Fluorescence induction in Avena sativa investigated by modulated light

The structurated slow fluorescence induction of six days old seedlings of Avena sativa L., cv. Seger I (Svalöf, Sweden) has been investigated in its responses to systematic changes in the exciting conditions. The intensity of the exciting light was periodically altered between a high and a low value, and after the third period the resulting fluorescence induction curves became stationary. These stationary responses may all be resolved into four characteristic fluorescence transients, whose forms were rather independent of the exciting conditions. The heights of these transients were examined as a function of the exciting wavelength and the high level intensity. From these data it is concluded that the third transient originates from cyclic electron transport and that the fourth transient monitors the degree of energy transfer from the light harvesting complex to photosystem II. These two last transients of the induction curve are equivalent to the structurated slow fluorescence changes in ordinary induction curves.     

Fluorescence induction kinetics of green and etiolated leaves by recording the complete in-vivo emission spectra

Complete room temperature fluorescence emission spectra of green and etiolated leaves (Raphanus sativus L., cv. Saxa Treib, Hordeum vulgare L., cv. Villa) are continuously recorded up to 4 min after onset of excitation. In green leaves two emission bands appear, whereas in etiolated leaves only one band is observed. In both cases the emission intensity increases with time, the high-energy band of green leaves decreasing more rapidly than the low-energy band. This phenomenon can be interpreted in terms of energy transfer. During the observation time of the fluorescence induction kinetic no shift of the emission peaks is found within the accuracy of the apparatus (±2nm).     

Kinetics of delayed chlorophyll a fluorescence registered in milliseconds time range

Delayed fluorescence dark decays in the time interval from 0.35 to 5.5ms are measured during dark to light adaptation in whole barley leaves and isolated thylakoid membranes, using a disc phosphoroscope. The changes in delayed fluorescence features are compared with variable chlorophyll fluorescence simultaneously registered with the same apparatus as well as in parallel by Handy PEA (Hansatech Instruments Ltd.), and absorbance changes at 820 nm. The registered delayed fluorescence signal is a sum of three components – submillisecond with lifetime of about 0.6 ms, millisecond decayed 2–4 ms and slow component with lifetime > >5.5 ms. The submillisecond delayed fluorescence component is proposed to be a result of radiative charge recombination in Photosystem II reaction centers in the state Z⁺PQ_A⁻Q_B⁻, and its lifetime is determined by the rate of electron transfer from Q_A⁻ to Q_B⁻. The millisecond delayed fluorescence component is associated with recombination in Z⁺PQ_A⁻Q_B⁼ centers with a lifetime determined by the sum of the rate constants of electron transfer from the oxygen-evolving complex to Z⁺ and of the exchange between the reduced and oxidized plastoquinone pool in the Q_B-site. On the basis of these assumptions and of the different share of the three components in the integral delayed fluorescence during induction, an attempt has been made to interpret the changes in the delayed fluorescence intensity during the transition of the photosynthetic apparatus from dark to light adapted state.     

超级高产小麦旗叶荧光动力学研究

探明超级小麦品种的旗叶光合作用与荧光动力学特性,为超级小麦品种选育利用提供理论依据。以超级小麦临麦4号为试验材料,应用CI-301PS型便携式光合作用测定系统和FMS-2便携式荧光测定仪(英国Hansatech公司)在田间试验中测定旗叶光合作用与荧光动力学参数。结果表明,与普通高产对照品种皖麦52和烟农19相比,超级小麦临麦4号的光合作用参数光合速率、光饱和点和CO2饱和点、羧化效率高,光补偿点和CO2补偿点低;光合机构系统工作参数PSII实际的光化学效率(ΦPSII)、光化学猝灭系数(qP)、PSII反应中心的激发能捕获效率(Fv/Fm)、PSⅡ潜在活性Fv/Fo和电子传递速率(ETR)值高,非光化学猝灭系数(NPQ)值低。这表明超级小麦临麦4号的光合机构系统工作能力强和工作效率高,保证旗叶光合作用的高效运行,为子粒灌浆提供充足的能量和碳水化合物。     

Rapid light-response curves of chlorophyll fluorescence in microalgae: relationship to steady-state light curves and non-photochemical quenching in benthic diatom-dominated assemblages

Rapid light-response curves (RLC) of variable chlorophyll fluorescence were measured on estuarine benthic microalgae with the purpose of characterising its response to changes in ambient light, and of investigating the relationship to steady-state light-response curves (LC). The response of RLCs to changes in ambient light (E, defined as the irradiance level to which a sample is acclimated to prior to the start of the RLC) was characterised by constructing light-response curves for the RLC parameters α _RLC, the initial slope, ETR_m,RLC, the maximum relative electron transport rate, and E _k,RLC, the light-saturation parameter. Measurements were carried out on diatom-dominated suspensions of benthic microalgae and RLC and LC parameters were compared for a wide range of ambient light conditions, time of day, season and sample taxonomic composition. The photoresponse of RLC parameters was typically bi-phasic, consisting of an initial increase of all parameters under low ambient light (E < 21–181 μmol m⁻² s⁻¹), and of a phase during which α _RLC decreased significantly with E, and the increase of ETR_m,RLC and E _k,RLC was attenuated. The relationship between RLC and LC parameters was dependent on ambient irradiance, with significant correlations being found between α _RLC and α, and between ETR_m,RLC and ETR_m, for samples acclimated to low and to high ambient irradiances, respectively. The decline of α _RLC under high light (Δα _RLC) was strongly correlated (P < 0.001 in all cases) with the level of non-photochemical quenching (NPQ) measured before each RLC. These results indicate the possibility of using RLCs to characterise the steady-state photoacclimation status of a sample, by estimating the LC parameter E _k, and to trace short-term changes in NPQ levels without dark incubation.     

Relationship between banding and photosynthetic activity in Chara corallina as studied by the spatially different induction curves of chlorophyll fluorescence observed by an image analysis system

Image analysis was applied for studying the kinetics of chlorophyll fluorescence in the alkaline and acid zones of internodal cells of Chara corallina Klein ex Willd., in order to test the hypothesis that in the acid bands an outward proton current yields a conversion of HCO₃⁻ ions to CO₂, thus facilitating inward diffusion of CO₂.
The analysis of the responses of chlorophyll fluorescence induced by changes in light intensity from 0.15 W m⁻² to 2.0 W m⁻² (λ <580 nm) revealed different kinetic behaviour. In the alkaline zones, the amplitude of the variable components of fluorescence was smaller than in the acid zones. Curve fitting by a sum of exponentials showed that the 4 time constants of about 2, 22, 48 and 140 s were not significantly different between the alkaline and acid zones. In contrast the related amplitude factors were smaller in the alkaline zones. by a factor of about 1.6 to 2.0 in old calcified cells and by a factor of about 1.2 in young cells without acrustations.
The interpretation of these results in terms of a better supply of carbon dioxide in the acid zones was supported by investigations on leaves of tobacco. Lowering the CO₂ concentration of an artificial atmosphere below 20 ppm resulted in a similar reduction of the variable component of chlorophyll fluorescence as found in the alkaline zones of Chara .     

不同发育时期冬小麦旗叶的荧光特性研究

首次对4种不同品种冬小麦（Triticum aestivumL.)的旗叶的诱导荧光动力学参数和最大净光合速率（Psat)进行了不同时期的比较,随着小麦从所花期到乳熟期的生长,旗叶的光系统II最大光量子效率（Fv/Fm)变化不大,在乳熟期略有下降,光化学淬灭（qP),光系统II量子产率（φPSII与）和Psat有较大的降低（＞15％）,非光化光学淬灭（qN)均有明显的增大（＞100％）,旗叶的φPSII与Psat的存线性关系（r=0.918),说明了在不同小麦品种中生长的衰老使得旗叶光合作用从能量转化到二氧化碳同化速率都降低。     

Technique of the Modulated Chlorophyll Fluorescence: Basic Concepts,Useful Parameters,and Some Applications

The review summarizes basic information about slow and fast chlorophyll (Chl) a fluorescence induction kinetics (FIK) recorded using fluorimeters working on a principle of the pulse amplitude modulation (PAM) of a Chl fluorescence signal. It explains fundamental principles of the measuring technique, evaluates the terminology, symbols, and parameters used. Analysis of Chl FIK resulting in a set of Chl fluorescence parameters (FPs) provides qualitative and quantitative information about photosynthetic processes in chloroplasts. Using FPs, one can describe the functioning of the photosynthetic apparatus under different internal and external conditions. Brief comments on proper application of the fluorimetric method in photosynthesis research and some actual examples are also given. This revised version was published online in June 2006 with corrections to the Cover Date.     

基于电子回流理论的植物延迟荧光来源理论与实验研究

延迟荧光(DF)技术在植物学、农业和环境等领域有着广泛的应用前景,延迟荧光来源的机理研究对其应用起着重要的作用。基于广泛接受的电子回流理论由一系列的光化学反应方程推导得出多指数衰减的延迟荧光动力学模型,首次提出了一个简化的三指数衰减近似模型,描述了光合电子传递链上的三个最重要成分的电子回流。为了验证近似模型的准确性,以实验室培养的相似的8周大的Jin Dan No.39玉米叶片为模型样品,利用实验室自制的延迟荧光探测系统,得到延迟荧光衰减动力学曲线。分别用三种电子阻断剂:DCMU、DB-MIB和Paraquat处理后的衰减动力学的实验与理论模型进行了对比研究,通过曲线特征来确定不同成分的存在及依赖关系。结果表明延迟荧光衰减动力学三指数拟合非常好(R2=0.99909),而且这三种成分是相互独立的,分别来源于QA,QB和光系统Ⅰ环式电子流。简化模型的提出为延迟荧光的应用题提供一个平台。     

Studies on the Characteristics of Chlorophyll Fluorescence of Winter Wheat Flag Leaves at Different Developing Stages

The parameters of fluorescence induction kinetics and the maximal light-saturated net CO2 assimilation rate ( P sat ) of the flag leaves of four cultivars of winter wheat （Triticum aestivum L.） were compared at three different developing stages for the first time. From the blooming stage to the milky stage, the quantum efficiency of PSⅡ photochemistry ( Fv/Fm ) declined slightly only at the milk stage. The photochemical quenching co-efficient ( qP ), actual quantum yield of photosystem Ⅱ（PSⅡ）electron transport ( Φ PSⅡ) and P sat decreased substantially (>15%), while the non-photochemical quenching co-efficient ( qN ) increased significantly (>100%). There existed a linear correlation between the Φ PSⅡ and the P sat ( r =0.918). The results indicate that with the senescence of the flag leaves of winter wheat the photosynthetic efficiency including that of the energy transport and the CO2 assimilation significantly decreased.     

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.     

Fluorescence induction of Photosystem II membranes shows the steps till reduction and protonation of the quinone pool

Chlorophyll fluorescence induction (Chl-F) was investigated in Photosystem II (PSII)-enriched membranes, which predominantly include active (Q_B reducing) PSII reaction centres (RCs) and lack Photosystem I (PSI). The Chl-F curve of these preparations show a polyphasic rise from F₀, the minimal fluorescence, to F_P, the maximal fluorescence, with several intermediate transitions. Analyses of these transitions revealed three exponential rise components with lifetimes of 18 ms, 400 ms and 800 ms. The 18 ms component was assigned to the photoaccumulation of reduced Q_A. The two slowest components, of 400 ms and 800 ms, were assigned to Q_B reduction (Q_B⁻ and Q_B⁼) and further Q_B⁼ protonation (till Q_BH₂), respectively. These assignments were based on the observation of specific quenching of the phases by DCMU or by different oxidized, reduced and protonated quinones. The work is done in low light conditions which are saturating to avoid photoinhibition or PSII inactivation effects. The results suggest that the Chl-F curve observed in PSII-enriched membranes can be attributed to the sequential steps till the photoaccumulation (reduction and protonation) of plastoquinone (PQ) by PSII. These results are in good agreement with the molecular models that show a correspondence between Chl-F and PQ reduction steps, like the models that propose and explain the O-J-I-P transients.     

Fluorescence induction and activity of ferredoxin-NADP reductase in Bryopsis chloroplasts

Effects of medium osmolarity on the rate of CO₂ fixation, the rate of the NADP⁺-Hill reaction, and the DPS₁ transient of chlorophyll fluorescence were measured in intact Bryopsis chloroplasts. Upon decreasing the sorbitol concentration from 1.0 M (the isoosmotic conditions) to 0.25 M, the envelopes of the chloroplasts became leaky to small molecules, resulting in a considerable depression of the CO₂-fixation rate and a higher rate of the NADP⁺-Hill reaction whereas the DPS₁ transient was unaffected. This DPS₁ transient of chlorophyll fluorescence is thought to be caused by the photoactivation of electron flow on the reducing side of Photosystem I at a site occurring after ferredoxin and probably before the reduction of NADP⁺ (Satoh, K. and Katoh, S. (1980) Plant and Cell Physiol. 21, 907–916). Little effect of NADP⁺ on the DPS₁ transient and a marked lag in NADP⁺ photo-reduction in dark-adapted (inactivated) chloroplasts support the hypothesis that the site of dark inactivation is prior to the reduction site of NADP⁺, and therefore, that ferredoxin-NADP⁺ reductase is inactivated in the dark and activated in the light. Moreover, at 0.25 M sorbitol, the activity of ferredoxin-NADP⁺ reductase itself (2,6-dichlorophenolindophenol reduction by NADPH) was shown to increase according to dark-light transition of the chloroplasts. At low osmolarities (below 0.1 M sorbitol), the difference in the diaphorase activity between dark-and light-adapted chloroplasts and the lag time observed in the NADP⁺ photoreduction were lowered. This may correspond to a less pronounced DPS₁ transient at low concentrations of sorbitol. The mechanism of the photo-activation is discussed.     

On the lag phase and initial decline of microbial growth curves

The lag phase is generally thought to be a period during which the cells adjust to a new environment before the onset of exponential growth. Characterizing the lag phase in microbial growth curves has importance in food sciences, environmental sciences, bioremediation and in understanding basic cellular processes. The goal of this work is to extend the analysis of cell growth curves and to better estimate the duration of the lag phase. A non-autonomous model is presented that includes actively duplicating cells and two subclasses of non-duplicating cells. The growth curves depend on the growth and death rate of these three subpopulations and on the initial proportion of each. A deterministic and a stochastic model are both developed and give the same results. A notable feature of the model is the decline of cells during the early stage of the growth curve, and the range of parameters when this decline occurs is identified. A limited growth model is also presented that accounts for the lag, exponential growth and stationary phase of microbial growth curves.     

Fluorescence from abnormally sterile pollen of the Japanese apricot

We observed trees of the Japanese apricot, Prunus mume ‘Nanko’ (Rosaceae), bearing two types of flowers: 34% had blue fluorescent pollen under UV irradiation, and 66% had non-fluorescent pollen. The fluorescent pollen grains were abnormally crushed, sterile, and devoid of intine and pollenkitt. The development of microspores within anthers was investigated: in the abnormally developed anthers, tapetal cells were vacuolated at the unicellular microspore stage, and fluorescent pollen was produced. Compounds responsible for the blue fluorescence of pollen were identified as chlorogenic acid and 1-O-feruloyl-β-D-glucose. The anthers with fluorescent pollen contained 6.7-fold higher and 3.8-fold lower amounts of chlorogenic acid and N¹,N⁵,N¹⁰-tri-p-coumaroylspermidine, respectively, compared to those with non-fluorescent pollen. The tapetal vacuolization, highly accumulated chlorogenic acid, and deficiency of N¹,N⁵,N¹⁰-tri-p-coumaroylspermidine imply that low-temperature stress during the early unicellular microspore stage caused a failure in microsporogenesis. Furthermore, potential effects of the visual difference on the bee behavior were also discussed through the colorimetry. The sterility, likely induced by low-temperature stress, and the preference of honeybees for fluorescence may reduce the pollination efficiency of P. mume.     

沙地生境沙拐枣(Calligonum mongolicum)种群特征及其扩张

沙拐枣(Calligonum mongolicum)是分布于我国北方荒漠、半荒漠的一种主要固沙植物,研究沙拐枣种群在沙地生境中的扩张对沙漠-绿洲过渡带的生态恢复和重建管理具有重要的实践意义.以巴丹吉林沙漠延伸带与河西走廊中部张临高绿洲过渡带为研究区域,采用踏查与样方调查相结合的方法对4种不同沙地生境(固定沙丘、半流动沙丘迎风坡、背风坡、丘间低地)中沙拐枣种群特征、繁殖规律、种群的扩张域和扩张速率进行了研究.结果表明:(1) 沙地生境中沙拐枣以有性繁殖为主.固定沙丘、半流动沙丘丘间低地、迎风坡和背风坡萌蘖苗密度占总密度的比值分别为35%、45%、2%、9%;实生苗密度比值分别为65%、55%、98%、91%.(2)不同生境中沙拐枣种群都为增长型种群;(3) 固定沙丘、半流动沙丘迎风坡、背风坡、丘间低地中,沙拐枣种群无性繁殖的扩张域和平均扩张速率分别为1～5m和0.67m/a、1～8m和0.64m/a、0～2m和0.15m/a、1～4m和0.41m/a.同时,3种生境中沙拐枣种群密度及其扩张速率都随着年龄的增长而下降.(4)沙拐枣种群在侵入新的生境过程中,开始于有性繁殖,利用无性繁殖拓展生存空间.     

Integrating individual behavior and population ecology: the potential for habitat-dependent population regulation in a reef fish

We used the predictions of the ideal free and ideal despoticdistributions (IFD and IDD, respectively) as a basis to evaluatethe link between spatial heterogeneity, behavior, and populationdynamics in a Caribbean coral reef fish. Juvenile three-spotdamselfish (Stegastes planifrons) were more closely aggregatedin patch reef habitat than on continuous back reef. Agonisticinteractions were more frequent but feeding rates were lowerin the patch versus the continuous reef habitat. Growth rateswere lower in patch reef habitat than on the continuous reef,but mortality rates did not differ. A separate experiment usingstandard habitat units demonstrated that the patterns observedin natural habitat were the result of the spatial distributionof the habitat patches rather than resource differences between habitats. Our results do not follow the predictions of simpleIFD or IDD models. This deviation from IFD and IDD predictionsmay be the result of a number of factors, including lack ofperfect information about habitat patches, high movement costs,and higher encounter rates of dispersed patches. Our resultsdemonstrate that behavioral interactions are an integral partof population dynamics and that it is necessary to considerthe spatial organization of the habitat in both behavioraland ecological investigations.     

Transition from a random to an ideal free to an ideal despotic distribution: the effect of individual difference in growth

Individual differences in growth can lead to a monopolistic form of food competition. We studied the long-term transition in the mode of competition and the distribution of individuals between food patches of the cloned salmonid fish, Oncorhynchus masou ishikawae, in the laboratory. This transition was accompanied by growth depensation, i.e., the increase over time in the variance of size between individuals resulting from the differences in individual growth rates. The 120-cm experimental tanks were divided into two compartments (patches) between which an opaque partition was placed. Fish were able to move freely between the patches and therefore were able to assess the patch quality using long-term memory, but they were not able to see the food input in the other patch directly. The distribution between the two food patches, the amount of food gained, and the growth and the agonistic behavior of four groups of six individuals were observed over 4 weeks. We found that (1) within-group variation in body weight increased with time; (2) on average, the better patch was used by more individuals than predicted by a random distribution but fewer individuals than predicted by an ideal free distribution, and (3) the distribution and pattern of resource use by the fish changed over the 4-week experimental period from a random distribution to an ideal free distribution and finally to an ideal despotic distribution. We suggest that growth depensation causes the long-term change in the spatial distribution and pattern of resource use by competitors. Received: December 19, 2000 / Accepted: March 19, 2001     

Secondary follicle growth and oocyte maturation by culture in alginate hydrogel following cryopreservation of the ovary or individual follicles

An option for fertility preservation for women facing a cancer diagnosis involves the cryopreservation of ovarian tissue for later re‐transplantation or in vitro culture, with in vitro culture preferred to avoid reintroduction of the cancer. Small, immature follicles survive the freeze‐thaw process, and can be matured through in follicle maturation (IFM) that involves an initial growth of the follicle and subsequent maturation of the oocyte. The ovarian tissue can be cryopreserved in two forms: (i) cortical strips consisting of follicles and surrounding stroma (Cryo‐Ov) or (ii) individually isolated follicles (Cryo‐In). The aim of this study was to assess the follicle growth and oocyte maturation for follicles that were cryopreserved either as strips or individually using a slow‐freezing cryopreservation method. The two follicle groups, together with non‐cryopreserved control follicles, were grown in an alginate‐based three‐dimensional culture system for 12 days. The overall survival, size increase and antrum formation rates were comparable among the three groups. At day 12 of culture, Androstenedione levels were decreased in the Cryo‐Ov group relative to the other two, and the ratio of progesterone to estradiol was increased in the two cryopreserved groups relative to the control. Both Gja1 (known as connexin 43) and Gja4 (known as connexin 37) mRNA expression were decreased at day 6 in the cryopreserved groups relative to controls, and by day 12, Gja1 was similar for all three groups. Moreover, Cryo‐In resulted in lower GVBD rate indicating some impaired oocyte development. Overall, the present study demonstrated that mouse preantral follicles, either within ovarian tissues or individually isolated, could be successfully cryopreserved by the slow‐freezing method, as evidenced by post‐thaw follicle development and steroidgenesis, oocyte maturation and molecular markers for oocyte and/or granulosa cells connection. Biotechnol. Bioeng. 2009;103: 378–386. © 2009 Wiley Periodicals, Inc.