汽车尾气对四种北方阔叶树叶绿素荧光特性的影响

采用开顶式熏气室研究了汽车尾气对五角槭、山荆子、山梨和茶条槭 4树种叶绿素荧光参数的影响。采用了相同浓度不同处理时间和相同处理时间不同浓度两种方法进行熏气处理。结果表明 ,随着熏气时间的延长和熏气浓度的增加 ,4树种的荧光参数Fv/Fm、Fv/F0 、ΦPSII、qP均逐渐降低 ,qN则逐渐上升。 4树种荧光参数的变化幅度不同 ,其中山荆子变化幅度最大 ,茶条槭的变化幅度最小 ,五角槭和山梨介于两者之间。以叶绿素荧光参数评价 4树种抗污染胁迫能力 ,茶条槭最强 ,五角槭和山梨次之 ,山荆子最弱。     

Myoplasmic binding of fura-2 investigated by steady-state fluorescence and absorbance measurements.

Binding of the fluorescent Ca2+ indicator dye fura-2 by intracellular constituents has been investigated by steady-state optical measurements. Fura-2's (a) fluorescence intensity, (b) fluorescence emission anisotropy, (c) fluorescence emission spectrum, and (d) absorbance spectra were measured in glass capillary tubes containing solutions of purified myoplasmic proteins; properties b and c were also measured in frog skeletal muscle fibers microinjected with fura-2. The results indicate that more than half, and possibly as much as 85%, of fura-2 molecules in myoplasm are in a protein-bound form, and that the binding changes many properties of the dye. For example, in vitro characterization of the Ca2+-dye reaction indicates that when fura-2 is bound to aldolase (a large and abundant myoplasmic protein), the dissociation constant of the dye for Ca2+ is three- to fourfold larger than that measured in the absence of protein. The problems raised by intracellular binding of fura-2 to cytoplasmic proteins may well apply to cells other than skeletal muscle fibers.     

Effects of drought on photosynthesis,chlorophyll fluorescence and photoinhibition susceptibility in intact willow leaves

Plants from clonal cuttings of Salix sp. were subjected to a drying cycle of 10 d in a controlled environment. Gas exchange and fluorescence emission were measured on attached leaves. The light-saturated photosynthetic CO₂ uptake became progressively inhibited with decreased leaf water potential both at high, and especially, at low intercellular CO₂ pressure. The maximal quantum yield of CO₂ uptake was more resistant. The inhibition of light-saturated CO₂ uptake at leaf water potentials around-10 bar, measured at a natural ambient CO₂ concentration, was equally attributable to stomatal and non-stomatal factors, but the further inhibition below this water-stress level was caused solely by non-stomatal factors. The kinetics of fluorescence emission was changed at severe water stress; the slow secondary oscillations of the induction curve were attenuated, and this probably indicates perturbations in the carbon reduction cycle. The influence of light level during the drought period was also studied. Provided the leaves were properly light-acclimated, drought at high and low light levels produced essentially the same effects on photosynthesis. However, low-light-acclimated leaves became more susceptible to photoinhibitory treatment under severe water stress, as compared with well-watered conditions.     

Photoinhibition of photosystems I and II using chlorophyll fluorescence measurements

In this study the photoinhibition of photosystems (PS) I and II caused by exposure to high intensity light in oat (Avena sativa, var Prevision) is measured by the emission of chlorophyll fluorescence in intact leaves adapted to darkness. The maximal quantum yield of PS II was lower in plants grown under high light intensity than in plants grown under low intensity, which indicates that PS II is photoinhibited by such conditions. PS I was more stable than PS II in plants exposed to strong light for a moderate time (five photoperiods) since the oxidised plastoquinone pool size under far-red (FR) light was similar in plants grown under high light intensity to plants grown under low intensity, probably as a result of the cyclic electron flow around PS I being stimulated in response to high light intensity. However, over longer times (10 photoperiods) the PS I was photoinhibited, since the oxidised plastoquinone pool size under FR light increased as a consequence of the decrease in PS I activity caused by high light intensity. This practical is intended for advanced students of plant biochemistry and plant physiology.     

Estimating heat tolerance among plant species by two chlorophyll fluorescence parameters

The heat tolerance of 8 temperate- and 1 subtropical-origin C₃ species as well as 17 tropical-origin ones, including C₃, C₄, and CAM species, was estimated using both F₀-T curve and the ratio of chlorophyll fluorescence parameters, prior to and after high temperature treatment. When leaves were heated at the rate of ca. 1 °C min⁻¹ in darkness, the critical temperature (T_c) varied extensively among species. The T_c's of all 8 temperate-origin species ranged between 40–46 °C in winter (mean temperature 16–19 °C), and between 32–48 °C in summer (mean temperature ca. 30 °C). Those for 1 subtropical- and 12 tropical-origin C₃ species ranged between 25–44 °C and 35–48 °C, and for 1 CAM and 4 C₄ species were 41–47 and 45–46 °C, respectively. Acclimating three C₃ herbaceous plants at high temperature (33/28 °C, day/night) for 10 d in winter caused their T_c's rising to nearly the values measured in summer. When leaves were exposed to 45 °C for 20 min and then kept at room temperature in darkness for 1 h, a significant correlation between RF_v/m (the ratio of F_v/F_m before and after 45 °C treatment) and T_c was observed for all tested temperate-origin C₃ species as well as tropical-origin CAM and C₄ species. However, F₀ and F_v/F_m of the tropical-origin C₃ species were less sensitive to 45 °C treatment, regardless of a large variation of T_c; thus no significant correlation was found between their RF_v/m and T_c. Thus T_c might not be a suitable index of heat tolerance for plants with wide range of environmental adaptation. Nevertheless, T_c's of tropical origin C₃ species, varying and showing high plasticity to seasonal changes and temperature treatment, appeared suitable for the estimation of the degree of temperature acclimation in the same species.     

Assessment of the frost sensitivity of Trifolium species by chlorophyll fluorescence analysis

The potential of the chlorophyll fluorescence technique in screening for frost sensitivity in a range of Trifolium species from different geographical origins was assessed by measuring the decrease in variable chlorophyll fluorescence (F_var) of leaves after freezing at - 5°C for 60 min. The method was rapid and the results obtained agreed well with a visual assessment of freezing injury carried out after leaves were returned to optimal growth conditions for 72 h. Trifolium alexandrinum (Berseem clover) cv. Tabor originating from Israel was shown to be the most frost sensitive species studied and Trifolium subterraneum (subterranean clover) cv. Mt. Barker, from temperate regions of Australia, the most frost resistant. On extended periods of freezing, frost damage increased and this was associated with a further reduction in variable chlorophyll fluorescence and in quenching capacity of the thylakoid membranes. These results thus indicate that substantial thylakoid membrane dysfunction is induced at freezing temperatures. Furthermore, it was found that frost hardening of the frost sensitive species T. alexandrinum for 21 days at 5°C reduced the extent of damage sustained by the thylakoid membranes as shown by higher fluorescence quenching capacity, smaller reduction in variable fluorescence (F_var) and higher initial fluorescence (F_o) when leaves of hardened plants were frozen at -5°C and -7°C.     

Use of chlorophyll fluorescence to estimate the effect of photoinhibition in outdoor cultures ofSpirulina platensis

Chlorophyll fluorescence measurements were used to evaluate the effect of temperature on photoinhibition inSpirulina platensis cultures grown in tubular reactors outdoors. Cultures grown at 35 °C during the day time showed a lower reduction in the Fv/Fm ratio as compared to cultures grown at 25 °C. It is demonstrated that the lower temperature photoinhibited cells can undergo a complete recovery once transferred to low light and higher temperature. This recovery does not take place when 100 µg ml^-1 chloramphenicol is added to cells. The recovery is light dependent and cells incubated in the dark at low temperature do not show a recovery in the Fv/Fm ratio. The data presented strongly support the hypothesis that photoinhibition takes place in outdoorSpirulina cultures. At the same time it is demonstrated that fluorescence measurements can be used as a fast reliable indication for photoinhibition in outdoor algal cultures.Author for correspondencePublication No. 69 of the Microalgal Biotechnology Laboratory.     

The size of the population of weakly coupled chlorophyll pigments involved in thylakoid photoinhibition determined by steady-state fluorescence spectroscopy

On the basis of experiments with singlet quenchers and in agreement with previous data, it is suggested that a population of energetically weakly coupled chlorophylls may play a central role in photoinhibition in vivo and in vitro. In the present study, we have used steady state fluorescence techniques to gain direct evidence for these uncoupled chlorophylls. Due to the presence of their emission maxima, near 650 nm and more prominently in the 670-675 nm interval both chlorophylls b and a seem to be involved. A straightforward mathematical model is developed to describe the data which allows us to conclude that the uncoupled/weakly coupled population size is in the range of 1-3 molecules per photosystem.     

The size of the population of weakly coupled chlorophyll pigments involved in thylakoid photoinhibition determined by steady-state fluorescence spectroscopy

On the basis of experiments with singlet quenchers and in agreement with previous data, it is suggested that a population of energetically weakly coupled chlorophylls may play a central role in photoinhibition in vivo and in vitro. In the present study, we have used steady state fluorescence techniques to gain direct evidence for these uncoupled chlorophylls. Due to the presence of their emission maxima, near 650 nm and more prominently in the 670--675 nm interval both chlorophylls b and a seem to be involved. A straightforward mathematical model is developed to describe the data which allows us to conclude that the uncoupled/weakly coupled population size is in the range of 1--3 molecules per photosystem.     

ΔpH-dependent chlorophyll fluorescence quenching indicating a mechanism of protection against photoinhibition of chloroplasts

Intact isolated spinach chloroplasts were subjected to photoinhibitory conditions (high light and lack of CO₂). Photoinhibition of the electron transport system was considerably diminished when the chloroplasts were in a low-fluorescent state related to a high proton gradient across the thylakoid membranes, as compared to a high-fluorescent state in which ΔpH-dependent fluorescence quenching was abolished by addition of uncouplers. The hypothesis is discussed that in chloroplasts exposed to excess light, photoinhibition is partly prevented by increased thermal dissipation of excitation energy, as expressed by ΔpH-dependent (‘energy-dependent’) chlorophyll a fluorescence quenching.     

A study on the energy-dependent quenching of chlorophyll fluorescence by means of photoacoustic measurements

The mechanism of energy-dependent quenching (q_E) of chlorophyll fluorescence was studied employing photoacoustic measurements of oxygen evolution and heat release. It is shown that concomitant to the formation of q_E the yield of open reaction centers _p decreases indicating that q_E quenching originates from a process being competitive to fluorescence as well as to photochemistry. The analysis of heat release (rate of thermal deactivation) shows: 1. The competitive process is not given by a still unknown energy storing process. 2. If the competitive process would be a futile cycle the life-times of the involved intermediates had to be faster than 50 s.The results of the photoacoustic measurements are in line with the idea that q_E quenching originates from an increased probability of thermal deactivation of excited chlorophylls.Abbreviations F actual fluorescence - F_m fluorescence yield with all PS II reaction centers closed in a light adapted state - F₀ fluorescence yield with all PS II reaction centers open in a light adapted state - PS Photosystem - _p intrinsic photochemical yield - q_E energy-dependent quenching - q_I photoinhibition quenching - q_N non-photochemical quenching - q_P photochemical quenching - q_T state transition quenching     

Whole-genome analysis of giraffe supports four distinct species

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The response of seedlings of two dipterocarp species to nutrient additions and ectomycorrhizal infection

An experiment was conducted to investigate the effect of ectomycorrhizal infection on growth and nutrient uptake, especially of P and K of dipterocarp seedlings.Hopea helferi (Dyer) Blanco andHopea odorata Roxb. seedlings were grown in a sandy loam soil given a basal dressing. Nutrient treatments were unamended soil (NIL), amended soil with the addition of P and K (F), amended soil without P but with K (-P), amended soil without K but with P (-K), amended soil without P or K addition (-PK). Seedlings grown in the amended soil treatments showed foliar symptoms suggestive of calcium deficiency. Ectomycorrhizal infection appeared to improve shoot Ca concentration and relieved the foliar symptoms. Ectomycorrhizal infection inH. odorata plants increased shoot P concentration and increased shoot and total dry weight to the same or greater extent than those of uninfected plants growing on P amended soil.H. helferi showed a positive response to ectomycorrhizal infection in shoot, root and total dry weight in all nutrient treatments but no response to the nutrient treatments themselves.     

5种松属树种光合功能及叶绿素快相荧光动力学特征比较

以松属5种代表树种日本五针松、华山松、白皮松、油松和赤松为研究对象,比较了它们针叶的形态特征、光合速率和叶绿素快相荧光动力学方面的异同.结果表明:5种松树的针叶形态特征存在显著差异,油松的针叶长度和密度最大;白皮松的叶绿素含量最高;但赤松的光合速率最大,日本五针松的光合速率最小.5种松树针叶的光合速率与气孔导度呈正相关.5种松树的叶绿素快速荧光诱导动力学曲线的差异主要表现在J相和I相.尽管5种松树光系统Ⅱ(PSⅡ)最大光化学效率(Fv/Fm)、潜在光化学活性(Fv/Fo)以及到达最大荧光所需的时间(Tfm)差异不显著,但日本五针松的反应中心关闭净速率dV/dto和dVG/dto以及J相和I相的相对可变荧光强度(Vj和Vi)显著大于其他4种松树,能量分配比率Ψo、φEo、φRo显著低于其他4种松树.这可能与日本五针松的质体醌库大小(Sm)、质体醌库还原速率(Sm/Tfm)和质体醌A(QA)的周转次数(N)最小有关.日本五针松的性能指数PI(ABS/CSo/CSm)和光合驱动力(DF)均显著低于其他4种松树,而赤松PI和DF最高.其中PI(Cso)和PI(CSm)与5种松树的净光合速率呈显著正相关,可以作为评价松树光合能力的荧光指标.     

Ozone sensitivity in herbaceous species as assessed by direct and modulated chlorophyll fluorescence techniques

Seven plant species were exposed in open-top chambers to four levels of ozone (O₃) during two growing seasons and screened for treatment effects on the fast chlorophylla(Chl) fluorescence transient kinetics of dark-adapted leaves, and on the fluorescence signals obtained from the same leaves in illuminated steady-state. The aim was to identify the nature of O₃ effects on PSII, and to determine inter-specific differences. In dark-adapted leaves, O₃ caused a reduction in variable fluorescence (F_V : F₀), indicating an overall reduction in the efficiency of primary photochemistry. A large increase in excitation energy dissipation per active reaction centre (DI₀/RC) and a smaller increase in the trapping rate of excitons (TR₀/RC), showed that a fraction of the reaction centres was inactivated while the rest sustained full functionality. The magnitude of the effect increased in the order ofBromus erectusCentaurea jacea Trisetum flavescens Rumex obtusifolius Plantago lanceolataTrifolium pratense Knautia arvensis. The inter-specific variability in PSII responses could not be explained solely by specific differences in modelled O₃ uptake by the leaves. Visible leaf injury was not related to changes in fluorescence emission. In illuminated steady-state, O₃ sensitivity was most expressed in the change in quantum yield of photosynthetic electron transport (Φ_PSII). The ranking of species differed from the ranking obtained in dark-adapted leaves. These results suggest that the mechanistic basis for O₃ effects on PSII is similar in all species, but that inter-specific differences exist in the magnitude of change which cannot be explained solely by different O₃ uptake rates. The observed changes in fluorescence signals are not O₃-specific.     

Differences in pigment composition, photosynthetic rates and chlorophyll fluorescence images of sun and shade leaves of four tree species.

The differential pigment composition and photosynthetic activity of sun and shade leaves of deciduous (Acer pseudoplatanus, Fagus sylvatica, Tilia cordata) and coniferous (Abies alba) trees was comparatively determined by studying the photosynthetic rates via CO(2) measurements and also by imaging the Chl fluorescence decrease ratio (R(Fd)), which is an in vivo indicator of the net CO(2) assimilation rates. The thicker sun leaves and needles in all tree species were characterized by a lower specific leaf area, lower water content, higher total chlorophyll (Chl) a+b and total carotenoid (Cars) content per leaf area unit, as well as higher values for the ratio Chl a/b compared to the much thinner shade leaves and needles that possess a higher Chl a+b and Cars content on a dry matter basis and higher values for the weight ratio Chls/Cars. Sun leaves and needles exhibited higher rates of maximum net photosynthetic CO(2) assimilation (P(Nmax)) measured at saturating irradiance associated with higher maximum stomatal conductance for water vapor efflux. The differences in photosynthetic activity between sun and shade leaves and needles could also be sensed via imaging the Chl fluorescence decrease ratio R(Fd), since it linearly correlated to the P(Nmax) rates at saturating irradiance. Chl fluorescence imaging not only provided the possibility to screen the differences in P(N) rates between sun and shade leaves, but in addition permitted detection and quantification of the large gradients in photosynthetic rates across the leaf area existing in sun and shade leaves.     

Intra-leaf gradients of photoinhibition induced by different color lights: implications for the dual mechanisms of photoinhibition and for the application of conventional chlorophyll fluorometers

? We studied how different color lights cause gradients of photoinhibition within a leaf, to attempt to resolve the controversy of whether photon absorption by chlorophyll or by manganese (Mn) is the primary cause of photoinhibition, as suggested by the excess-energy hypothesis or the two-step hypothesis, respectively. ? Lincomycin-treated leaf discs were photoinhibited by white, blue, green or red light. Combining a microfiber fluorometer, a fiber-thinning technique and a micro-manipulator enabled us to measure the chlorophyll fluorescence signals within a leaf. Photoinhibition gradients were also compared with results from various conventional fluorometers to estimate their depth of signal detection. ? The severity of photoinhibition was in the descending order of blue, red and green light near the adaxial surface, and in the descending order of blue, green and red light in the deeper tissue, which correlated with the chlorophyll and the Mn absorption spectrums, respectively. These results cannot be explained by either hypothesis alone. ? These data strongly suggest that both the excess-energy and the two-step mechanisms occur in photoinhibition, and fluorometers with red or blue measuring light give overestimated or underestimated F(v)/F(m) values of photoinhibited leaves compared with the whole tissue average, respectively; that is, they measured deeper or shallower leaf tissue, respectively.     

Survivorship and growth of seedlings of four dipterocarp species in a tropical rainforest of Sarawak,East Malaysia

Survivorship and growth of seedlings of four dipterocarp species (Dipterocarpus actangulus, D. globosus, Dryobalanops aromatica, Dryobalanops lanceolata) were studied for 2.5 years in a mixed dipterocarp forest in Sarawak, East Malaysia. Predispersal seed predation rates were larger forD. globosus (75%) thanD. lanceolata (27–34%) andD. aromatica (18–26%). Less than 20% of the twoDryobalanops seeds were damaged by vertebrates after seed dispersal. During the period from seed dispersal to the time when the seedlings had shed cotyledons, more dispersed seeds died in the twoDipterocarpus (ca 90%) than the twoDryobalanops (ca 60–70%). The major mortality factors during this period were uprooted and seed/seedling predation by insects or vertebrates. After the seedlings shed cotyledons, all species showed constant mortality rates of 34, 15–16, 17 and 6%/year forD. actangulus, D. lanceolata, D. aromatica andD. globosus, respectively, in the forest understorey. Mortality was lower in less shaded conditions than in more shaded ones forD. aromatica andD. actangulus, but not significantly different forD. lanceolata andD. globosus. A majority of dead seedlings were killed by fallen branches or were found standing with wilted leaves, probably due to water stress. No significant correlation was found between seed/seedling mortality and distance from mother trees or the initial density of seeds/seedings for all species. The mean leaf production was positively correlated with the estimated diffuse light factor of their habitats for each species.     

Energetics of Photosystem II charge recombination in Acaryochloris marina studied by thermoluminescence and flash-induced chlorophyll fluorescence measurements

We studied the charge recombination characteristics of Photosystem II (PSII) redox components in whole cells of the chlorophyll (Chl) d-dominated cyanobacterium, Acaryochloris marina, by flash-induced chlorophyll fluorescence and thermoluminescence measurements. Flash-induced chlorophyll fluorescence decay was retarded in the mus and ms time ranges and accelerated in the s time range in Acaryochloris marina relative to that in the Chl a-containing cyanobacterium, Synechocystis PCC 6803. In the presence of 3-(3,4-dichlorophenyl)-1, 1-dimethylurea, which blocks the Q(B) site, the relaxation of fluorescence decay arising from S(2)Q(A)(-) recombination was somewhat faster in Acaryochloris marina than in Synechocystis PCC 6803. Thermoluminescence intensity of the so called B band, arising from the recombination of the S(2)Q(B)(-) charge separated state, was enhanced significantly (2.5 fold) on the basis of equal amounts of PSII in Acaryochloris marina as compared with Synechocystis 6803. Our data show that the energetics of charge recombination is modified in Acaryochloris marina leading to a approximately 15 meV decrease of the free energy gap between the Q(A) and Q(B) acceptors. In addition, the total free energy gap between the ground state and the excited state of the reaction center chlorophyll is at least approximately 25-30 meV smaller in Acaryochloris marina, suggesting that the primary donor species cannot consist entirely of Chl a in Acaryochloris marina, and there is a contribution from Chl d as well.     

Insights on the development, kinetics, and variation of photoinhibition using chlorophyll fluorescence imaging of a chilled, variegated leaf

The effect of chilling on photosystem II (PSII) efficiency was studied in the variegated leaves of Calathea makoyana, in order to gain insight into the causes of chilling-induced photoinhibition. Additionally, a relationship was revealed between (chilling) stress and variation in photosynthesis. Chilling treatments (5 degrees C and 10 degrees C) were performed for different durations (1-7 d) under a moderate irradiance (120 micromol m-2 s-1). The individual leaves were divided into a shaded zone and two illuminated, chilled zones. The leaf tip and sometimes the leaf base were not chilled. Measurements of the dark-adapted Fv/Fm were made on the different leaf zones at the end of the chilling treatment, and then for several days thereafter to monitor recovery. Chilling up to 7 d in the dark did not affect PSII efficiency and visual appearance, whereas chilling in the light caused severe photoinhibition, sometimes followed by leaf necrosis. Photoinhibition increased with the duration of the chilling period, whereas, remarkably, chilling temperature had no effect. In the unchilled leaf tip, photoinhibition also occurred, whereas in the unchilled leaf base it did not. Whatever the leaf zone, photoinhibition became permanent if the mean value dropped below 0.4, although chlorosis and necrosis were associated solely with chilled illuminated tissue. Starch accumulated in the unchilled leaf tip, in contrast to the adjacent chilled irradiated zone. This suggests that photoinhibition was due to a secondary effect in the unchilled leaf tip (sink limitation), whereas it was a direct effect of chilling and irradiance in the chilled illuminated zones. The PSII efficiency and its coefficient of variation showed a unique negative linearity across all leaf zones and different tissue types. The slope of this curve was steeper for chilled leaves than it was for healthy, non-stressed leaves, suggesting that the coefficient of variation may be an important tool for assessing stress in leaves.