气体交换与荧光同步测量估算植物光合电子流的分配

光合电子流分配是植物光合控制的一个重要环节。然而,传统电子流分配的计算方法存在诸多问题尚未引起人们的注意,如:(1)低估了光呼吸每释放一个CO2分子所消耗的电子数;(2)混淆了相对电子传递速率和绝对电子传递速率;(3)忽略了除碳同化和光呼吸外的其他电子流分配途径;(4)难以准确获取光下暗呼吸速率值,从而导致碳同化电子流(JC)及光呼吸速率(Rp)的不准确估算等。以小麦和大豆气体交换与荧光同步测量数据为例,结果表明大豆电子传递速率与碳同化两者对光强的响应一致性较好,同时达到最大值;而小麦的一致性相对较差,说明电子传递速率和碳同化并非完全一致,推测认为有可能与作物对同化产物输出的模式不同有关。通过光呼吸速率换算出的电子流(12×Rp)与实际测量电子流(ΔJO)之间存在较大的差异;另外,传统方法估算出的光呼吸速率(估算值)与光呼吸测量值之间也存在较大差异,分析认为这主要是由于绝对光合速率与相对电子传递速率之间差异造成。     

Nitrogen application improves gas exchange characteristics and chlorophyll fluorescence in maize hybrids under salinity conditions

The understanding of crop physiological responses to salinity stress is of paramount importance for selection of genotypes with improved tolerance to this stress. Maize (Zea mays L.) hybrids Pioneer 32B33 and Dekalb 979 were grown in pots and subjected to three levels of salinity under four nitrogen levels to determine the role of nitrogen under saline conditions. Salinity stress effects on gas exchange characteristics and chlorophyll fluorescence of maize hybrids were evaluated under semi-controlled conditions. Under salinity stress, the changes in the net photosynthetic rate (P _N), stomatal conductance (g _s), and transpiration rate (E) were similarly directed: all decreased and were lower than in control at the higher salinity level (10 dS/m). Water use efficiency was increased with increasing salinity since transpiration was stronger depressed by salt than photosynthesis. Plants subjected to the lower level of salinity did not differ from control in tested characteristics. Nitrogen application ameliorated the effects of salinity.     

5-Aminolevulinic acid enhances photosynthetic gas exchange, chlorophyll fluorescence and antioxidant system in oilseed rape under drought stress

This study evaluates the role of exogenous foliar application of 5-aminolevulinic acid (ALA) on water relations, gas exchange, chlorophyll fluorescence, and the activities and gene expression patterns of antioxidant enzymes in leaves of oilseed rape under drought stress and recovery conditions. Seedlings at four-leaf stage were imposed to well-watered condition (80 % of water-holding capacity) or drought stress (40 % of water-holding capacity) and subsequently foliar sprayed with water or ALA (30 mg l^?1). Drought suppressed the accumulation of plant biomass and decreased chlorophyll content and leaf water status (relative water content and water potential). The actual quantum yield of photosystem II and electron transport rates were hampered in parallel to net photosynthetic rate. However, drought stress induced the accumulation of malondialdehyde (MDA) and hydrogen peroxide, enhanced the activities of catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX), glutathione reductase (GR) and superoxide dismutase and up-regulated the expression of APX and GR. After rehydration for 4 days, the growth of drought-treated seedlings was restored to normal level for most of the physiological parameters. Foliar application of ALA maintained relatively higher leaf water status and enhanced chlorophyll content, net photosynthetic rate, actual quantum yield of photosystem II, photochemical quenching, non-photochemical quenching and electron transport rates in stressed leaves. Exogenous ALA also alleviated the accumulation of MDA and hydrogen peroxide, increased the activities of antioxidant enzymes and enhanced the expression of CAT and POD in drought-treated plants. These results indicate that ALA may effectively protect rapeseed seedlings from damage induced by drought stress.     

Deriving C4 photosynthetic parameters from combined gas exchange and chlorophyll fluorescence using an Excel tool: theory and practice

The higher photosynthetic potential of C₄ plants has led to extensive research over the past 50 years, including C₄‐dominated natural biomes, crops such as maize, or for evaluating the transfer of C₄ traits into C₃ lineages. Photosynthetic gas exchange can be measured in air or in a 2% Oxygen mixture using readily available commercial gas exchange and modulated PSII fluorescence systems. Interpretation of these data, however, requires an understanding (or the development) of various modelling approaches, which limit the use by non‐specialists. In this paper we present an accessible summary of the theory behind the analysis and derivation of C₄ photosynthetic parameters, and provide a freely available Excel Fitting Tool (EFT), making rigorous C₄ data analysis accessible to a broader audience. Outputs include those defining C₄ photochemical and biochemical efficiency, the rate of photorespiration, bundle sheath conductance to CO₂ diffusion and the in vivo biochemical constants for PEP carboxylase. The EFT compares several methodological variants proposed by different investigators, allowing users to choose the level of complexity required to interpret data. We provide a complete analysis of gas exchange data on maize (as a model C₄ organism and key global crop) to illustrate the approaches, their analysis and interpretation. © 2015 John Wiley & Sons Ltd     

Using combined measurements of gas exchange and chlorophyll fluorescence to investigate the photosynthetic light responses of plant species adapted to different light regimes

One broad-leaved pioneer tree, Alnus formosana, two broad-leaved understory shrubs, Ardisia crenata and Ardisia cornudentata, and four ferns with different light adaptation capabilities (ranked from high to low, Pyrrosia lingus, Asplenium antiquum, Diplazium donianum, Archangiopteris somai) were used to elucidate the light responses of photosynthetic rate and electron transport rate (ETR). Pot-grown materials received up to 3 levels of light intensity, i.e., 100%, 50% and 10% sunlight. Both gas exchange and chlorophyll (Chl) fluorescence were measured simultaneously by an equipment under constant temperature and 7 levels (0?C2,000 ??mol m^?2 s^?1) of photosynthetic photon flux density (PPFD). Plants adapted to-or acclimated to high light always had higher light-saturation point and maximal photosynthetic rate. Even materials had a broad range of photosynthetic capacity [maximal photosynthetic rate ranging from 2 to 23 ??mol(CO₂) m^?2 s^?1], the ratio of ETR to gross photosynthetic rate (P _G) was close for A. formosana and the 4 fern species when measured under constant temperature, but the PPFD varied. In addition, P. lingus and A. formosana grown under 100% sunlight and measured at different seasonal temperatures (15, 20, 25, and 30°C) showed increased ETR/P _G ratio with increasing temperature and could be fitted by first- and second-order equations, respectively. With this equation, estimated and measured P _G were closely correlated (r ² = 0.916 and r ² = 0.964 for P. lingus and A. formosana, respectively, p<0.001). These equations contain only the 2 easily obtained dynamic indicators, ETR and leaf temperature. Therefore, for some species with near ETR/P _G ratio in differential levels of PPFD, these equations could be used to simulate dynamic variation of leaf scale photosynthetic rate under different temperature and PPFD conditions.     

Effects of drought stress on photosynthetic gas exchange, chlorophyll fluorescence and stem diameter of soybean plants

Changes in plant growth, photosynthetic gas exchange, chlorophyll fluorescence and stem diameter of soybean [Glycine max (L.) Merr.] plants under drought stress were studied. Total plant dry mass was reduced by 30 % compared to well-watered control plants. Leaf water potential was slightly decreased by water stress. Water stress induced daytime shrinkage and reduced night-time expansion of stem. Photosynthetic rate, stomatal conductance and transpiration rate were significantly declined by water stress, while the intercellular CO₂ concentration was changed only slightly at the initiation of stress treatment. The maximum photochemical efficiency of photosystem 2 and apparent photosynthetic electron transport rate were not changed by water stress.     

The ontogenetic approach to chlorophyll fluorescence studies of plant photosynthetic apparatus under stressful conditions

Based on the analysis of reasons limiting the application of the method of chlorophyll fluorescence induction for estimating the state of the leaf photosynthetic apparatus under prolonged stress, the necessity of the ontogenetic approach consisting in a more exact determination of leaf age was substantiated. A comparison of the calendar and ontogenetic ways of determination of age of cucumber leaves under controlled conditions revealed essential distinctions in the estimation of plant leaf photosynthetic apparatus by the method of chlorophyll fluorescence induction for two variants distinguishing by the cultivation light regime ("white", 400-700 nm, and "red", 600-700 nm). It was shown that, in the case of prolonged effect of the stress factor on the plant, the unambiguity of the interpretation of chlorophyll fluorescence induction parameters in the estimation of the state of their photosynthetic apparatus depends essentially on the choice of the ontogenetic period of leaves of plants being compared and the accuracy of determination of leaf age.     

Mannose metabolism in corn and its impact on leaf metabolites, photosynthetic gas exchange, and chlorophyll fluorescence

When intact corn leaves were provided millimolar concentrations of d-mannose through the transpiration stream photosynthesis was inhibited; 5.7 millimolar resulted in a 50% inhibition of the carbon exchange rate. This inhibition was partially reversible by the addition of orthophosphate to the feeding solution. Mannose metabolism by corn leaves was limited in that it did not act as a resource for sucrose or starch synthesis. Mannose 6-phosphate accumulated in the leaf tissues and was slowly metabolized by a pathway involving mannose 1-phosphate. Correlated with the mannose-6-phosphate accumulation were decreases in ATP, orthophosphate, sucrose, and phosphoenolpyruvate and increases in starch and maltose. When provided in the transpiration stream mannose had access to both mesophyll and bundle sheath cells. Mannose feeding led to oscillations in steady state chlorophyll fluorescence emission (680 nanometers) and an elimination of the Kautsky effect during fluorescence induction. Pyridoxal 5-phosphate and 2,4-dinitrophenol were found to be inhibitors of CO₂ exchange when provided in the transpiration stream of intact corn leaves. However, Pyridoxal 5-phosphate induced a quenching of steady state fluorescence while 2,4-dinitrophenol led to an increase in fluorescence emission.     

Effect of Cd on growth, photosynthetic gas exchange, and chlorophyll fluorescence of wild and Cd-sensitive mutant rice

Growth, photosynthetic gas exchange, and chlorophyll fluorescence characteristics were investigated in wild type (WT) and Cd-sensitive mutant rice (Oryza sativa L.) plants using 50 μM Cd treatment for 12 d followed by a 3-d recovery. Under Cd stress, net dry mass and pigment contents were significantly lower in the mutant plants than in the WT. The mutant had lower net photosynthetic rate (P _N), transpiration rate (E), and stomatal conductance (g _s) than WT rice, however, it had higher intercellular CO₂ concentration (C _i), indicating that non-stomatal factors accounted for the inhibition of P _N. Maximal photochemical efficiency of photosystem 2 (F_v/F_m), effective quantum yield of PS2 (Φ_PS2), and photochemical quenching (q_P) decreased much in the mutant under Cd stress. Cd content in roots and leaves of the mutant was significantly higher than those in the WT. Hence Cd toxicity was associated with the marked increases in Cd contents of plant tissue. After the recovery for 3 d, the WT rice had higher capacity to recover from Cd injury than the mutant.     

An Excel tool for deriving key photosynthetic parameters from combined gas exchange and chlorophyll fluorescence: theory and practice

Combined photosynthetic gas exchange and modulated fluorometres are widely used to evaluate physiological characteristics associated with phenotypic and genotypic variation, whether in response to genetic manipulation or resource limitation in natural vegetation or crops. After describing relatively simple experimental procedures, we present the theoretical background to the derivation of photosynthetic parameters, and provide a freely available Excel‐based fitting tool (EFT) that will be of use to specialists and non‐specialists alike. We use data acquired in concurrent variable fluorescence–gas exchange experiments, where A/C_i and light–response curves have been measured under ambient and low oxygen. From these data, the EFT derives light respiration, initial PSII (photosystem II) photochemical yield, initial quantum yield for CO₂ fixation, fraction of incident light harvested by PSII, initial quantum yield for electron transport, electron transport rate, rate of photorespiration, stomatal limitation, Rubisco (ribulose 1·5‐bisphosphate carboxylase/oxygenase) rate of carboxylation and oxygenation, Rubisco specificity factor, mesophyll conductance to CO₂ diffusion, light and CO₂ compensation point, Rubisco apparent Michaelis–Menten constant, and Rubisco CO₂‐saturated carboxylation rate. As an example, a complete analysis of gas exchange data on tobacco plants is provided. We also discuss potential measurement problems and pitfalls, and suggest how such empirical data could subsequently be used to parameterize predictive photosynthetic models.     

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.     

Photosynthesis of terrestrial cyanobacteria under light and desiccation stress as expressed by chlorophyll fluorescence and gas exchange

Terrestrial mats of cyanobacteria with other associated microscopicalcryptogams were obtained from various sites in the tropics,i.e. rocks of mountains and rock-outcrops and bare soil surfaces,a valley in the Austrian Alps and a glasshouse. Species diversityof each sample was analysed qualitatively. The samples camefrom very different light climates. Responses to light and desiccationstress were studied using the saturation pulse method for recordingchlorophyll fluorescence variables as well as by measuring netCO² and O² exchange in order to confirm results by independentmethods. Under light stress, shade and high-light, samples showeda reduction of gas exchange and of the fluorescence variablesphotochemical fluorescence quenching coefficient (qp), potentialquantum yield of photosystem II (F_vIF^m) and effective quantumyield (     

The responses of photosynthetic gas exchange and chlorophyll a fluorescence to changes of irradiance and temperature in two species of Miscanthus

Photosynthetic CO₂ uptake and chlorophyll (Chl) a fluorescence of C₄ perennial grasses, Miscanthus floridulus (Labill) Warb and M. transmorrisonensis Hayata, from altitudes in central Taiwan of 390 and 2700 m, respectively, were studied at 10 and 25 °C to find if the species differ in their photosynthetic responses to a low temperature, and whether their photosystems 2 become more susceptible to the photoinhibition at low temperatures. For both species, the maximum photosynthetic rate (P_max) was reduced when the leaves were exposed to 10 °C. At irradiances higher than 400 µmol m^-2 s^-1, the values of F_v/F_m were reduced in both species at 10 °C but not at 25 °C, which indicated the photoinhibition at 10 °C. Reductions in P_max and the values of F_v/F_m at 10 °C were lesser in M. transmorrisonensis than in M. floridulus.     

Beneficial role of acetylcholine in chlorophyll metabolism and photosynthetic gas exchange in Nicotiana benthamiana seedlings under salinity stress

• Acetylcholine (ACh) is believed to improve plant growth. However, regulation at biochemical and molecular levels is largely unknown.
• The present study investigated the impact of exogenously applied ACh (10 µm ) on growth and chlorophyll metabolism in hydroponically grown Nicotiana benthamiana under salt stress (150 mm NaCl).
• Salinity reduced root hydraulic conductivity while ACh‐treated seedlings exhibited a significant increase, resulting in increased relative water content. Salinity induced a reduction in chlorophyll biosynthetic intermediates, such as protoporphyrin‐IX, Mg‐photoporphyrin‐IX and protochlorophyllide, which were significantly ameliorated in the presence of ACh. This influence of ACh on chlorophyll synthesis was confirmed by up‐regulation of HEMA1, CHLH, CAO and POR genes. Gas exchange parameters, i.e. stomatal conductance, internal CO₂ concentration and transpiration rate, increased with ACh, thereby alleviating the salinity effects on photosynthesis. In addition, the salinity‐induced enhancement of lipid peroxidation declined after ACh treatment through modulation of the activity of the assayed antioxidant enzymes (superoxide dismutase and peroxidase). Importantly, ACh significantly reduced the uptake of Na and increased uptake of K, resulting in a decline in the Na/K ratio.
• Results of the present study indicate that ACh can be effective in ameliorating NaCl‐induced osmotic stress, altering chlorophyll metabolism and thus photosynthesis by maintaining ion homeostasis, hydraulic conductivity and water balance.

     

The responses of photosynthetic gas exchange and chlorophyll a fluorescence to changes of irradiance and temperature in two species of Miscanthus

Photosynthetic CO₂ uptake and chlorophyll (Chl) a fluorescence of C₄ perennial grasses, Miscanthus floridulus (Labill) Warb and M. transmorrisonensis Hayata, from altitudes in central Taiwan of 390 and 2700 m, respectively, were studied at 10 and 25 °C to find if the species differ in their photosynthetic responses to a low temperature, and whether their photosystems 2 become more susceptible to the photoinhibition at low temperatures. For both species, the maximum photosynthetic rate (P_max) was reduced when the leaves were exposed to 10 °C. At irradiances higher than 400 μmol m^-2 s^-1, the values of F_v/F_m were reduced in both species at 10 °C but not at 25 °C, which indicated the photoinhibition at 10 °C. Reductions in P_max and the values of F_v/F_m at 10 °C were lesser in M. transmorrisonensis than in M. floridulus. This revised version was published online in June 2006 with corrections to the Cover Date.     

Influence of foliar-applied triacontanol on growth, gas exchange characteristics, and chlorophyll fluorescence at different growth stages in wheat under saline conditions

A greenhouse experiment was conducted to examine the effect of foliar application of triacontanol (TRIA) on two cultivars (cv. S-24 and MH-97) of wheat (Triticum aestivum L.) at different growth stages. Plants were grown in full strength Hoagland’s nutrient solution under salt stress (150 mM NaCl) or control (0 mM NaCl) conditions. Three TRIA concentrations (0, 10, and 20 μM) were sprayed over leaves at three different growth stages, i.e. vegetative (V), boot (B), and vegetative + boot (VB) stages (two sprays on same plants, i.e., the first at 30-d-old plants and the second 78-d-old plants). Salt stress decreased significantly growth, net photosynthetic rate (P _N), transpiration rate (E), chlorophyll contents (Chl a and b), and electron transport rate (ETR), while membrane permeability increased in both wheat cultivars. Stomatal conductance (g _s) decreased only in salt-sensitive cv. MH-97 under saline conditions. Foliar application of TRIA at different growth stages enhanced significantly the growth, P _N, g _s, Chl a and b contents, and ETR, while membrane permeability was reduced in both cultivars under salt stress. Of various growth stages, foliar-applied TRIA was comparatively more effective when it was applied at V and VB stages. Overall, 10 μM TRIA concentration was the most efficient in reducing negative effects of salinity stress in both wheat cultivars. The cv. S-24 showed the better growth and ETR, while cv. MH-97 exhibited higher nonphotochemical quenching.     

Effects of cytokinin on photosynthetic gas exchange, chlorophyll fluorescence parameters and antioxidative system in seedlings of eggplant (Solanum melongena L.) under salinity stress

The effects of 6-benzyladenine (6-BA) on plant growth, photosynthetic gas exchange, chlorophyll fluorescence and antioxidant systems of eggplant (Solanum melongena L.) under salt stress were investigated. Eggplant seedlings were exposed to 90?mM NaCl with four levels of 6-BA (5, 10, 20 and 50???M) for 10?days. 6-BA at lower concentrations increased chlorophyll concentration, the net photosynthetic rate (P _N), stomatal conductance (g _s), and transpiration rate (E), intercellular CO₂ concentration (C _i) and water use efficiency (WUE), as well as the quantum efficiency of PSII photochemistry (??PSII), photochemical quenching (q _p), and decreased non-photochemical quenching (NPQ), while higher concentrations reduced the effects or even exacerbated the occurrence of photosynthetic capacity. The activities of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) increased significantly during salt treatments, and induced the increase of the activities of these enzymes at certain concentrations of 6-BA. 6-BA also reduced significantly malonaldehyde (MDA) contents and O ₂ ^·? production. It was concluded that 6-BA could alleviate the detrimental effects of salt stress on plant growth by increasing photosynthetic efficiency and enhancing antioxidant enzyme systems in leaves at a proper concentration and of the varying 6-BA concentrations used, the most effective concentration for promoting growth was 10???M under saline conditions.     

土壤盐分对棉花功能叶气体交换参数和叶绿素荧光参数日变化的影响

2007—2008年在南京农业大学牌楼试验站进行盆栽试验,选择耐盐品种中棉所44和盐敏感品种苏棉12号为材料,试验设置5个土壤盐分水平(0、0.35%、0.60%、0.85%和1.00%),研究土壤盐分对棉花功能叶气体交换参数和叶绿素荧光参数日变化的影响.结果表明:随土壤盐分水平的升高,棉花功能叶中Na+、C l-和Mg2+含量升高,K+和Ca2+含量降低.低于0.35%盐分处理对叶片气体交换参数和叶绿素荧光参数的影响较小,高于0.35%的盐分处理显著降低了棉花功能叶的净光合速率(Pn),提高了棉花功能叶对日间光辐射强度和温度的敏感程度,导致光温抑制现象加重,并改变了Pn和气孔导度(Gs)的日变化趋势,使其由单峰曲线逐渐变为持续下降趋势.随日间光辐射强度和温度的变化,棉花叶片最大光化学效率(Fv/Fm)、光系统Ⅱ(PSⅡ)量子产量(ΦPSⅡ)和光化学猝灭系数(qP)的日变化趋势呈V型曲线,最低值出现在12:00—13:00,非光化学猝灭系数(qN)的日变化趋势呈单峰曲线;盐分处理降低了棉花功能叶Fv/Fm、ΦPSⅡ和qP,提高了qN,且增大了其变化幅度.耐盐品种中棉所44功能叶片中较低的Na+、C l-含量及...     

Effects of cytokinin on photosynthetic gas exchange, chlorophyll fluorescence parameters, antioxidative system and carbohydrate accumulation in cucumber (Cucumis sativus L.) under low light

The effects of 6-benzylaminopurine (6-BA) on plant growth, net photosynthetic rate, relative chlorophyll content, soluble protein, carbohydrates contents and antioxidant systems of cucumber (Cucumis sativus L.) under low-light environment were investigated using two different cucumber cultivars. The results showed that the weak light resulted in the remarkable decrease in plant net photosynthetic rate, relative chlorophyll content, soluble protein and carbohydrates contents, but promoted the superoxide dismutase and guaiacol peroxidase activities. However, application of 6-BA alleviated the reduction of the correlative parameters and mediated the changes of antioxidant systems. The potential mechanisms may involve the following aspects: 6-BA clearly enhanced the plants’ tolerance to low light by increasing chlorophyll content, reducing the production of superoxide radical (O ₂ ^·? ), and enhancing the quenching of hydrogen peroxide (H₂O₂), consequently alleviating the injury of photosynthetic system, and further increasing the efficiency of CO₂ assimilation, producing more carbohydrates which can meet the growth need of cucumber. Meanwhile, the present study indicated that cucumber of Europe mini type (Chunqiuwang) was more tolerant to low light than HuaNan type (Huza No.3).