Chlorophyll a fluorescence and light-scattering kinetics displayed by leaves during induction of photosynthesis

Light-scattering, which can be taken as an indicator of the transthylakoid proton-gradient, and chlorophyll a fluorescence, have been followed simultaneously during re-illumination of spinach leaves at different energy fluence rates and carbon dioxide concentrations. The slow fluorescence transient (M peak), which has been associated with photosynthetic induction, was observed in air only at the lower fluence rates used. Data are presented that indicate that M peaks in chlorophyll fluorescence kinetics can only be observed if there is also a simultaneous transient in light-scattering and that these transients are observed when the dark period is relatively long, fluence rate relatively low, and CO₂ concentration relatively high.The results are discussed in relation to the varying demands on ATP by carbon assimilation during induction of photosynthesis at different carbon dioxide concentrations and the manner in which these variations influence the quenching of chlorophyll a fluorescence.Abbreviation Chl chlorophyll     

Chlorophyll fluorescence and photoinhibition in a tropical rainforest understory plant

The data presented here deal with the effects of high-light exposure on the 77 K fluorescence characteristics of Elatostema repens. It is shown that the decrease of the variable fluorescence during the treatment is biphasic. The reactions responsible for the first phase of fluorescence quenching are saturated under 700 mol photon m^-2 s^-1 and insensitive to streptomycin, whereas those responsible for the second phase are not yet saturated under 700 mol photon m^-2 s^-1 and sensitive to streptomycin. It is concluded that only the second phase of fluorescence quenching is associated with photoinhibitory processes. Rate and amplitude of recovery from photoinhibition are maximum under very low light (3.5 mol photon m^-2 s^-1), and very small at a moderate light (160 mol photon m^-2 s^-1) which does not cause photoinhibition. It is concluded that recovery processes are inhibited during photoinhibition. It is suggested that they could be associated with damage occuring on the oxidizing side of PSII.Abbreviations F_o, F_v, F_m initial, variable and maximum fluorescence, respectively - PFD photon flux density - PS II photosystem II     

Chlorophyll fluorescence tracks seasonal variations of photosynthesis from leaf to canopy in a temperate forest

Accurate estimation of terrestrial gross primary productivity (GPP) remains a challenge despite its importance in the global carbon cycle. Chlorophyll fluorescence (ChlF) has been recently adopted to understand photosynthesis and its response to the environment, particularly with remote sensing data. However, it remains unclear how ChlF and photosynthesis are linked at different spatial scales across the growing season. We examined seasonal relationships between ChlF and photosynthesis at the leaf, canopy, and ecosystem scales and explored how leaf‐level ChlF was linked with canopy‐scale solar‐induced chlorophyll fluorescence (SIF) in a temperate deciduous forest at Harvard Forest, Massachusetts, USA. Our results show that ChlF captured the seasonal variations of photosynthesis with significant linear relationships between ChlF and photosynthesis across the growing season over different spatial scales (R² = 0.73, 0.77, and 0.86 at leaf, canopy, and satellite scales, respectively; P < 0.0001). We developed a model to estimate GPP from the tower‐based measurement of SIF and leaf‐level ChlF parameters. The estimation of GPP from this model agreed well with flux tower observations of GPP (R² = 0.68; P < 0.0001), demonstrating the potential of SIF for modeling GPP. At the leaf scale, we found that leaf F_q’/F_m’, the fraction of absorbed photons that are used for photochemistry for a light‐adapted measurement from a pulse amplitude modulation fluorometer, was the best leaf fluorescence parameter to correlate with canopy SIF yield (SIF/APAR, R² = 0.79; P < 0.0001). We also found that canopy SIF and SIF‐derived GPP (GPP_SIF) were strongly correlated to leaf‐level biochemistry and canopy structure, including chlorophyll content (R² = 0.65 for canopy GPP_SIF and chlorophyll content; P < 0.0001), leaf area index (LAI) (R² = 0.35 for canopy GPP_SIF and LAI; P < 0.0001), and normalized difference vegetation index (NDVI) (R² = 0.36 for canopy GPP_SIF and NDVI; P < 0.0001). Our results suggest that ChlF can be a powerful tool to track photosynthetic rates at leaf, canopy, and ecosystem scales.     

Heat-induced electrical signals affect cytoplasmic and apoplastic pH as well as photosynthesis during propagation through the maize leaf

Combining measurements of electric potential and pH with such of chlorophyll fluorescence and leaf gas exchange showed heat stimulation to evoke an electrical signal (propagation speed: 3–5 mm s⁻¹) that travelled through the leaf while reducing the net CO₂ uptake rate and the photochemical quantum yield of both photosystems (PS). Two-dimensional imaging analysis of the chlorophyll fluorescence signal of PS II revealed that the yield reduction spread basipetally via the veins through the leaf at a speed of 1.6 ± 0.3 mm s⁻¹ while the propagation speed in the intervein region was c. 50 times slower. Propagation of the signal through the veins was confirmed because PS I, which is present in the bundle sheath cells around the leaf vessels, was affected first. Hence, spreading of the signal along the veins represents a path with higher travelling speed than within the intervein region of the leaf lamina. Upon the electrical signal, cytoplasmic pH decreased transiently from 7.0 to 6.4, while apoplastic pH increased transiently from 4.5 to 5.2. Moreover, photochemical quantum yield of isolated chloroplasts was strongly affected by pH changes in the surrounding medium, indicating a putative direct influence of electrical signalling via changes of cytosolic pH on leaf photosynthesis.     

Chlorophyll a fluorescence during flag leaf senescence of field-grown winter wheat plants under drought conditions

Improving wheat grain yield plays a significant role in ensuring global food security. Wheat production could be increased by the genetic improvement of wheat genotypes where delayed senescence with enhanced post-anthesis capacity and staygreen traits could have an important role. In this study, chlorophyll a fluorescence (ChlF) rise kinetics from the early until late senescence of flag leaves, grain yield and other agro-morphological characteristics were compared for three winter wheat advanced lines (Osk.4.312/10-18, Osk.4.330/6-18 and Osk.4.354/12-18) under natural drought conditions. The differences between lines were observed when considering the heading date which was 1 and 4 days earlier for the line Osk.4.354/12-18, than lines Osk.4.312/10-18 and Osk.4.330/6-18, respectively. Furthermore, line Osk.4.354/12-18 had the highest test weight (kg hl⁻¹), while line Osk.4.330/6-18 showed a tendency of decreased grain yield, compared to the other two lines. Analysis of ChlF transients and several JIP-test parameters indicated that all three lines had a generally similar course of changes in the photosynthetic performance of flag leaves during senescence under drought conditions. However, at the point when a decrease in photosynthetic performance was initiated, it was slightly less intensive in line Osk.4.354/12-18 accompanied by longer preservation of functionality and connectivity of PSII units, than in the other two lines, which contributed to its better agronomical performance. These results indicated that even delicate variations in the functioning of the photosynthetic apparatus of the flag leaf during grain filling were agronomically important, especially when plants were exposed to drought stress, and could be used to differentiate otherwise similar wheat genotypes. Even small genotype-specific differences in the photosynthetic performance of senescing flag leaves, along with earlier heading dates, could assist in the selection of genotypes with a better ability to cope with unfavourable environmental conditions.     

Chlorophyll fluorescence quenching in the alga Euglena gracilis

When far red light preincubated cells of Euglena gracilis are transferred to dark or light, chlorophyll fluorescence (F₀ and F_m) decreases. Non-photochemical quenching in the dark is suggested to be induced partly by chlororespiration and partly by changes in the distribution of excitation energy between the photosystems. Depending on the light intensities it was possible to resolve the non-photochemical quenching into at least three different components. The slowest relaxation phase of non-photochemical quenching occurred only after exposure to high light and was assigned to photoinhibition. The other two components were an energy-dependent quenching (qE), and the one which we attribute to a spill over mechanism. We suggest that both photosystems use a common antenna system consisting of LHC I and LHC II proteins. In contrast to higher plants, qE in Euglena gracilis is independent of the xanthophyll cycle and an aggregation of LHC II. This revised version was published online in June 2006 with corrections to the Cover Date.     

The photosynthesis of individual algal cells during the cell cycle of Scenedesmus quadricauda studied by chlorophyll fluorescence kinetic microscopy

A microscope for imaging of chlorophyll fluorescence kinetics was equipped with a chamber that allows the growth of an immobilised population of algae and their study under well-defined conditions. Single cells of the chlorococcal alga Scenedesmus quadricauda were grown and recorded for periods of whole cell cycles (up to 48 h) displaying a normal course of cell development. Heterogeneity in fluorescence yield among individual coenobia in the population and among different cells in one coenobium were analysed. Differences were observed both in the shape of Kautsky transients and in the modulation of fluorescence parameter values during the progress of the cell cycle. The extent of heterogeneity in fluorescence parameters was cell cycle dependent – in some phases of the cycle, the population was almost homogeneous, while distinct heterogeneity was observed, in particular between the protoplast division and the release of the daughter coenobia. The heterogeneity was not random but reflected developmental processes.     

Diurnal changes in leaf gas exchange and chlorophyll fluorescence in tropical tree species with contrasting light requirements

Interspecific ecophysiological differences in response to different light environments are important to consider in regeneration behavior and forest dynamics. The diurnal changes in leaf gas exchange and chlorophyll fluorescence of two dipterocarps, Shorea leprosula (a high light-requiring) and Neobalanocarpus heimii (a low light-requiring), and a pioneer tree species (Macaranga gigantea) growing in open and gap sites were examined. In the open site, the maximum net photosynthetic rate (P_n), photosystem II (PSII) quantum yield (; F/Fm), and relative electron transport rate (r-ETR) through PSII at a given photosynthetic photon flux density (PPFD) was higher in S. leprosula and M. gigantea than in N. heimii, while non-photochemical quenching (NPQ) at a given PPFD was higher in N. heimii. The maximum values of net photosynthetic rate (P_n) in M. gigantea and S. leprosula was higher in the open site (8–11 mol m^–2 s^–1) than in the gap site (5 mol m^–2 s^–1), whereas that in N. heimii was lower in the open site (2 mol m^–2 s^–1) than in the gap site (4 mol m^–2 s^–1), indicating that N. heimii was less favorable to the open site. These data provide evidence to support the hypothesis that ecophysiological characteristics link with plants regeneration behavior and successional status. Although P_n and stomatal conductance decreased at midday in M. gigantea and S. leprosula in the open site, both r-ETR and leaf temperature remained unchanged. This indicates that stomatal closure rather than reduced photochemical capacity limited P_n in the daytime. Conversely, there was reduced r-ETR under high PPFD conditions in N. heimii in the open site, indicating reduced photochemical capacity. In the gap site, P_n increased in all leaves in the morning before exposure to direct sunlight, suggesting a relatively high use of diffuse light in the morning.     

田野菟丝子寄生对五种红树植物叶绿素荧光参数的影响

红树林由于受到入侵植物薇甘菊的危害发生大面积退化,而田野菟丝子寄生是防控薇甘菊的有效途径。为了探讨田野莬丝子寄生对红树植物的影响,该研究通过同质园控制试验,利用PAM-2100便携式调制叶绿素荧光仪,测定了三种真红树植物(老鼠簕、木榄、秋茄)和两种半红树植物(海芒果、银叶树)幼苗在田野菟丝子寄生及对照情况下的叶片叶绿素荧光参数。结果表明:田野菟丝子寄生对红树植物幼苗光系统Ⅱ(PSⅡ)最大光化学效率(F_v/F_m)没有显著影响;对更为敏感的红树植物幼苗PSⅡ潜在活性(F_v/F_o)同样也没有显著影响;试验后期发现田野菟丝子对红树植物幼苗无法成功寄生。由此可见,田野菟丝子用于红树林薇甘菊的防治是安全的。     

Effect of Polyamines on Chlorophyll and Protein Contents, Photochemical Activity, and Energy Transfer in Detached Wheat Leaves during Dark Incubation

Spermine as compared to putrescine or spermidine retarded the loss of chlorophyll and protein contents to a greater extent in wheat primary leaves during dark incubation. Activities of whole chain electron transport, photosystem (PS) 1 and PS2, and absorbed excitation energy distribution in favour of PS 1 were protected by these amines in valency dependent manner during 72-h dark incubation.     

New fluorescence parameters for monitoring photosynthesis in plants

Chlorophyll fluorescence measurements have a wide range of applications from basic understanding of photosynthesis functioning to plant environmental stress responses and direct assessments of plant health. The measured signal is the fluorescence intensity (expressed in relative units) and the most meaningful data are derived from the time dependent increase in fluorescence intensity achieved upon application of continuous bright light to a previously dark adapted sample. The fluorescence response changes over time and is termed the Kautsky curve or chlorophyll fluorescence transient. Recently, Strasser and Strasser (1995) formulated a group of fluorescence parameters, called the JIP-test, that quantify the stepwise flow of energy through Photosystem II, using input data from the fluorescence transient. The purpose of this study was to establish relationships between the biochemical reactions occurring in PS II and specific JIP-test parameters. This was approached using isolated systems that facilitated the addition of modifying agents, a PS II electron transport inhibitor, an electron acceptor and an uncoupler, whose effects on PS II activity are well documented in the literature. The alteration to PS II activity caused by each of these compounds could then be monitored through the JIP-test parameters and compared and contrasted with the literature. The known alteration in PS II activity of Chenopodium album atrazine resistant and sensitive biotypes was also used to gauge the effectiveness and sensitivity of the JIP-test. The information gained from the in vitro study was successfully applied to an in situ study. This is the first in a series of four papers. It shows that the trapping parameters of the JIP-test were most affected by illumination and that the reduction in trapping had a run-on effect to inhibit electron transport. When irradiance exposure proceeded to photoinhibition, the electron transport probability parameter was greatly reduced and dissipation significantly increased. These results illustrate the advantage of monitoring a number of fluorescence parameters over the use of just one, which is often the case when the F_V/F_M ratio is used. This revised version was published online in August 2006 with corrections to the Cover Date.     

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

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

Chlorophyll fluorescence imaging for disease-resistance screening of sugar beet

Both biotic and abiotic stresses cause considerable crop yield losses worldwide (Chrispeels, Sadava Plants, genes, and crop biotechnology 2003; Oerke, Dehne Crop Prot 23:275–285 2004). To speed up screening assays in stress resistance breeding, non-contact techniques such as chlorophyll fluorescence imaging can be advantageously used in the quantification of stress-inflicted damage. In comparison with visual spectrum images, chlorophyll fluorescence imaging reveals cell death with higher contrast and at earlier time-points. This technique has the potential to automatically quantify stress-inflicted damage during screening applications. From a physiological viewpoint, screening stress-responses using attached plant leaves is the ideal approach. However, leaf growth and circadian movements interfere with time-lapse monitoring of leaves, making it necessary to fix the leaves to be studied. From this viewpoint, a method to visualise the evolution of chlorophyll fluorescence from excised leaf pieces kept in closed petri dishes offers clear advantages. In this study, the plant–fungus interaction sugar beet–Cercospora beticola was assessed both in attached leaf and excised leaf strip assays. The attached leaf assay proved to be superior in revealing early, pre-visual symptoms and to better discriminate between the lines with different susceptibility to Cercospora.     

几种针阔叶树种不同光照强度下叶绿素荧光特性的研究

测定研究了油松、新疆杨等7种针阔叶树种不同光照强度下最大量子效率(ΦP)和实际量子效率(PhiPSⅡ)等叶绿素荧光特性的变化规律,进而研究了树木对光照强度适应性。结果表明,ΦP和PhiPSⅡ随光强度增加呈下降趋势,二者的光进程变化呈正相关直线。运用方程获得的量子效率最大点、光进程变化转折点和量子效率最小点,可评价树木光合量子效率的大小和对强光的适应性。依PhiPSⅡ最大值对被试树种光合量子效率排序为沙棘>白榆>新疆杨>白腊>油松>白木千>侧柏;依ΦP光进程变化转折点和量子效率最小点对应的光强度评价被试树木对强光的适应能力排序为新疆杨>白榆>侧柏>白腊>油松>沙棘>白木千,结果与被试树种生态特征基本一致。     

Age-dependent response of maize leaf segments to cadmium treatment: effect on chlorophyll fluorescence and phytochelatin accumulation

The relationship between the age of leaf tissue and response of the photosynthetic apparatus and phytochelatin accumulation to Cd treatment was studied. Studies were carried out with seedlings of Zea mays L. cv. Hidosil grown in the presence of 100-200 mumol/L Cd for 14 days under low light conditions. The third leaf was divided into 3 segments of equal length differing in the stage of tissue maturity and used for measurements of chlorophyll content, chlorophyll fluorescence, glutathione and phytochelatin content and Cd accumulation. A close relationship between the age of leaf tissue and response of the photosynthetic apparatus to Cd was shown. Cadmium (200 mumol/L) reduced photochemical processes more in older than younger leaf segments as seen in the Chl fluorescence parameters Fv/F0, and t1/2, while the chlorophyll fluorescence decrease ratio (Rfd) was inhibited more strongly in younger ones. Fv/Fm was slightly affected. Cd-induced enhancement of GSH content was correlated with higher phytochelatin accumulation to a greater extent in younger than in older leaf segments. Phytochelatin level corresponded to changes of photochemical processes in older leaves. The peptide thiol:Cd molar ratio for the phytochelatins varied depending on Cd concentration and age of leaf segments. The protective role of phytochelatins for the photosynthetic apparatus is discussed.     

Chlorophyll fluorescence and leaf chlorophyll content of bean leaves injured by spider mites (Acari: Tetranychidae)

The use of chlorophyll fluorescence as a method for detecting and monitoring plant stress arising from Tetranychus urticae (Koch) feeding injury was investigated. The effect of mite density (1–32 mites per 1.5 cm² of leaf) and the duration of the feeding period (1–5 days) on the chlorophyll fluorescence parameters of bean (Phaseolus vulgaris) leaves were examined. Changes in chlorophyll fluorescence parameters were dependent both on mite density and duration of feeding. Decreases in F _o, the initial fluorescence and F _m, the maximum fluorescence led to a decrease in the ratio of variable to maximum fluorescence, F _v/F _m. The decrease in F _v/F _m is typical of the response of many plants to a wide range of environmental stresses and indicates a reduced efficiency of photosystem II (PSII) photochemistry. T _1/2, which is proportional to the pool size of electron acceptors on the reducing side of PSII, was also reduced in response to mite-feeding injury. The leaf chlorophyll content decreased with increasing mite density and duration of feeding but did not appear to contribute to the decrease in F _v/F _m. Chlorophyll fluorescence is an effective method for detecting and monitoring stress in T. urticae-injured bean leaves.     

Chlorophyll distribution in the stems and trunk of beech trees

The distribution of chlorophyll was examined in cross-sections of 2- and 6-year-old stems as well as in the bark of the stump trunk of beech trees, utilising chlorophyll autofluorescence. The investigations were conducted using a confocal microscope. The tests carried out on 2 – 6-year old stems showed a large presence of chlorophyll in the bark, in primary and secondary rays as well as in the pith, but smaller amounts in wood parenchyma cells. There was no chlorophyll in the cork, sclerenchyma: in wood in vessels, tracheids and fibers. A reduction in the chlorophyll content in 6-year-old stems was not observed. In the bark of the trunk, chlorophyll was found in large amounts directly under the cork and in vestigial amounts in the primary phloem.     

氮素水平对不同品种茶树光合及叶绿素荧光特性的影响

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

干旱和复水对大豆(Glycine max)叶片光合及叶绿素荧光的影响

选用河南省大面积种植的大豆品种豫豆29作为实验材料,通过研究逐步干旱和旱后复水条件下大豆叶片光合、叶绿素荧光等指标随土壤水分的动态变化规律,以期为大豆的水分高效利用提供理论依据。研究发现,在土壤相对含水量高于46.5%时,虽然随着土壤相对含水量的下降,豫豆29仍可以保持它的叶片水分状态;豫豆29的叶片净光合速率在土壤水分中等条件下最大,在土壤相对含水量为64.3%时,它比对照组高出11.2%(P<0.01);在实验的第3d,处理组的土壤相对含水量降为46.5%,叶片水势与对照组相比降低了7.2%(P>0.05),净光合速率为对照组的89.6%(P<0.05),但气孔导度却迅速下降为对照组的44.7%(P<0.01),这说明与叶片的光合和水分状况相比,豫豆29的气孔对土壤水分的匮缺更加敏感。复水后,豫豆29叶片的水势、净光合速率、气孔导度和叶绿素荧光等值都可以得到迅速的恢复,并在实验的最后接近对照组的水平,这表明豫豆29的叶片光合在水分胁迫解除后有迅速恢复的能力。