 and 120 days. Stomatal conductance showed the highest value at 60 days, while mesophyll conductance decreased with increasing leaf age. Both leaf nitrogen content per unit area and leaf nitrogen content per unit mass decreased with increasing leaf age. The age-dependent variation in photosynthetic capacity could be linked to the changes in biochemical efficiency, leaf nitrogen content and CO₂ diffusion limitation.     

Changes in chlorophyll a fluorescence, photosynthetic CO2 assimilation and xanthophyll cycle interconversions during dehydration in desiccation-tolerant and intolerant liverworts

The interactions among water content, chlorophyll a fluorescence emission, xanthophyll interconversions and net photosynthesis were analyzed during dehydration in desiccation-tolerant Frullania dilatata (L.) Dum. and desiccation-intolerant Pellia endiviifolia (Dicks) Dum. Water loss led to a progressive suppression of photosynthetic carbon assimilation in both species. Their chlorophyll fluorescence characteristics at low water content were: low photosynthetic quantum conversion efficiency, high excitation pressure on photosystem II and strong non-photochemical quenching. However, dissipation activity was lower in P. endiviifolia and was not accompanied by a rise in the concentration of de-epoxidised xanthophylls as F. dilatata. The photosynthetic apparatus of F. dilatata remained fully and speedily recuperable after desiccation in as indicated by the restoration of chlorophyll fluorescence parameters to pre-desiccation values upon rehydration. A lack of recovery upon remoistening of P. endiviifolia indicated permanent and irreversible damage to photosystem II. The results suggest that F. dilatata possesses a desiccation-induced zeaxanthin-mediated photoprotective mechanism which might aid photosynthesis recovery when favourable conditions are restored by alleviating photoinhibitory damage during desiccation. This avoidance mechanism might have evolved as an adaptative response to repeated cycles of desiccation and rehydration that represent a real threat to photosynthetic viability. Received: 12 January 1998 / Accepted: 14 July 1998     

Response of Tetraselmis suecica to nutrient and grazer manipulation

Three methods of algal quantification (direct cell counts, chlorophyll a extraction, in vivo fluorescence) were used to evaluate the response of the unicellular green flagellate Tetraselmis suecica to nutrients and grazers. Nutrient enrichment enhanced total cell counts, chlorophyll a concentration and in vivo and DCMU-fluorescence. Photosynthetic efficiency was reduced in the complete F2 medium as indicated by the high level of in vivo fluorescence, whereas photosynthetic efficiency was increased by the introduction of mussels to the F2 medium. The addition of mussels significantly increased the proportion of non-motile cells, but did not reduce the total cell count. The effect of mussel grazing on algae could be underestimated if only total cells were counted or only the chlorophyll a concentration was measured. The results indicate that these three methods measure different properties of an algal culture and are complementary to each other in assessing the quality and quantity of an algal population. Direct algal counting offers a reliable numerical assessment for cell population abundance. Chlorophyll a concentration was closely correlated to the total cell count. In the presence of mussels, in vivo fluorescence did not correlate with either algal cell counts or chlorophyll a concentration, indicating that the measurement of in vivo fluorescence may be misleading for estimating algal abundance under different culture conditions. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effect of transient accumulation of anthocyanin on leaf development and photoprotection of Fagopyrum dibotrys mutant

Changes in pigments contents, leaf area, leaf dry mass per unit area (LMA), photosynthetic rate and chlorophyll a fluorescence were investigated in developing leaves of Fagopyrum dibotrys Hara. mutant. Anthocyanins transiently accumulate below the upper epidermis during leaf ontogeny of this mutant. Red leaves possessed lower Chl content, LMA, photosynthetic rate and apparent carboxylation efficiency than green leaves. However, content of anthocyanins declined and above mentioned parameters increased during further leaf development. In both red and green leaves, chronic photoinhibition did not take place according to variable to maximum chlorophyll fluorescence ratio (F_v/F_m). Red leaves had higher non-photochemical quenching (NPQ) and higher PS 2 efficiency.     

银杏叶萜内酯含量的变化及其与叶绿素荧光特性的关系

为探究银杏(Ginkgo biloba)叶萜类内酯含量和光合同化作用的关系,对其内酯含量和叶绿素荧光特性进行了研究。结果表明,不同采收时间银杏叶中白果内酯和银杏内酯含量有显著差异,总体上,5月份含量较低,此后逐渐升高,8月份达到高峰,然后快速下降,10月底最低;与此同时,银杏叶片的光合色素以及叶绿素荧光参数也呈现周期性变化。白果内酯以及萜内酯含量与叶绿素荧光参数Y(NPQ)之间呈极显著正相关关系,因此,可以通过银杏叶片的叶绿素荧光参数预测白果内酯和萜内酯含量。     

Oscillations in photosynthetic carbon assimilation and chlorophyll fluorescence are different in Amaranthus caudatus,a C4 plant,and Spinacia oleracea,a C3 plant

The characteristics of oscillations in photosynthetic carbon fixation and chlorophyll fluorescence in leaves of the C₄ plant Amaranthus caudatus L. were compared to those shown by the C₃ plant Spinacia oleracea L. As in spinach, oscillations could be observed in Amaranthus when leaves were illuminated after periods of darkening, particularly at temperatures below 20°C, less so or not at all at higher temperatures. However, in contrast to spinach, pronounced oscillations occurred in Amaranthus after a sudden dark/light transition only at low, not at high photon flux densities. Whereas in spinach maxima in carbon uptake were observed slightly after minima in chlorophyll fluorescence had occurred, in Amaranthus maxima in carbon uptake were close to maxima in chlorophyll fluorescence. Since the quantum efficiency of electron transport through photosystem II of the chloroplast electron-transport chain was higher during the minima of chlorophyll fluorescence than during the maxima, the observations suggest that in Amaranthus photosynthetic water oxidation did not occur as synchronously with carbon uptake as in spinach. It is proposed that, in contrast to spinach, photosynthetic oscillations in Amaranthus are related to the diffusional transport of photosynthetic intermediates between mesophyll and bundle-sheath cells.Abbreviations F_o, F_m, F_s initial, maximal and steady-state chlorophyll a fluorescence - PFD photon flux density - Q_A primary quinone acceptor of PSII We are grateful to Professors D.A. Walker, FRS, Robert Hill Institute, University of Sheffield, Sheffield, UK., and Agu Laisk, Chair of Plant Physiology, University of Tartu, Tartu, Estonia, for helpful discussions and to Ms. S. Neimanis for help with the experiments. Our work was performed within the research of the Sonderforschungsbereich 251 of the University of Würzburg. It was supported by the Stiftung Volkswagenwerk. A.S.R. acknowledges also support by the Alexander-von-Humboldt-Stiftung and U.G. by the Graduate College of the University of Würzburg.     

The usefulness of the chlorophyll fluorescence parameters in harvest prediction in 10 genotypes of winter triticale under optimal growth conditions

The aim of the experiment was to detect differences in the activity of the photosynthetic apparatus in 10 genotypes of winter triticale. Measurements of gas exchange and chlorophyll fluorescence were performed. Among the tested genotypes, the photosynthetic apparatus of Timbo and Piano was the most active, while the photosynthesis of both Babor and Boreas was highly reduced. Additionally, we have found significant correlations between the yield and some parameters of chlorophyll fluorescence and leaf gas exchange. Parameters of chlorophyll fluorescence are useful for estimation of the functional state of the photosynthetic apparatus and could become selection criteria in plant breeding. Moreover, such parameters permit definition of the effectiveness of the utilization of chlorophyll a excitation energy and co‐operation of the light phase reactions, with reactions occurring during the dark phase of photosynthesis.     

Photosynthesis impairment in cassava leaves in response to nitrogen deficiency

Plants of cassava (Manihot esculenta Crantz cv. Cigana Preta) grown in a sand root medium were watered with nutrient solutions containing either 3 mM nitrate (low N) or 12 mM nitrate (high N). Chlorophyll concentration, chlorophyll a/b ratio, stomatal conductance, photorespiration rate and net carbon assimilation rate (on an area and a mass basis, but not on a chlorophyll basis) all decreased in low-N plants as compared with high-N ones. By contrast, photosynthetic nitrogen-use efficiency increased in low-N plants. As indicated by chlorophyll a fluorescence data, these plants exhibited increases in both excitation pressure on Photosystem II and thermal energy dissipation, with a corresponding decrease in quantum yield of electron transport, when contrasted with high-N plants. This decrease paralleled an unchanged maximal Photosystem II photochemical efficiency, suggesting a down-regulation of the Photosystem II photochemistry. It is proposed that decline in biochemical capacity for carboxylation, rather than stomatal limitation or electron transport, were the major constraints associated to the reduced photosynthetic rates induced by nitrogen deficiency in cassava plants.     

西南桦幼苗接种丛枝菌根真菌的生长与光合生理响应

为了解丛枝菌根真菌(arbuscular mycorrhiza,AMF)对西南桦幼苗生长和光合生理的影响,对西南桦(Betula alnoides)优良无性系接种AMF菌株后的生长、光合参数、叶绿素含量和荧光参数进行了研究。结果表明,6个AMF菌株均能与西南桦无性系幼苗形成共生体,接种根内球囊霉(Glomus intraradices)菌株(AMF5)和摩西球囊霉(G.mosseae)HUN03B菌株(AMF3)显著提高了幼苗生长量、净光合速率、水分利用效率、叶绿素含量和荧光参数(P0.05),显示出AMF5、AMF3与幼苗的亲和力明显优于其他菌株。西南桦4个无性系间的菌根侵染率差异不显著(P0.05),但AMF对无性系FB4、BY1的促生效应显著优于FB4+和A5。因此,适合与西南桦共生的优良菌株为AMF5和AMF3,这为西南桦菌根化育苗提供理论依据。     

Chloroplast ultrastructure and fluorescence response of oilseed rape containing male-sterile radish cytoplasm

Summary The production of hybrid seed is facilitated if one parent possesses a male-sterile cytoplasm. Introduction of the cytoplasm of male-sterile radish (Raphanus sativus L.) into rapeseed (Brassica napus L.) results not only in transfer of the desirable male-sterile trait but induces a chlorophyll defect in the backcrossed male-sterile plants. In this study we show that the defect manifests itself in two different ways in the alloplasmic plants: a) smaller and fewer chloroplasts with an impaired ultrastructure and b) an increase in chlorophyll fluorescence. Defective chloroplasts were characterized by a reduction in both the number and size of grana, the latter due to poor stacking of thylakoids and with frequent discontinuity in the intergranal thylakoid systems. The changed chloroplast morphology and the increase in chlorophyll fluorescence are probably the cause of the lowered photosynthetic efficiency associated with the alloplasmic plants. We propose that the deficiency is the result of incompatibility between the genomes of the radish chloroplast and the rapeseed nucleus. Supporting this hypothesis are studies of male-sterile rapeseed plants in which, by protoplast fusion, the radish chloroplasts were substituted by those of normal male-fertile rapeseed. Such plants showed complete restoration of their photosynthetic potential and displayed both normal chloroplast ultrastructure and normal levels of chlorophyll fluorescence.     

Diverse Optical and Photosynthetic Properties in a Neotropical Dry Forest during the Dry Season: Implications for Remote Estimation of Photosynthesis1

Using optical and photosynthetic assays from a canopy access crane, we examined the photosynthetic performance of tropical dry forest canopies during the dry season in Parque Metropolitano, Panama City, Panama. Photosynthetic gas exchange, chlorophyll fluorescence, and three indices derived from spectral reflectance (the normalized difference vegetation index, the simple ratio, and the photochemical reflectance index) were used as indicators of structural and physiological components of photosynthetic activity. Considerable interspecific variation was evident in structural and physiological behavior in this forest stand, which included varying degrees of foliage loss, altered leaf orientation, stomatal closure, and photosystem II downregulation. The normalized difference vegetation index and the simple ratio were closely related to canopy structure and absorbed radiation for most species, but failed to capture the widely divergent photosynthetic behavior among evergreen species exhibiting various degrees of downregulation. The photochemical reflectance index and chlorophyll fluorescence were related indicators of photosynthetic downregulation, which was not detectable with the normalized difference vegetation index or simple ratio. These results suggest that remote sensing methods that ignore downregulation cannot capture within‐stand variability in actual carbon flux for this diverse forest type. Instead, these findings support a sampling approach that derives photosynthetic fluxes from a consideration of both canopy light absorption (e.g., normalized difference vegetation index) and photosynthetic light‐use efficiency (e.g., photochemical reflectance index). Such sampling should improve our understanding of controls on photosynthetic carbon uptake in diverse tropical forest stands.     

Influence of bacterial leaf blight on the photosynthetic characteristics of resistant and susceptible rice

The effect of crop disease on photosynthetic characteristics is important for disease control. Two varieties, Shenzhou 98 and Neiwuyou 8015 with resistance and susceptibility to bacterial leaf blight (BLB), respectively, were selected, and the responses of the net photosynthetic rate (P_N) to active photon flux density (PPFD) and intercellular carbon dioxide concentration (C_i), as well as chlorophyll fluorescence, pigments and stomatal resistance (SR), were measured. The results showed that BLB infection greatly decreased the maximum photosynthetic rate (P_max), light saturation point (LSP), carboxylation efficiency (CE), maximal fluorescence (F_m) and actual photochemical efficiency of PSII ( Φ _PSII) but increased the light compensation point (LCP) and dark respiratory rate (R_D), which suggested that the performance of rice photosynthesis was decreased by BLB infection. The BLB infection had a lower effect on resistant rice Shenzhou 98 than on susceptible rice Neiwuyou 8015. The reduction of pigment and increased SR caused by BLB infection may have resulted in the decline in the photosynthetic rate. Significant effects of the BLB infection were observed on chlorophyll fluorescence F_m and Φ_PSII in resistant and susceptible rice. These parameters may be useful for noninvasive monitoring of plant disease considering the negative effect caused by other stresses.     

Chlorophyll a fluorescence: can it shed light on fundamental questions in photosynthetic carbon dioxide fixation?

Abstract A close, immediate and precise relationship between chlorophyll a fluorescence and photosynthetic carbon assimilation in vivo is demonstrated. The examples discussed include kinetics displayed during dark to light transitions plus oscillations and transients observed during changes in the gas phase surrounding the leaf. Remaining uncertainties surrounding the relationship between chlorophyll fluorescence and photosynthesis are attributed to the underlying complexity of the regulatory mechanisms involved. Examples are also given that show how multiple simultaneous measurements of different aspects of the photosynthetic process may contribute to the resolution of these uncertainties. The practical relevance of these matters is also discussed, particularly in relation to the limitations of the photosynthetic process and to the use of chlorophyll fluorescence as a diagnostic probe of chemical and genetic manipulation and stress.     

Glucose feeding results in coordinated changes of chlorophyll content, ribulose-1,5-bisphosphate carboxylase-oxygenase activity and photosynthetic potential in photoautrophic suspension cultured cells of Chenopodium rubrum

The effects of an excessive supply of carbohydrates on the photosynthetic apparatus of photoautotrophic suspension cultured cells of Chenopodium rubrum were analysed. Glucose feeding resulted in a massive intracellular accumulation of carbohydrates (glucose, fructose, sucrose and starch) and a delayed but pronounced reduction of the chlorophyll content. Maximum photosynthetic efficiency (determined as relative photon yield of oxygen evolution or calculated from fluorescence parameters) was only slightly affected by this treatment. The maximum values of photosynthetic oxygen evolution and of ribulose-1,5-bisphosphate carboxylase-oxygenase activity also did not change significantly when expressed on a chlorophyll basis, thus indicating that membrane and stroma components were reduced to the same extent. Growth analysis demonstrated that even with an excessive supply of carbohydrates the contribution of photosynthesis to growth was still significant (about 35% of total DW production). The results support the hypothesis that accumulation of carbohydrates leads to a coordinated reduction of the photosynthetic machinery. Comparable changes possibly occur during source/sink transitions in green plant tissues.     

17个建兰品种光合特性分析

以17个建兰(Cymbidium ensifolium)品种为材料,采用改良的丙酮法提取叶绿素,再通过Arnon丙酮法公式计算光合色素含量,利用捷克FluorCam开放式叶绿素荧光仪测定不同品种的叶绿素荧光参数。结果表明,17个建兰品种的光合色素和叶绿素荧光参数具有不同程度的差异,其中‘铁骨素’(C. ensifolium ‘Tiegusu’)、‘逸红双娇’(C. ensifolium ‘Yihongshuangjiao’)和‘闽南黄蝶’(C. ensifolium ‘Minnanhuangdie’)的光合色素含量高于其他品种,表明这3个品种具有良好的光合效率,吸收光能的能力较强;‘铁骨素’最大荧光产量(Fm)、Kautsky诱导效应最大荧光(Fp)、PS Ⅱ原初光能转化效率(Fv/Fm)和非光化荧光淬灭系数(NPQ)均为最高。综上可知,‘铁骨素’的光合生理特性优于其他品种,可作为优良建兰品种进行种植推广。     

不同光强对越南抱茎茶光合及荧光特性日变化的影响

为研究不同光强对越南抱茎茶光合作用机理的影响,设置3种处理方式,即全光照(L1)、光强度45%(L2)、光强度15%(L3),分别在遮阴3个月后测定其光合荧光参数的变化。结果表明,不同光强处理下,净光合速率均呈单峰曲线,峰值出现在11:00左右;光合速率下降主要是气孔限制因素引起的;叶绿素荧光参数表明,强光条件下电子传递受阻,热耗散增加,PSII反应中心受到破坏,从而降低了茶花的光合效率;过度遮阴条件下受到光抑制,从而导致光合机构损伤;45%光照条件下光合效率较高。相关性分析表明,净光合速率(Pn)与空气温度(Ta)、相对湿度(RH)、气孔导度(Gs)、蒸腾速率(Tr)、水分利用效率(WUE)、表观电子传递速率(ETR)呈极显著相关,与RH呈极显著负相关,与胞间CO2浓度(Ci)呈显著负相关。影响越南抱茎茶净光合速率的主要环境因子是空气温度和相对湿度。     

In situ estimation of net CO2 assimilation, photosynthetic electron flow and photorespiration in Turkey oak (Q. cerris L.) leaves: diurnal cycles under different levels of water supply

Diurnal time courses of net CO₂ assimilation rates, stomatal conductance and light-driven electron fluxes were measured in situ on attached leaves of 30-year-old Turkey oak trees (Quercus cerris L.) under natural summer conditions in central Italy. Combined measurements of gas exchange and chlorophyll a fluorescence under low O₂ concentrations allowed the demonstration of a linear relationship between the photochemical efficiency of PSII (fluorescence measurements) and the apparent quantum yield of gross photosynthesis (gas exchange). This relationship was used under normal O₂ to compute total light-driven electron fluxes, and to partition them into fractions used for RuBP carboxylation or RuBP oxygenation. This procedure also yielded an indirect estimate of the rate of photorespiration in vivo. The time courses of light-driven electron flow, net CO₂ assimilation and photorespiration paralleled that of photosynthetic photon flux density, with important afternoon deviations as soon as a severe drought stress occurred, whereas photochemical efficiency and maximal fluorescence underwent large but reversible diurnal decreases. The latter observation indicated the occurrence of a large non-photochemical energy dissipation at PSII. We estimated that less than 60% of the total photosynthetic electron flow was used for carbon assimilation at midday, while about 40% was devoted to photorespiration. The rate of carbon loss by photorespiration (R₁) reached mean levels of 56% of net assimilation rates. The potential application of this technique to analysis of the relative contributions of thermal de-excitation at PSII and photorespiratory carbon recycling in the protection of photosynthesis against stress effects is discussed.     

Seasonal and diurnal monitoring of leaf polyamine content in Quercus ilex L. resprouts after fire in relation to changes in irradiation and photosynthetic parameters

Seasonal variations in free putrescine, spermidine and spermine content, gas-exchange and chlorophyll fluorescence parameters were followed during winter and summer on leaves of a similar age from undisturbed holm oak trees (control, C) and resprouts (R) originated after fire. We observed a general trend of putrescine content decrease with increasing irradiance. Putrescine content decreased markedly from winter to summer, especially in R, which were located on a site with much higher irradiation. Daily summer variations in putrescine showed a decline at midday from morning values, and they were also more accentuated in R. Measurement of gas-exchange and chlorophyll fluorescence parameters showed marked differences between C and R under their respective light conditions. R showed higher values of PSII quantum yield (Φ_PSII), photochemical quenching (qP) and intrinsic efficiency of open PSII centres () The Φ_PSII/PPFD response curve showed that under the same irradiance, Φ_PSII was enhanced in R and mainly under high light conditions. In spite of increasing irradiance from winter to summer, and especially in burned areas, the mentioned chlorophyll fluorescence parameters were maintained indicating the adaptation of the photosynthetic apparatus. Results derived from A/C _i and A/PPFD response curves showed enhanced photosynthetic capacity and lower non-stomatal limitation of photosynthesis in R during summer stress. The contribution of putrescine decline in the photoadaptation of the photosynthetic apparatus of species growing in natural forest habitats is considered.     

Analysis of Photosynthetic Activity at the Leaf and Canopy Levels from Reflectance Measurements: A Case Study

The photochemical reflectance index (PRI), based on reflectance signatures at 531 and 570 nm, and associated with xanthophyll pigment inter-conversion and related thylakoid energisation, was evaluated as an indicator of photosynthetic function in a Mediterranean holm oak (Quercus ilex L.) coppice. The chlorophyll fluorescence pulse-amplitude-modulation and the eddy correlation techniques were used to estimate the photosystem 2 photochemical efficiency of leaves and the CO₂ flux over the canopy, respectively. The reflectance and fluorescence techniques yielded identical estimates of the photosynthetic activity in leaves exposed to dark-light-dark cycles or to a variable irradiance in laboratory. However, there was no such correlation between photosynthetic performance and PRI when applied to a sun-exposed canopy in field conditions. Fluorescence profiles inside the canopy and especially a helpful use of multispectral reflectance imaging highlight the limitations of such method.