首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 31 毫秒
1.
Hopt A  Neher E 《Biophysical journal》2001,80(4):2029-2036
Two-photon fluorescence excitation is being increasingly used in laser scan microscopy due to very low photodamage induced by this technique under normal operation. However, excitation intensity has to be kept low, because nonlinear photodamage sets in when laser power is increased above a certain threshold. We studied this kind of damage in bovine adrenal chromaffin cells, using two different indicators of damage: changes in resting [Ca(2+)] level and the degranulation reaction. In agreement with previous studies, we found that, for both criteria, damage is proportional to the integral (over space and time) of light intensity raised to a power approximately 2.5. Thus, widening the laser pulse shape at constant average intensity both in time and in focal volume is beneficial for avoiding this kind of damage. Both measures, of course, reduce the two-photon fluorescence excitation. However, loss of signal can be compensated by increasing excitation power, such that, at constant damaging potential, signals may be even larger with long pulses and large focal volumes, because the exponent of the power law of damage is higher (mu approximately 2.5) than that of the two-photon signal (mu approximately 2).  相似文献   

2.
Excitation kinetics based on feedback regulation of chlorophyll (Chl) fluorescence of leaves measured with the chlorophyll fluorometer, FluoroMeter Modul (FMM), are presented. These kinetics showed the variation of excitation light (laser power, LP) regulated by the feedback mechanism of the FMM, an intelligent Chl fluorometer with embedded computer, which maintains the fluorescence response constant during the 300-s transient between the dark- and lightadapted state of photosynthesis. The excitation kinetics exhibited a rise of LP with different time constants and fluctuations leading to a type of steady state. The variation of excitation kinetics were demonstrated using the example of primary leaves of etiolated barley seedlings (Hordeum vulgare L. cv. Barke) during 48 h of greening in the light with gradual accumulation of Chl and development of photosynthetic activity. The excitation kinetics showed a fast rise followed by a short plateau at ca. 30 s and finally a slow constant increase up to 300 s. Only in the case of 2 h of greening in the light, the curve reached a stable steady state after 75 s followed by a slight decline. The final LP value (at 300 s of illumination) increased up to 12 h of greening and decreased with longer greening times. The active feedback mechanism of the FMM adjusted the excitation light during the measurement to the actual photosynthetic capacity of the individual leaf sample. In this way, the illumination with excessive light was avoided. The novel excitation kinetics can be used to characterize health, stress, disease, and/or product quality of plant material.  相似文献   

3.
胡文海  肖宜安 《植物研究》2022,42(6):1052-1061
植物叶片光合作用具有高度的空间异质性,叶绿素荧光成像技术为叶片光合异质性的研究提供了便利,但叶片光合异质性的定量分析并没有得到广泛应用。本文利用Imaging PAM叶绿素荧光成像系统,获得中亚热带地区越冬期小叶榕(Ficus microcarpa)阳生叶和阴生叶的叶绿素荧光参数图像,并利用仪器的分析软件对其进行分析,定量比较了阳生叶和阴生叶的光合异质性特征。研究发现:越冬期小叶榕阳生叶的光合异质性和光抑制程度明显高于阴生叶,变异系数可作为光合异质性的定量指标。低温强光导致阳生叶坏死率(PLN)达4.30%,并有53.30%的区域处于严重光抑制(0v/Fm<0.627),但仍有42.27%的区域仅为轻度光抑制(0.627≤Fv/Fm<0.800)。而低温弱光并未造成阴生叶坏死和严重光抑制。通过对光系统Ⅱ(PSⅡ)的实际光合效率(Y(Ⅱ))、调节性能量耗散的量子产额(Y(NPQ))和非调节性能量耗散的量子产额(Y(NO))荧光参数异质性的定量分析表明,阳生叶...  相似文献   

4.
Abstract. The effect of chlorophyll fluoresence on the spectral light gradients within a model green leaf was examined under different light qualities (day-light, sunset, canopy) and different quantum efficiencies. Light fluxes within the leaf tissue are nearly doubled in the emission domain of fluorescence but the effect on the phytochrome photoequilibrium is very small.  相似文献   

5.
A midday depression in net photosynthesis and in stomatal conductancewas observed when leaves of well-watered Vitis vinifera plantswere subjected to a diurnal pattern of variation in leaf temperatureand leaf-to-air water vapour pressure difference similar toa summer day, while photon flux density was kept constant at1450 µmolm–2 s–1,. When leaves were kept atconstant leaf temperature (22.5°C) and leaf-to-air watervapour presure difference (8.5 Pa kPa–1) at the same lightintensity, stomata opened with the onset of illumination andmaximal conductance and photosynthesis values were observedabout 1 h later. Subsequently, conductance and photosynthesisdecreased gradually. Leaf water potential never dropped below{macron}0.3 MPa. Leaves kept under constant environmental conditionsshowed an afternoon decline in photosynthesis at high internalCO2, in carboxylation efficiency and in maximum conductanceas well as an increase in stomatal sensitivity to CO2. Whenthe photon flux density during the day was reduced to 750 µmolm–2 s–1, the afternoon depression in gas exchangerates was attenuated. To evaluate the possible effects of highlight stress on changes in chloroplastic behaviour we comparedlight response curves of photosynthesis determined with an oxygenelectrode, in the morning and in the afternoon after the plantswere exposed to either high or moderate photon flux densities.A significant depression in photosynthetic capacity was foundby this method in high light treated leaves, but not in leavespreviously exposed to moderate photon flux density. Apparentquantum yield decreased in the afternoon, particularly afterexposure to high light. Maximum chlorophyll a fluorescence at22°C was reduced and the quenching of fluorescence afterreaching the peak was slower in the afternoon than in the morning,especially in high light-treated leaves. Changes in the patternsof chlorophyll fluorescence kinetics were observed after lighttreatment, i.e. in the afternoon, with oscillations either absent(after high light) or significantly reduced (after moderatelight) in comparison to the morning. The significance of theseresults is discussed and it is suggested that a direct inhibitoryeffect of high light at the chloroplast level provides the bestinterpretation for the observed afternoon decline in photosyntheticrate. Key words: Carboxylation efficiency, chlorophyll fluorescence, photosynthetic capacity, quantum yield, stomatal conductance, Vitis vinifera L.  相似文献   

6.
Y. Kobayashi  S. Köster  U. Heber 《BBA》1982,682(1):44-54
Scattering of green light and chlorophyll fluorescence by spinach leaves kept in a stream of air or nitrogen were compared with leaf adenylate levels during illumination with blue, red or far-red light. Energy charge and ATP-ADP ratios exhibited considerable variability in different leaves both in the dark and in the light. Variability is explained by different possible states of the reaction oxidizing triose phosphate or reducing 3-phosphoglycerate. Except when oxygen levels were low, there was an inverse relationship between light scattering and chlorophyll fluorescence during illumination with blue or red light. When CO2 was added to a stream of CO2-free air, chlorophyll fluorescence increased, sometimes after a transient decrease, and both light scattering and leaf ATPADP ratios decreased. Similar observations were made when air was replaced by nitrogen under blue or high-intensity red light. Under these conditions, over-reduction caused inhibition of electron transport and phosphorylation in chloroplasts. However, when air was replaced by nitrogen during illumination with low-intensity red light or far-red light, light scattering increased instead of decreasing. Under these light conditions, ATPADP ratios were maintained in the light. They decreased drastically only after darkening. Although ATPADP ratios responded faster than light scattering or the slow secondary decline of chlorophyll fluorescence due to illumination, it appeared that in the steady state, light scattering and chlorophyll fluorescence are useful indicators of the phosphorylation state of the leaf adenylate system at least under aerobic conditions, when chloroplast and extrachloroplast adenylate systems can effectively communicate.  相似文献   

7.
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 (φabs) 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.  相似文献   

8.
9.
The UV light (337 nm) induced blue-green fluorescence emission of green leaves is characterized at room temperature (298 K) by a maximum near 450 nm (blue region) and a shoulder near 525 nm (green region) and was here also studied at 77 K. At liquid nitrogen temperature (77 K) the blue (F450) and green fluorescence (F525) are much enhanced as is the red chlorophyll fluorescence near 735 nm. During development of green tobacco leaves the blue fluorescence F450 (77 K) is shifted towards longer wavelengths from about 410 nm to 450 nm. The isolated leaf epidermis of tobacco showed only slight fluorescence emission with a maximum near 410 nm. The green fluorescence F525 was found to mainly originate from the mesophyll of the leaf, its intensity increased when the epidermis was removed. The red chlorophyll fluorescence emission was also enhanced when the epidermis was stripped off; this considerably changed the blue/red fluorescence ratios F450/F690 and F450/F735. The epidermis, with its cell wall and UV-light-absorbing substances in its vacuole, plays the role of a barrier for the exciting UV-light. In contrast to intact and homogenized leaves, isolated intact chloroplasts and thylakoid membranes did not exhibit a blue-green fluorescence emission.  相似文献   

10.
Chlorophyll fluorescence was used to estimate profiles of absorbed light within chlorophyll solutions and leaves. For chlorophyll solutions, the intensity of the emitted fluorescence declined in a log–linear manner with the distance from the irradiated surface as predicted by Beer's law. The amount of fluorescence was proportional to chlorophyll concentration for chlorophyll solutions given epi‐illumination on a microscope slide. These relationships appeared to hold for more optically complex spinach leaves. The profile of chlorophyll fluorescence emitted by leaf cross sections given epi‐illumination corresponded to chlorophyll content measured in extracts of leaf paradermal sections. Thus epifluorescence was used to estimate relative chlorophyll content through leaf tissues. Fluorescence profiles across leaves depended on wavelength and orientation, reaching a peak at 50–70 µm depth. By infiltrating leaves with water, the pathlengthening due to scattering at the airspace : cell wall interfaces was calculated. Surprisingly, the palisade and spongy mesophyll had similar values for pathlengthening with the value being greatest for green light (550 > 650 > 450 nm). By combining fluorescence profiles with chlorophyll distribution across the leaf, the profile of the apparent extinction coefficient was calculated. The light profiles within spinach leaves could be well approximated by an apparent extinction coefficient and the Beer–Lambert/Bouguer laws. Light was absorbed at greater depths than predicted from fibre optic measurements, with 50% of blue and green light reaching 125 and 240 µm deep, respectively.  相似文献   

11.
Ball MC  Critchley C 《Plant physiology》1982,70(4):1101-1106
Photosynthetic responses to irradiance by the grey mangrove, Avicennia marina (Forstk.) Vierh var. australasica (Walp.) Moldenke, were studied using seedlings grown under natural understory shade and exposed conditions as well as in the laboratory under high and low light regimes, i.e. 100% and 6% sunlight, respectively. Leaves in exposed locations were subjected to daylight quantum flux densities greater than 1,000 microeinsteins per square meter per second from 0900 to 1700 hours, whereas those in understory shade experienced only 30 to 120 microeinsteins per square meter per second, interrupted for brief periods by sunflecks ranging in quantum flux density from 800 to 1,500 microeinsteins per square meter per second. The low light regime was similar in light intensity to that of the understory environment, but lacked sunflecks.

Leaves from the understory environment showed several properties of `shade' leaves; i.e. they contained more chlorophyll on both a leaf area and fresh weight basis but had a lower specific weight and greater area than exposed leaves, and were enriched in chlorophyll b relative to a. However, there were no significant differences in either the gas exchange or leaf chlorophyll fluorescence characteristics of the two populations, both being typical of `sun' leaves.

Leaves grown in the laboratory under low and high light regimes had similar properties. However, light saturated assimilation rates in the leaves from the low light treatment were 20% less and became light saturated at a lower quantum flux density than those of leaves grown under the high light regime. The ecological significance of these results is discussed.

  相似文献   

12.
The use of fluorescence induction measurements in leaves infiltrated with 3-(3,4-dichlorophenyl)-1,1-dimethylurea has been evaluated as a routine method for estimation of the concentration of the reaction centers of photosystem II relative to total chlorophyll in a wide variety of plant species. The procedure is based on a simple theory that takes into account the attenuation of light in passing through the leaf and the linear dependence of the fluorescence induction time from different parts of the leaf on the inverse of the local light intensity. A formula to calculate the reaction center concentration of photosystem II was obtained. The effect of the light attenuation is accounted for by a correction factor which could become practically insignificant by an optimal choice of the excitation and emission wavelengths and the geometry of the photodetector with respect to the sample. Estimation of quantum yields for primary photochemistry and influence of light scattering were considered. The results demonstrate the effect of the above factors under various circumstances and are in agreement, to a first approximation, with the theory.  相似文献   

13.
比较研究了‘早美’和‘春蕾’2个早熟桃品种夏季叶色转红对太阳光能的利用和光系统Ⅱ的叶绿素荧光特征的影响。结果表明:早熟桃叶片色素组成的变化会显著影响其光合和叶绿素荧光特性。叶色转红后,早熟桃净光合速率(Pn)日均值、PSII最大光化学效率(Fv/Fm)、PSII实际光化学效率(ФPSII)均上升,无显著光抑制,而绿叶对照‘红花碧桃’的电子传递速率(ETR)、Fv/Fm和ФPSII值均显著下降,7月光合明显受抑制。叶色转红程度较深的‘早美’在夏季高温强光下表现优于‘春蕾’和对照。淬灭分析表明:叶片花色素苷的积累能在短时间内增加PSII天线色素吸收的光能用于光化学反应的份额(P)与用于反应中心热耗散的相对份额(D)。转红后的叶片光化学淬灭系数(qp)显著高于绿叶,PSII光化学效率较高,但耗散过剩激发能的能力显著低于绿叶对照。  相似文献   

14.
Leaves of the C3 plant Brassica oleracea were illuminated with red and/or far-red light of different photon flux densities, with or without additional short pulses of high intensity red light, in air or in an atmosphere containing reduced levels of CO2 and/or oxygen. In the absence of CO2, far-red light increased light scattering, an indicator of the transthylakoid proton gradient, more than red light, although the red and far-red beams were balanced so as to excite Photosystem II to a comparable extent. On red background light, far-red supported a transthylakoid electrical field as indicated by the electrochromic P515 signal. Reducing the oxygen content of the gas phase increased far-red induced light scattering and caused a secondary decrease in the small light scattering signal induced by red light. CO2 inhibited the light-induced scattering responses irrespective of the mode of excitation. Short pulses of high intensity red light given to a background to red and/or far-red light induced appreciable additional light scattering after the flashes only, when CO2 levels were decreased to or below the CO2 compensation point, and when far-red background light was present. While pulse-induced light scattering increased, non-photochemical fluorescence quenching increased and F0 fluorescence decreased indicating increased radiationless dissipation of excitation energy even when the quinone acceptor QA in the reaction center of Photosystem II was largely oxidized. The observations indicate that in the presence of proper redox poising of the chloroplast electron transport chain cyclic electron transport supports a transthylakoid proton gradient which is capable of controlling Photosystem II activity. The data are discussed in relation to protection of the photosynthetic apparatus against photoinactivation.Abbreviations F, FM, F'M, F"M, F0, F'0 chlorophyll fluorescence levels - exc quantum efficiency of excitation energy capture by open Photosystem II - PS II quantum efficiency of electron flow through Photosystem II - P515 field indicating rapid absorbance change peaking at 522 nm - P700 primary donor of Photosystem I - QA primary quinone acceptor in Photosystem II - QN non-photochemical fluorescence quenching - Qq photochemical quenching of chlorophyll fluorescence  相似文献   

15.
Profiles of chlorophyll fluorescence were measured in spinach leaves irradiated with monochromatic light. The characteristics of the profiles within the mesophyll were determined by the optical properties of the leaf tissue and the spectral quality of the actinic light. When leaves were infiltrated with 10?4M DCMU [3‐(3,4‐dichlorophenyl)‐1, 1‐dimethyl‐urea] or water, treatments that minimized light scattering, irradiation with 2000 μmol m?2 s?1 green light produced broad Gaussian‐shaped fluorescence profiles that spanned most of the mesophyll. Profiles for chlorophyll fluorescence in the red (680 ± 16 nm) and far red (λ > 710 nm) were similar except that there was elevated red fluorescence near the adaxial leaf surface relative to far red fluorescence. Fluorescence profiles were narrower in non‐infiltrated leaf samples where light scattering increased the light gradient. The fluorescence profile was broader when the leaf was irradiated on its adaxial versus abaxial surface due to the contrasting optical properties of the palisade and spongy mesophyll. Irradiation with blue, red and green monochromatic light produced profiles that peaked 50, 100 and 150 μm, respectively, beneath the irradiated surface. These results are consistent with previous measurements of the light gradient in spinach and they agree qualitatively with measurements of carbon fixation under monochromatic blue, red and green light. These results suggest that chlorophyll fluorescence profiles may be used to estimate the distribution of quanta that are absorbed within the leaf for photosynthesis.  相似文献   

16.
Blue (F 450) and green (F 530) leaf fluorescence were studied together with the red chlorophyll fluorescence (emission maxima F 690 and F 735) during light-induced chlorophyll fluorescence induction kinetics (Kautsky effect) in predarkened leaves of wheat (Triticum aestivum L.) and soybean (Glycine max L.). The intensity of the red chlorophyll fluorescence decreased from maximum fluorescence Fm to steady-state fluorescence Fs, and the fluorescence ratio F 690/F 735 decreased by about 10% from Fm to Fs. However, blue and green fluorescence intensities remained constant throughout the measuring time. Consequently, the ratio of blue to red fluorescence (F 450/F 690) increased during chlorophyll fluorescence induction kinetics, whereas the ratio of blue to green fluorescence (F 450/F 530) remained unchanged within the same period. The knowledge of these ratios will be a prerequisite for the interpretation of remote sensing data from terrestrial vegetation.  相似文献   

17.
以当年生盆栽金花茶实生苗为材料,研究不同程度的强光胁迫(25%、50%和100%自然光强,以8%自然光强为对照)对其生长、生物量、叶片光合色素含量、叶绿素荧光参数的影响。结果表明:在不同程度的强光胁迫下,金花茶幼苗的生长均受到抑制,随着胁迫程度的增强,金花茶叶片颜色由深绿变为浅绿、黄绿色,叶片灼伤愈来愈严重;植株抽稍时间推迟,抽稍后长出的新叶长势较差;幼苗死亡率越来越高。幼苗根生物量、茎生物量、叶生物量和总生物量均随胁迫程度的升高而显著降低,强光胁迫对叶生物量的影响最大,根生物量次之,对茎生物量的影响最小。随着胁迫程度的增强,叶片叶绿素总量(Chl)、叶绿素a(Chla)、叶绿素b(Chlb)含量均显著降低,Chla/Chlb和Car/Chl显著升高。叶绿素荧光参数FoFmFvFv/FmFv/Fo均随胁迫程度的升高降低,强光胁迫使PSⅡ受到了伤害,光合作用原初反应过程受抑制,光合电子传递受到影响,从而抑制植株的正常生长。  相似文献   

18.
Chlorophyll fluorescence constitutes a simple, rapid, and non-invasive means to assess light utilization in Photosystem II (PS II). This study examines aspects relating to the accuracy and applicability of fluorescence for measurement of PS II photochemical quantum yield in intact leaves. A known source of error is fluorescence emission at 730 nm that arises from Photosystem I (PS I). We measured this PS I offset using a dual channel detection system that allows measurement of fluorescence yield in the red (660 nm < F < 710 nm) or far red (F > 710 nm) region of the fluorescence emission spectrum. The magnitude of the PS I offset was equivalent to 30% and 48% of the dark level fluorescence F0 in the far red region for Helianthus annuus and Sorghum bicolor, respectively. The PS I offset was therefore subtracted from fluorescence yields measured in the far red spectral window prior to calculation of PS II quantum yield. Resulting values of PS II quantum yield were consistently higher than corresponding values based on emission in the red region. The basis for this discrepancy lies in the finite optical thickness of the leaf that leads to selective reabsorption by chlorophyll of red fluorescence emission originating in deeper cell layers. Consequently, red fluorescence measurements preferentially sense emission from chloroplasts in the uppermost layer of the leaf where levels of photoprotective nonphotochemical quenching are higher due to increased photon density. It is suggested that far red fluorescence, corrected for the PS I offset, provides the most reliable quantitative basis for calculation of PS II quantum yield because of reduced sensitivity of these measurements to gradients in leaf transmittance and quenching capacity. This revised version was published online in August 2006 with corrections to the Cover Date.  相似文献   

19.
1. The fluorescence spectra of the alga Porphyridium have been recorded as energy distribution curves for eleven different incident wave lengths of monochromatic incident light between wave lengths 405 and 546 mµ. 2. In these spectra chlorophyll fluorescence predominates when the incident light is in the blue part of the spectrum which is strongly absorbed by chlorophyll. 3. For blue-green and green light the spectrum excited in Porphyridium contains in addition to chlorophyll fluorescence, the fluorescence bands characteristic of phycoerythrin and of phycocyanin. 4. From these spectra the approximate curves for the fluorescence of the individual pigments phycoerythrin, phycocyanin, and chlorophyll in the living material have been derived and the relative intensity of each of them has been obtained for each of the eleven incident wave lengths. 5. The effectiveness spectrum for the excitation of the fluorescence of these three pigments in vivo has been plotted. 6. From comparisons of the effectiveness spectrum for the excitation of each of these pigments it appears that both phycocyanin and chlorophyll receive energy from light which is absorbed by phycoerythrin. 7. It is suggested that phycocyanin may be an intermediate in the resonance transfer of energy from phycoerythrin to chlorophyll. 8. Since phycoerythrin and phycocyanin transfer energy to chlorophyll, it appears probable that chlorophyll plays a specific chemical role in photosynthesis in addition to acting as a light absorber.  相似文献   

20.
环境强光诱导玉簪叶片光抑制的机制   总被引:2,自引:0,他引:2       下载免费PDF全文
为进一步阐述光抑制的强光诱导和发生机制, 该文以喜阴植物玉簪(Hosta spp.)为材料研究其光抑制发生规律及其与环境光强的关系。结果表明, 全日照和遮阴条件下玉簪叶片发育分别形成适应强光和弱光的形态特征; 与遮阴处理相比, 强光下生长的玉簪光合速率和叶绿素含量较低, 但两种处理叶片最大光化学效率差异很小, 证明强光下植株可以正常生长且光合机构未发生严重的光抑制。将遮阴处生长的植株转移到全日照下, 光合速率和最大光化学效率急剧下降; 荧光诱导动力学曲线发生明显改变, 而且光系统II供体侧和受体侧荧光产量的变化幅度分别达到24.3%和34.2%, 表明玉簪由弱光转入强光后光系统II发生不可逆失活, 且受体侧受到的伤害较供体侧更严重。因此, 作者认为环境光强骤然提高并超过玉簪生长光强时很容易诱导其光合机构发生严重的光抑制。该研究对于理解植物适应光环境的策略以及喜阴植物的优质栽培有重要意义。  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号