Fluorescence decay studies of reduced nicotinamide adenine dinucleotide in solution and bound to liver alcohol dehydrogenase.

The monophoton counting technique was used to obtain the fluorescence decay kinetics of NADH (dihydronicotinamide adenine dinucleotide) bound to LADH (HORSE LIVER ALCOHOL DEHYDROGENAS). It was found that the fluorescence decay of the enzyme complex did not follow a single exponential decay law but that the data could be well described as a sum of two exponentials. The decay parameters of the enzyme complex do not depend on the degree of binding-site saturation. These results are interpreted in terms of a reversible excited-state reaction forming a nonfluorescent product. Fluorescence decay kinetics are also reported for NADH and related molecules in solution. The decay parameters, fluorescence emission maxima, and fluorescence intensities depend on solvent polarity and viscosity.     

喀斯特干旱环境对青冈栎叶片生长及叶绿素荧光动力学参数的影响

为了探讨喀斯特植被恢复树种青冈栎对干旱环境的适应机制,以当年生青冈栎实生幼苗为材料进行盆栽控水试验,设置正常浇水(-0.1 MPa,对照)、轻度干旱(-0.5 MPa)、中度干旱(-0.9 MPa)和重度干旱(-1.5 MPa)胁迫处理,研究持续干旱处理(15、30、45、60和90天)对其幼苗叶片生长及叶绿素荧光参数的影响.结果表明: 随着干旱胁迫强度的加剧, 叶片的单叶面积、健康叶片数量、叶片含水率、总叶绿素、类胡萝卜素、最大荧光、最大光化学量子产量和潜在光化学效率均显著下降,而枯叶数量和初始荧光显著增加.这些参数在轻度干旱胁迫处理和对照之间均无显著差异.在轻度干旱胁迫处理下,青冈栎幼苗叶片PSⅡ单位反应中心吸收的光能(ABS/RC)、捕获用于还原q_A的能量(TR_o/RC)、单位面积内有活性的反应中心数目(RC/CS)、单位面积捕获的光能(TR_o/CS)和单位面积内用于电子传递的光能(ET_o/CS)均与对照无显著差异,其中RC/CS总是略高于对照,TR_o/CS和ET_o/CS均在第45天达到峰值,分别为606.12和440.78;而中度和重度干旱胁迫处理叶片的ABS/RC、TR_o/RC、ET_o/RC、DIR_o/RC、RC/CS、TR_o/CS和ET_o/CS均低于对照,且随干旱胁迫时间的延长,重度干旱胁迫处理下降更显著.随干旱胁迫的加剧和时间的延长,叶片最大量子效率、其他电子受体的概率和电子传递的量子比率均下降,而用于热耗散的量子比率增加.轻度干旱胁迫下青冈栎幼苗表现出较强的适应性,中度干旱胁迫引起部分叶绿素荧光参数和光合色素指标下降,导致幼苗生长缓慢,而重度干旱则对幼苗生长的影响较为严重,但幼苗未出现死亡现象.因此,青冈栎幼苗有较强的干旱忍受能力,适合在喀斯特地区植被恢复重建和造林工程中应用.     

Determination of DNA polymerase and nuclease activities of DNA-dependent polymerases using fluorescence detection under real-time conditions

A new method is proposed for estimation of polymerase activities using fluorescence detection during isothermal reaction. The method allows simultaneous determination of DNA-dependent DNA polymerase and 5'-3'-exonuclease activities using amplifiers supplied with an optical module for fluorescence detection under real-time conditions. Different primer-template combinations used as polymerase substrates were compared. Primer elongation (polymerase reaction) is detected by changes in SYBR Green I fluorescence upon binding to dsDNA during reaction; nuclease activities are detected by changes in fluorescence due to cleavage of the probe, containing the reporter fluorophore and fluorescence quencher, and hybridized in advance to the template single-stranded region. It was also shown that the method can be used for determination of relative activities of DNA polymerase preparations, estimation of temperature-time dissociation parameters of polymerase complexes with specific antibodies to its active center, and analysis of effects of inhibitors and activators of different nature on reaction rates of dsDNA polymerization and 5'-3'-exonuclease cleavage by polymerase. The method can be also used for estimation of endonuclease activities of DNA polymerases.     

快速叶绿素荧光诱导动力学分析在光合作用研究中的应用

JIP-测定(JIP-test)是以生物膜能量流动为基础建立的分析方法.利用该方法可以获得有关光系统Ⅱ的大量信息.文章介绍了快速叶绿素荧光诱导动力学曲线的定义、数据分析方法及相关参数的意义,并举例说明如何利用该方法分析不同环境条件对光合机构主要是PSⅡ的供体侧、受体侧及PSⅡ反应中心的影响.     

Effects of gamma-aminobutyric acid on the photosynthesis and chlorophyll fluorescence parameters of muskmelon seedlings under hypoxia stress

By the method of hydroponic culture, this paper studied the effects of exogenous gamma-aminobutyric acid (GABA) on the photosynthetic pigment contents, photosynthesis, and chlorophyll fluorescence parameters of muskmelon seedlings under hypoxia stress. Hypoxia stress induced a significant decrease of photosynthetic pigment contents, resulting in the decrease of photosynthesis. Applying GABA could significantly increase the photosynthetic pigment contents, net photosynthetic rate (P(n)), stomatal conductance (G(s)), intercellular CO2 concentration (C(i)), carboxylation efficiency (CE), maximal photochemical efficiency of PS II (F(v)/F(m)), photochemical quenching (q(P)), apparent photosynthetic electron transfer rate (ETR), and quantum yield of PS II electron transport (phi(PS II)), and decrease the stomatal limitation value (L(s)), minimal fluorescence (F(o)), and non-photochemical quenching (NPQ) under both hypoxic and normal conditions. The alleviation effect of GABA on photosynthetic characteristics was more obvious under hypoxia stress. However, simultaneously applying GABA and VGB could significantly decrease the alleviation effect of GABA under hypoxia stress.     

光叶眼子菜响应夏季高温的模拟研究

为探讨南四湖优势物种光叶眼子菜在夏季浅水区的衰亡原因, 用25℃、30℃、35℃和40℃的恒温水浴模拟夏季高温处理光叶眼子菜(Co. Potamogeton lucens L.)3h。生化结果显示, 在35℃及以上高温下, 光叶眼子菜的蛋白质含量、可溶性糖含量和叶绿素含量显著下降, 丙二醛含量显著上升, 说明35℃以上高温对光叶眼子菜产生了显著伤害。光叶眼子菜的光合系统对高温更为敏感, 在高温胁迫下标准化的叶绿素荧光动力学曲线上J相和K相显著隆起, 但并未发现明显的L-band。进一步解析叶片的叶绿素荧光动力学参数, 结果显示: 随着处理温度的升高, 反应中心的初始关闭速率(dVG/dt_o, dV/dt_o)变慢, 但到达P相的所需时间(T_fm)变短; 光系统Ⅱ (Photosystem Ⅱ, PSⅡ)的光化学效率(F_v/F_m)减小, 非光化学效率(K_n)、J相相对可变荧光强度(V_j)和热耗散(DI_o/RC、DI_o/CS_o、F_o/F_m)增大; 尽管高温下质体醌周转次数(N)、还原速率(S_m/T_fm)和I相相对可变荧光强度(V_i)变化不显著, 但质体醌库(S_m)明显减小; 单个反应中心光能的吸收(ABS/RC)和捕获效率(TR_o/RC)增加, 电子传递效率(ET_o/RC)却呈下降趋势; 单位激发态面积的光能捕获(TR_o/CS_o)和电子传递效率(ET_o/CS_o)均降低, 反应中心数目(RC/CS_o)显著减少。上述高温胁迫效应导致整个叶片的结构功能指数(SFI_abs)、性能指数(PI_abs)以及光合驱动力(DF)显著降低。高温对光叶眼子菜的伤害主要是导致其光系统II放氧复合体失活、反应中心数目减少和反应中心的光化学效率下降, 进而诱导活性氧的产生, 对细胞造成伤害。因此, 光叶眼子菜属于对高温敏感的水生植物。     

NaCl胁迫加重强光胁迫下超大甜椒叶片的光系统II和光系统I的光抑制

快速叶绿素荧光动力学可以在无损情况下探知叶片光合机构的损伤程度, 快速叶绿素荧光测定和分析技术(JIP-test)将测量值转化为多种具有生物学意义的参数, 因而被广泛应用于植物光合机构对环境的响应机制研究。该文研究了超大甜椒(Capsicum annuum)幼苗在强光及不同NaCl浓度胁迫下的荧光响应情况。与单纯强光胁迫相比, NaCl胁迫引起了叶绿素荧光诱导曲线的明显改变, 光系统II (PSII)光抑制加重, 同时PSII反应中心和受体侧受到明显影响, 而且高NaCl浓度胁迫下PSII供体侧受伤害明显, 同时PSI反应中心活性(P700⁺)在盐胁迫下明显降低。这些结果表明, NaCl胁迫会增强强光对超大甜椒光系统的光抑制, 并且浓度越高抑制越明显, 但对PSI的抑制作用低于PSII。高NaCl浓度胁迫易对PSII供体侧造成破坏, 且PSI光抑制严重。     

Response of the photosynthetic apparatus of cotton (Gossypium hirsutum) to the onset of drought stress under field conditions studied by gas-exchange analysis and chlorophyll fluorescence imaging.

The functioning of the photosynthetic apparatus of cotton (Gossypium hirsutum) grown during the onset of water limitation was studied by gas-exchange and chlorophyll fluorescence to better understand the adaptation mechanisms of the photosynthetic apparatus to drought conditions. For this, cotton was grown in the field in Central Asia under well-irrigated and moderately drought-stressed conditions. The light and CO(2) responses of photosynthesis (A(G)), stomatal conductance (g(s)) and various chlorophyll fluorescence parameters were determined simultaneously. Furthermore, chlorophyll fluorescence images were taken from leaves to study the spatial pattern of photosystem II (PSII) efficiency and non-photochemical quenching parameters. Under low and moderate light intensity, the onset of drought stress caused an increase in the operating quantum efficiency of PSII photochemistry (varphi(PSII)) which indicated increased photorespiration since photosynthesis was hardly affected by water limitation. The increase in varphi(PSII) was caused by an increase of the efficiency of open PSII reaction centers (F(v)'/F(m)') and by a decrease of the basal non-photochemical quenching (varphi(NO)). Using a chlorophyll fluorescence imaging system a low spatial heterogeneity of varphi(PSII) was revealed under both irrigation treatments. The increased rate of photorespiration in plants during the onset of drought stress can be seen as an acclimation process to avoid an over-excitation of PSII under more severe drought conditions.     

模拟酸雨对龙眼叶片气体交换和叶绿素a荧光参数的影响（英文）

 研究了模拟酸雨对龙眼(Dimorcarpus longana)叶片气体交换和叶绿素a荧光参数的影响,结果表明：酸雨胁迫抑制龙眼光合作用,受胁迫叶片光补偿点上升。pH 3.0的酸雨处理6 h后叶片气体交换和叶绿素a荧光参数下降,停止处理后72 h可以恢复。pH 2.5的酸雨处理6 h后净光合速率（Pn）、气孔导度(Cs)、蒸腾速率（Tr）,叶绿素a荧光参数Fv/Fo、Fv/Fm、PSII非环式电子传递的量子产量（ΦPSⅡ）、荧光下降比值（Rfd）、非光化学猝灭系数（qN）和光化学猝灭系数（qP）急剧下降,停     

Physiological response of Pinus halepensis needles under ozone and water stress conditions

The aim of this study was to evaluate how physiological processes of potted Pinus halepensis plants, grown under controlled conditions, were affected by ozone (O₃) and/or water stress, integrating the gas exchange and biochemical data with fluorescence OJIP polyphasic transient data. Plants submitted to only water stress (T₁) and with ozone (T₃) showed a strong decrease in stomatal conductance and gas exchange, coinciding with a reduction of maximum yield of photochemistry ( φ _po) and very negative values of leaf water potential. Simultaneously, a great increase of both PSII antenna size, indicated by absorption per reaction centre, and electron transport per reaction centre were found. The reduction of photosynthesis in the O₃-treated plants (T₂) by a slowing down of the Calvin cycle was supported by the increase of related fluorescence parameters such as relative variable fluorescence, heat de-excitation constant, energy de-excitation by spillover, and the decrease of φ _po. We suggest an antagonistic effect between the two stresses to explain the delayed ozone-induced decrease of stomatal conductance values for T₃ with respect to T₁ plants, by an alteration of the physiological mechanisms of stomatal opening, which involve the increase of intra-cellular free-calcium induced by ABA under co-occurring water shortage. We emphasise the importance of considering the intensity of the individual stress factor in studies concerning the interaction of stresses.     

强光胁迫对濒危植物金花茶幼苗生长和叶绿素荧光参数的影响

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F_o、F_m、F_v、F_v/F_m、F_v/F_o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。     

Variable chlorophyll fluorescence and its use for assessing physiological condition of plant photosynthetic apparatus

Analysis of plant behavior under diverse environmental conditions would be impossible without the methods for adequate assessment of the processes occurring in plants. The photosynthetic apparatus and its reaction to stress factors provide a reliable source of information on plant condition. One of the most informative methods based on monitoring the plant biophysical characteristics consists in detection and analysis of chlorophyll a fluorescence. Fluorescence is mainly emitted by chlorophyll a from the antenna complexes of photosystem II (PSII). However, fluorescence depends not only on the processes in the pigment matrix or PSII reaction centers but also on the redox reactions at the PSII donor and acceptor sides and even in the entire electron transport chain. Presently, a large variety of fluorometers from various manufacturers are available. Although application of such fluorometers does not require specialized training, the correct interpretation of the results would need sufficient knowledge for converting the instrumental data into the information on the condition of analyzed plants. This review is intended for a wide range of specialists employing fluorescence techniques for monitoring the physiological plant condition. It describes in a comprehensible way the theoretical basis of light emission by chlorophyll molecules, the origin of variable fluorescence, as well as relations between the fluorescence parameters, the redox state of electron carriers, and the light reactions of photosynthesis. Approaches to processing and analyzing the fluorescence induction curves are considered in detail on the basis of energy flux theory in the photosynthetic apparatus developed by Prof. Reto J. Strasser and known as a “JIP-test.” The physical meaning and relation of each calculated parameter to certain photosynthetic characteristics are presented, and examples of using these parameters for the assessment of plant physiological condition are outlined.     

Chlorophyll-a fluorescence evaluation of PEG-induced osmotic stress on PSII activity in Arabidopsis plants expressing SIP1

In this study, we evaluated the effect of osmotic stress on photosynthetic machinery of Arabidopsis plants expressing a gene encoding small basic intrinsic protein (SIP1) isolated from Solanum tuberosum. Intact leaves of SIP Arabidopsis plants were exposed to 15% polyethylene glycol (PEG) solution and fast Chlorophyll-a (Chl-a) fluorescence induction kinetics was measured. Photosynthetic parameters like ratio of variable and maximum fluorescence (F_V/F_M), absorbance of photons per active reaction center (ABS/RC), trapping of photons per active reaction center (TRo/RC), electron transport per active reaction center (ETo/RC), and performance index (PI) were measured. Furthermore, the energy pipeline model was deduced in response to PEG stress. The membrane model includes a visualization of the average “antenna size”, which follows the value of the ABS/RC. Analysis of SIP Arabidopsis plants under PEG stress through fast Chl-a fluorescence transient showed that the damage caused due to PEG is more prominent at the donor side rather than the acceptor side of PSII. Higher PI in SIP plants under PEG stress indicated a better vitality than control plants. Overall, these results indicate that constitutive expression of SIP1 in Arabidopsis plants induces significant changes in the photosynthetic machinery under PEG-induced osmotic stress.     

The Effects of Water Stress on Photosystem Ⅱ in Wheat

The fluorescence yield at room temperature, the capacity of excitation energy distribution between photosystem Ⅰ and Ⅱ by Mg2+, variable fluorescence yield, variable fluorescence quenching rate and fluorescence complementary area were decreased under water stress. These indicated that photosystem Il was impaired. The inhibited variable fluorescence yield could be partly recovered by the addition of artificial electron donor DPC. Therefore, water stress inhibited not only the oxidizing site of photosystem Ⅱ but also impaired partly the reaction center of photosystem Ⅱ.     

5-氨基乙酰丙酸对NaCl胁迫下番茄幼苗光合特性的影响

为探讨5-氨基乙酰丙酸（ALA）对NaCl胁迫下番茄光合特性的调控作用,以‘金鹏一号’番茄幼苗为试材,研究叶面喷施50 mg·L^-1或根施10 mg·L^-1 ALA对100 mmol·L^-1 NaCl胁迫下番茄幼苗光合及叶绿素荧光参数的影响.结果表明： NaCl胁迫下,番茄幼苗光合气体交换参数（净光合速率P_n、气孔导度g_s、胞间CO₂浓度C_i、蒸腾速率T_r）及叶绿素荧光参数（实际光化学量子产量F_v′/F_m′、F_m′、PSⅡ反应中心实际光化学效率Φ_PSⅡ、表观光合电子传递效率ETR、光化学淬灭q_P、光化学反应Pc）均显著降低,根施或叶施ALA均可以提高NaCl胁迫下番茄叶片的光合能力,但两种处理方式之间存在一定差异.叶面喷施50 mg·L^-1ALA或根施10 mg·L^-1ALA处理均显著提高了番茄叶片P_n、T_r、g_s和C_i,提高了水分利用效率（WUE）,显著增加了NaCl胁迫下叶片的最大净光合速率,减轻了光抑制.根施ALA对叶绿素含量的作用效果较好,而叶施ALA对光合参数的作用效果较好,两处理叶绿素荧光参数差异不显著.叶面喷施或根施ALA可以提高番茄幼苗的耐盐性,其调控作用与促进叶绿素合成与稳定、维持正常气孔开闭、降低气孔限制,进而提高NaCl胁迫下番茄叶片的光合能力和PSⅡ光化学效率有关.
     

Postharvest Imaging of Chlorophyll Fluorescence from Lemons Can Be Used to Predict Fruit Quality

We demonstrate the feasibility of assaying and predicting post-harvest damage in lemons by monitoring chlorophyll (Chl) fluorescence. Fruit quality was assayed using a commercial instrument that determines photosynthetic performance by imaging Chl fluorescence parameters under different irradiances. Images of Chl fluorescence from individual lemons reveal that photosynthesis is active throughout the post-harvest ripening process. Because photosynthesis is highly sensitive to biotic and abiotic stress, variations in Chl fluorescence parameters over the surface of a lemon fruit can be used to predict areas that will eventually exhibit visible damage. The technique is able to distinguish between mould-infected areas that eventually spread over the surface of the fruit, and damaged areas that do not increase in size during ripening. This study demonstrates the potential for using rapid imaging of Chl fluorescence in post-harvest fruit to develop an automated device that can identify and remove poor quality fruit long before visible damage appears.     

Response of chlorophyll fluorescence parameters to drought stress in sugar beet seedlings

We compared the parameters of chlorophyll fluorescence between two sugar beet (Beta vulgaris L.) species differing in drought tolerance. Our results indicated that there were different responses to the drought stress of these sugar beet species. In drought-tolerant sugar beet, the F ₀ increased slightly, while qN increased substantially, indicating that these plants can protect PSII reaction centers from the damage. F _v/F _m and qP decreased slightly during the initial period of drought stress; this suggests that there is a slight impact of drought stress on the openness of PSII reaction centers, and thus the plants did not suffer seriously. This was further shown by the decreased Yield and electron transfer rate. The parameters of chlorophyll fluorescence were stable and can be used as an important indicator for sugar beet seedlings in the early drought tolerance.     

Effect of oxyanions on the glucose isomerase catalysed equilibrium: 1. Complexes of d-glucose and d-fructose with germanate

The complexing parameters of d-glucose and d-fructose with germanate, derived from various forms of germanium dioxide, have been studied under the conditions pertaining to the d-glucose isomerase (d-xylose isomerase, d-xylose ketol-isomerase, EC 5.3.1.5) reaction. The interaction of germanate with d-glucose and d-fructose at various pH values has been investigated by means of optical rotation methods. The effects of temperature and concentration on the extent of complex formation are reported. The results are used to predict suitable conditions for the enhancement of d-fructose yield in the reaction of d-glucose with this enzyme.     

Chlorophyll Fluorescence as a Tool in Cereal Crop Research

Chlorophyll (Chl) fluorescence is a subtle reflection of primary reactions of photosynthesis. Intricate relationships between fluorescence kinetics and photosynthesis help our understanding of photosynthetic biophysical processes. Chl fluorescence technique is useful as a non-invasive tool in eco-physiological studies, and has extensively been used in assessing plant responses to environmental stress. The review gives a summary of some Chl fluorescence parameters currently used in studies of stress physiology of selected cereal crops, namely water stress, heat stress, salt stress, and chilling stress.     

NaCl胁迫对马齿苋光合作用及叶绿素荧光特性的影响

以马齿苋为材料,采用温室盆栽法研究了14 d NaCl胁迫处理对其幼苗生长、光合作用和叶绿素荧光特性的影响.结果显示:(1)马齿苋幼苗的鲜重和株高在25 mmol·L-1 NaCl胁迫时与对照无显著差异,但其随着NaCl浓度的继续增加均显著降低,且其生物量受到的抑制早于株高.(2) NaCl胁迫下,马齿苋幼苗叶片净光合速率(Pn)降低,胞间二氧化碳浓度(C1)增大,且两者的变化幅度随着NaCl浓度增加而增大.(3)NaCl胁迫下,马齿苋幼苗叶片的初始荧光(F0)、最大荧光(Fm)、可变荧光(Fv)、恒态荧光(Fs)、恒态荧光与初始荧光差值(△F0)、PSⅡ潜在光化学效率(Fv/Fo)和PSⅡ最大光化学效率(Fv/Fm)均降低,叶片光化学荧光猝灭系数(qP)也在NaCl胁迫下降低,而非光化学荧光猝灭系数(NPQ)则上升;在0～50 mmol·L-1NaCl胁迫下,幼苗叶片各荧光参数下降幅度小于其他高浓度NaCl胁迫.研究表明,在NaCl胁迫条件下,马齿苋幼苗叶片的光合作用受光抑制伤害,但在低浓度NaCl下能够较多地将光能用于光化学反应,光抑制程度较低,保持了较高的净光合速率,明显减轻盐胁迫对植株生长的影响,表现出一定的耐盐性.