Soybean Resistance to Phakopsora pachyrhizi as Affected by Acibenzolar‐S‐Methyl,Jasmonic Acid and Silicon

Asian soybean rust (ASR), caused by Phakopsora pachyrhizi, is one of the most important diseases on soybean. At the moment, ASR is managed mainly with fungicides due to the absence of commercial cultivars with resistance to this disease. This study evaluated the effects of acibenzolar‐Smethyl (ASM), jasmonic acid (JA), potassium silicate (PS) and calcium silicate (CS) on soybean resistance to ASR. The ASM, JA and PS were sprayed to leaves 24 h prior to inoculation with P. pachyrhizi. The CS was amended to the soil. The incubation period (time from the inoculation until symptoms development) was longer for plants growing in soil amended with CS or sprayed with ASM in comparison with plants sprayed with water (control). Plants sprayed with ASM had longer latent period (time from the inoculation until signs appearance) in comparison with the control plants. Plants sprayed with PS showed fewer uredia per cm² of leaf in relation to the control plants. The ASM and PS were the most effective treatments in reducing the ASR symptoms in contrast to the JA and CS treatments. The JA served as an inducer of susceptibility to ASR.     

Silicon partially preserves the photosynthetic performance of rice plants infected by Monographella albescens

Leaf scald, caused by Monographella albescens, is one of the major diseases in rice worldwide. This study investigated the effect of silicon (Si) on the photosynthetic gas exchange parameters [net CO₂ assimilation rate (A), stomatal conductance to water vapour (g_s), transpiration rate (E)] and internal CO₂ concentration (C_i), chlorophyll (Chl) fluorescence a parameters [minimal fluorescence (F₀), maximum fluorescence (F_m), maximum quantum yield of photosystem II (F_v/F_m)], photochemical quenching coefficient (q_p), effective quantum yield of PSII [Y(II)], quantum yield of regulated energy dissipation [Y(NPQ)] and quantum yield dissipation non‐regulated [Y(NO)] and the concentrations of pigments in rice plants grown in nutrient solutions containing either 0 (?Si) or 2 mM Si (+Si) and non‐inoculated or inoculated with M. albescens. Leaf scald severity decreased with higher foliar Si concentration. For the inoculated +Si plants, A, g_s and E were significantly higher in comparison with the inoculated ?Si plants, in which C_i was significantly increased. Similarly, the concentrations of Chl_a, Chl_b, total Chl_a+b and carotenoids were higher for the +Si plants in comparison with the ?Si plants. Changes in the images of Chl a fluorescence were first observed precisely on the ?Si plants leaves in comparison with the +Si plants. A decrease of q_P and Y(II) in inoculated ?Si plants, in comparison with the inoculated +Si plants, was accompanied by an increase in Y(NPQ) and Y(NO). Notably, the extent of the leaf areas was much more evident for Y(II) and q_P in comparison with F₀, F_m and F_v/F_m, suggesting that Y(II) and q_P were good predictors in detecting the early effects of leaf scald on the leaf photosynthesis. For the +Si non‐inoculated plants, changes in Y(II) were associated with alterations in both Y(NPQ) and Y(NO) compared with non‐inoculated ?Si plants. In conclusion, the photosynthetic performance (as demonstrated by the gas exchange and Chl a fluorescence parameters) and the pigment pools of rice plants infected with M. albescens were preserved by Si supply and, therefore, provided an increase in rice resistance against leaf scald.     

Glyphosate and nickel differently affect photosynthesis and ethylene in glyphosate-resistant soybean plants infected by Phakopsora pachyrhizi

Nickel (Ni) and glyphosate (Gl) are able to reduce the symptoms of Asian soybean rust (ASR), caused by Phakopsora pachyrhizi, in soybean. However, their combined effects on the energy balance and ethylene metabolism of soybean plants infected with this fungus has not been elucidated. Therefore, the effects of Ni, Gl, and the combination of Ni + Gl on ASR development, photosynthetic capacity, sugar concentrations, and ethylene concentrations in plants of a Gl-resistant cultivar, uninfected or infected with P. pachyrhizi, were investigated. Inoculated plants supplied with Ni had the highest foliar Ni concentration in all the treatments. Gl had a negative effect on the foliar Ni concentration in Ni-sprayed plants. The ASR severity was reduced in plants sprayed with Ni and Gl. Carotenoid and chlorophyll concentrations were higher in inoculated Ni, Gl, and Ni + Gl plants than in control plants. Based on the chlorophyll a fluorescence parameters, the photosynthetic apparatus of the control inoculated plants was damaged, and the least amount of energy was directed to the photochemistry process in these plants. The reduced capacity of the photosynthetic mechanism to capture light and use the energy absorbed by photosystem II in inoculated plants was reflected in their reduced capacity to process CO₂, as indicated by the high internal CO₂ concentrations and low rates of net carbon assimilation. The low sugar concentrations in inoculated plants from the control treatment were linked to their reduced photosynthetic capacity due to the high ASR severity. In uninfected plants, the ethylene concentration was not affected by Ni or Gl, while the ethylene concentration decreased in inoculated plants; this decrease was more pronounced in plants from the control treatment than in treated inoculated plants. In conclusion, this study sheds light on the role played by both Ni and Gl in ASR control from a physiological perspective. Soybean plants exposed to Ni and Gl were able to maintain high ethylene concentrations and photosynthetic capacity during the P. pachyrhizi infection process; as a result, these plants consumed less of their reserves than inoculated plants not treated with Ni or Gl.     

Reduction of Phakopsora pachyrhizi infection on soybean through host- and spray-induced gene silencing

Asian soybean rust (ASR), caused by the obligate fungal pathogen Phakopsora pachyrhizi, often leads to significant yield losses and can only be managed through fungicide applications currently. In the present study, eight urediniospore germination or appressorium formation induced P. pachyrhizi genes were investigated for their feasibility to suppress ASR through a bean pod mottle virus (BPMV)-based host-induced gene silencing (HIGS) strategy. Soybean plants expressing three of these modified BPMV vectors suppressed the expression of their corresponding target gene by 45%–80%, fungal biomass accumulation by 58%–80%, and significantly reduced ASR symptom development in soybean leaves after the plants were inoculated with P. pachyrhizi, demonstrating that HIGS can be used to manage ASR. In addition, when the in vitro synthesized double-stranded RNAs (dsRNAs) for three of the genes encoding an acetyl-CoA acyltransferase, a 40S ribosomal protein S16, and glycine cleavage system H protein were sprayed directly onto detached soybean leaves prior to P. pachyrhizi inoculation, they also resulted in an average of over 73% reduction of pustule numbers and 75% reduction in P. pachyrhizi biomass accumulation on the detached leaves compared to the controls. To the best of our knowledge, this is the first report of suppressing P. pachyrhizi infection in soybean through both HIGS and spray-induced gene silencing. It was demonstrated that either HIGS constructs targeting P. pachyrhizi genes or direct dsRNA spray application could be an effective strategy for reducing ASR development on soybean.     

Efficacy of Acibenzolar‐S‐Methyl and β‐Aminobutyric Acid for Control of Downy Mildew in Greenhouse Grown Basil and Peroxidase Activity in Response to Treatment with these Compounds

Downy mildew, caused by the oomycete pathogen Peronospora belbahrii, is a devastating foliar disease of basil in the United States and worldwide. Currently there are very few chemistries or organic choices registered to control this disease. In this study, two systemic acquired resistance (SAR) inducers, acibenzolar‐S‐methyl (ASM) and β‐aminobutyric acid (BABA), were evaluated for their in vitro effects on the pathogen, for their potential to control basil downy mildew in greenhouses, and for changes in peroxidase activity in basil plants treated with these two SAR inducers. No significant inhibition of sporangial germination was detected in water agar amended with ASM at concentrations lower than 100 mg/l or with BABA at concentrations lower than 500 mg/l. Efficacy of ASM and BABA in greenhouses varied depending on the rate, method and timing of application. The area under the disease progress curve (AUDPC) of disease severity was significantly reduced compared to the non‐treated control when ASM was sprayed (in all experiments) or drenched (in one out of two experiments) pre‐, or pre‐ + post‐inoculation at rates of 25–400 mg/l. Three weekly post‐inoculation sprays of ASM at the rate of 50 mg/l reduced AUDPC by 93.0 and 47.2% when started 3 and 7 days after inoculation (DAI), respectively. The AUDPC of disease severity was also significantly reduced when BABA was sprayed pre‐ + post‐inoculation at rates of 125–500 mg/l. According to the prediction using a log‐logistic function, 50% maximum disease protection was achieved at a concentration of 27.5 mg/l of ASM. Basil plants treated with these two SAR inducers and challenged with the pathogen showed significantly higher peroxidase activity than the non‐treated control at 8 DAI. Temporally, the highest activity of peroxidase was detected at 8 DAI, decreased at 15 DAI and waned further at 23 DAI.     

Rehydration of Sugar Beet Plants after Water Stress: Effect of Cytokinins

The possibility to improve the recovery of sugar beet plants after water stress by application of synthetic cytokinins N⁶-benzyladenine (BA) or N⁶-(m-hydroxybenzyl)adenosine (HBA) was tested. Relative water content (RWC), net photosynthetic rate (P_N), transpiration rate (E), stomatal conductance (g_s), chlorophyll (Chl) a and Chl b contents, and photosystem 2 efficiency characterized by variable to maximal fluorescence ratio (F_v/F_m) were measured in control plants, in water-stressed plants, and after rehydration (4, 8, 24, and 48 h). Water stress markedly decreased parameters of gas exchange, but they started to recover soon after irrigation. Application of BA or HBA to the substrate or sprayed on leaves only slightly stimulated recovery of P_N, E, and g_s in rehydrated plants, especially during the first phases of recovery. Chl contents decreased only under severe water stress and F_v/F_m ratio was not significantly affected by water stress applied. Positive effects of BA or HBA application on Chl content and F_v/F_m ratio were mostly not observed.     

Photosynthetic Characteristics of Two New Chlorophyll b-Less Rice Mutants

Two yellow rice mutants VG28-1 and VG30-5 were obtained during the tissue culture process from a rice plant (cv. Zhonghua No.11 japonica) with inserted maize Ds transposon element. Absorption spectra and pigment composition showed that two mutants had no chlorophyll (Chl) b and lower Chl a content in comparison to the wild type (WT). Net photosynthetic rate (P _N), total electron transport rate (J_F), photochemical quenching (q_p), quantum yield of PS2 dependent non-cyclic electron transport (_PS2), fraction of P_rate, and leaf area were lower but F_v/F_m and apparent quantum yield (AQY) remained at similar levels as in the WT plant. Xanthophyll cycle pool size (V+A+Z) on a Chl basis, and de-epoxidation state were enhanced in the mutants. The mutants had larger amounts of soluble protein and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO), especially the small subunit of RuBPCO, than WT. The characteristics of two rice mutants differed somewhat from the other common Chl b-less mutants originating from mutagenic agent treatments.     

Elevated CO2 enhances photosynthetic efficiency,ion uptake and antioxidant activity of Gynura bicolor DC. grown in a porous‐tube nutrient delivery system under simulated microgravity

It is well known that plants can grow under space conditions, however, perturbations of many biological phenomena have been highlighted due to the effect of altered gravity and its possible interaction with other factors (e.g., CO₂, ion radiation, etc. Our aim was to test whether elevated CO₂ could provide ‘protection’ to Gynura bicolor against the damaging effects of simulated microgravity (SM) on photosynthesis, ion uptake and antioxidant activity. As compared to G. bicolor grown in ambient CO₂ with no SM (ACO₂), growth and yield of the plants increased under elevated ambient CO₂ with no SM (ECO₂) and decreased under ACO₂+SM, whereas there was no significant effect on ECO₂+SM. Reductions in the content of Chl a, carotenoids and Chl a+b were 17.9%, 20.7% and 17.9% under ACO₂+SM, respectively, but under ECO₂ there was a significant effect on all photosynthetic pigments except Chl b, compared to ACO₂. Photosynthesis was improved under ECO₂ with SM and such an improvement was associated with improved water use efficiency and instantaneous carboxylation efficiency. Furthermore, SM caused a reduction in ion absorption rate, except for Ca²⁺, while ECO₂ increased the uptake rate. Finally, the activity of SOD, POD and the content of MDA and H₂O₂ were enhanced under SM treatments and were highest in ACO₂+SM. In contrast, T‐AOC activity and GSH content significantly declined in ACO₂+SM compared to other treatments. These results suggest that ACO₂ is not sufficient to counteract SM impact, but the increase is usually caused by improvement in CO₂ nutrition in ECO₂+SM in comparison with ACO₂+SM.     

Effects of water stress and high temperature on gas exchange and chlorophyll fluorescence in Triticum aestivum L.

Wheat plants grown in controlled growth chambers were exposed to drought stress (DS) and high temperature (HT) singly and in combination (DS+HT). The effects of these two stresses on net photosynthetic rate (P _N), stomatal conductance (g _s), intercellular CO₂ concentration (C _i), quantum efficiency of photosystem 2 (Φ_PS2), variable to maximum chlorophyll (Chl) fluorescence (F_v/F_m), photochemical (q_p) and non-photochemical (NPQ) Chl fluorescence, and yield were investigated. Grain yield was decreased by 21 % due to DS, while it was increased by 26 % due to HT. P _N, g _s, C _i, and Chl fluorescence were dramatically reduced to DS, HT, and their interaction, except NPQ which showed an increase due to HT.     

Photosynthetic responses of radish (<Emphasis Type="Italic">Raphanus sativus</Emphasis> var. <Emphasis Type="Italic">longipinnatus</Emphasis>) plants to infection by turnip mosaic virus

Plant growth, chlorophyll (Chl) content, photosynthetic gas exchange, ribulose-1,5-bisphosphate carboxylase (RuBPCO) enzyme activity, and Chl fluorescence in radish (Raphanus sativus var. longipinnatus) plants were examined after turnip mosaic virus (TuMV) infection. Plant fresh mass, dry mass, Chl content, net photosynthetic rate (P _N), transpiration rate (E), stomatal conductance (g _s), and RuBPCO activity were significantly lower in infected plants after 5 weeks of virus infection as compared to healthy plants. The 5-week virus infection did not induce significant differences in intercellular CO₂ concentration (C _i, photochemical efficiency of photosystem 2, PS2 (F_v/F_m), excitation capture efficiency of open PS2 reaction centres (F_v'/F_m'), effective quantum efficiency of photosystem 2 (ΔF/F_m'), and photochemical quenching (q_P), but non-photochemical quenching (q_N) and alternative electron sink (AES) were significantly enhanced. Thus the decreased plant biomass of TuMV-infected plants might be associated with the decreased photosynthetic activity mainly due to reduced RuBPCO activity.     

Responses of plant biomass,photosynthesis and lipid peroxidation to warming and precipitation change in two dominant species (Stipa grandis and Leymus chinensis) from North China Grasslands

Influential factors of global change affect plant carbon uptake and biomass simultaneously. Although the effects from warming and precipitation change have been extensive studied separately, the responses of plant biomass, photosynthesis, and lipid peroxidation to the interaction of these factors are still not fully understood. In this study, we examined the physiological responses of two dominant plant species from grasslands of northern China with different functional traits to combinations of five simulated warming patterns and five simulated precipitation patterns in environment‐controlled chambers. Our results showed that the biomass, net CO₂ assimilation rate (P_n), maximal efficiency of photosystem II photochemistry (F_v/F_m), and chlorophyll content (Chl) of Stipa grandis and Leymus chinensis were enhanced by moderate warming and plus precipitation, but they declined drastically with high temperature and drought. High temperature and drought also led to significant malondialdehyde (MDA) accumulation, which had a negative correlation with leaf biomass. The lower level of lipid peroxidation in leaves of S. grandis suggests that this species is better protected from oxidative damage under heat stress, drought stress and their interactive conditions than L. chinensis. Using the subordinate function values method, we found S. grandis to be more sensitive to climate change than L. chinensis and the gross biomass and root biomass of S. grandis and the leaf biomass of L. chinensis were most sensitive to climate change. Furthermore, the P_n of both S. grandis and L. chinensis had a significant linear relationship with F_v/F_m and Chl, indicating that carbon assimilation may be caused by nonstomatal limitations.     

Effects of rhizobia inoculation and nitrogen fertilization on photosynthetic physiology of soybean

Plant growth, contents of photosynthetic pigments, photosynthetic gas exchange, and chlorophyll (Chl) fluorescence in soybean [Glycine max (L.) Merr. cv. Heinong37] were investigated after it was inoculated with Sinorhizobium fredii USDA191 or treated with 5 mM (NH₄)₂SO₄ (N5) and 30 mM (NH₄)₂SO₄ (N30), respectively. In the plants following N5 fertilization, not only plant biomass, leaf area, and Chl content, but also net photosynthetic rate (P _N), stomatal conductance (g _s), carboxylation efficiency (CE), maximum photochemical efficiency (F_v/F_m) of photosystem 2 (PS2), and quantum yield of PS2 (Φ_PS2) were markedly improved as compared with the control plants. There were also positive effects on plant growth and plant photosynthesis after rhizobia inoculation, but the effects were much less than those of N5 fertilization. For N30 plants there were no significant positive effects on plant growth and photosynthetic capacity. Plant biomass, P _N, and g _s were similar to those of N-limited (control) plants. Φ_PS2 and photochemical quenching (q_P) were obviously declined while content of carotenoids and non-photochemical quenching (q_N) were significantly enhanced in N30 treated plants. This indicated that excess N supply may cause some negative effects on soybean plants.     

Effect of day/night temperature difference on chlorophyll content,photosynthesis and fluorescence parameters of tomato at fruit stage

In order to investigate the effect of day/night temperature difference (DIF) on photosynthetic characteristics of tomato plants (Solanum lycopersicum, cv. Jinguan 5) at fruit stage, an experiment was carried out in climate chambers. Five day/night temperature regimes (16/34, 19/31, 25/25, 31/19, and 34/16°C) with respective DIFs of -18, -12, 0, +12, and +18 were used and measured at mean daily temperature of 25°C. The results showed that chlorophyll (Chl) a, Chl b, net photosynthetic rate (P_N), stomatal conductance (g_s), maximum quantum yield of PSII photochemistry (F_v/F_m), effective quantum yield of PSII photochemistry (?_PSII), and photochemical quenching (qp) significantly increased under positive DIF, while they decreased with negative DIF. In contrast, the Chl a/b ratio and nonphotochemical quenching (NPQ) decreased under positive DIF, while increased with negative DIF. Chl a, Chl b, P_N, g_s, F_v/F_m, ?_PSII, and qp were larger under +12 DIF than those at +18 DIF, while Chl a/b and NPQ showed an opposite trend.     

Photosynthetic activity of poikilochlorophyllous desiccation tolerant plant <Emphasis Type="Italic">Reaumuria soongorica</Emphasis> during dehydration and re-hydration

Diurnal patterns of gas exchange and chlorophyll (Chl) fluorescence parameters of photosystem 2 (PS2) as well as Chl content were analyzed in Reaumuria soongorica (Pall.) Maxim., a perennial semi-shrub during dehydration and rehydration. The net photosynthetic rate (P _N), maximum photochemical efficiency of PS2 (variable to maximum fluorescence ratio, F_v/F_m), quantum efficiency of non-cyclic electron transport of PS2, and Chl content decreased, but non-photochemical quenching of fluorescence and carotenoid content increased in stems with the increasing of drought stress. 6 d after re-hydration, new leaves budded from stems. In the re-watered plants, the chloroplast function was restored and Chl a fluorescence returned to a similar level as in the control plants. This improved hydraulic adjustment in plant triggered a positive effect on ion flow in the tissues and increased shoot electrical admittance. Thus R. soongorica plants are able to sustain drought stress through leaf abscission and keep part of Chl content in stems.     

Genetics and mapping of adult plant rust resistance in soybean PI 587886 and PI 587880A

Two soybean accessions, PI 587886 and PI 587880A, previously identified as having resistance to Phakospora pachyrhizi Syd. (soybean rust, SBR) were used to create two populations (POP-1 and POP-2) segregating for SBR resistance. F₂-derived F₃ (F_2:3) families from each population were grown in a naturally SBR-infected field in Paraguay to determine inheritance and map resistance genes. Over 6,000 plants from 178 families in POP-1 and over 5,000 plants from 160 families in POP-2 were evaluated at R5 for lesion type: immune reaction (IR), reddish-brown (RB), or tan (TAN) colored lesions. Based on the lesion type present, each F_2:3 family was rated as resistant, segregating or susceptible and this classification was used to infer the F₂-phenotype and genotype. For both populations, the F₂ segregation ratios fit a 1:2:1 (resistant:segregating:susceptible) ratio expected for a single gene (P > 0.05). The RB lesions occurred almost exclusively in the heterozygous class, indicating incomplete dominance under the conditions of this study. Molecular markers flanking the locations of the known resistance genes were used to map the resistance gene in both populations to the Rpp1 locus. However, evaluation of PI 587886 and PI 587880A against eight P. pachyrhizi isolates indicated that the resistance allele in these two accessions was different from Rpp1. This test also demonstrated that these accessions were resistant to at least one P. pachyrhizi isolate collected in the southern US. This is the first report of using an adult plant field-screen with natural rust pressure to map SBR resistance.     

Prediction of the in planta Phakopsora pachyrhizi secretome and potential effector families

Asian soybean rust (ASR), caused by the obligate biotrophic fungus Phakopsora pachyrhizi, can cause losses greater than 80%. Despite its economic importance, there is no soybean cultivar with durable ASR resistance. In addition, the P. pachyrhizi genome is not yet available. However, the availability of other rust genomes, as well as the development of sample enrichment strategies and bioinformatics tools, has improved our knowledge of the ASR secretome and its potential effectors. In this context, we used a combination of laser capture microdissection (LCM), RNAseq and a bioinformatics pipeline to identify a total of 36 350 P. pachyrhizi contigs expressed in planta and a predicted secretome of 851 proteins. Some of the predicted secreted proteins had characteristics of candidate effectors: small size, cysteine rich, do not contain PFAM domains (except those associated with pathogenicity) and strongly expressed in planta. A comparative analysis of the predicted secreted proteins present in Pucciniales species identified new members of soybean rust and new Pucciniales‐ or P. pachyrhizi‐specific families (tribes). Members of some families were strongly up‐regulated during early infection, starting with initial infection through haustorium formation. Effector candidates selected from two of these families were able to suppress immunity in transient assays, and were localized in the plant cytoplasm and nuclei. These experiments support our bioinformatics predictions and show that these families contain members that have functions consistent with P. pachyrhizi effectors.     

Contrasting effects of high‐intensity photosynthetically active radiation on two bloom‐forming dinoflagellates

While light limitation can inhibit bloom formation in dinoflagellates, the potential for high‐intensity photosynthetically active radiation (PAR) to inhibit blooms by causing stress or damage has not been well‐studied. We measured the effects of high‐intensity PAR on the bloom‐forming dinoflagellates Alexandrium fundyense and Heterocapsa rotundata. Various physiological parameters (photosynthetic efficiency F_v/F_m, cell permeability, dimethylsulfoniopropionate [DMSP], cell volume, and chlorophyll‐a content) were measured before and after exposure to high‐intensity natural sunlight in short‐term light stress experiments. In addition, photosynthesis‐irradiance (P‐E) responses were compared for cells grown at different light levels to assess the capacity for photophysiological acclimation in each species. Experiments revealed distinct species‐specific responses to high PAR. While high light decreased F_v/F_m in both species, A. fundyense showed little additional evidence of light stress in short‐term experiments, although increased membrane permeability and intracellular DMSP indicated a response to handling. P‐E responses further indicated a high light‐adapted species with Chl‐a inversely proportional to growth irradiance and no evidence of photoinhibition; reduced maximum per‐cell photosynthesis rates suggest a trade‐off between photoprotection and C fixation in high light‐acclimated cells. Heterocapsa rotundata cells, in contrast, swelled in response to high light and sometimes lysed in short‐term experiments, releasing DMSP. P‐E responses confirmed a low light‐adapted species with high photosynthetic efficiencies associated with trade‐offs in the form of substantial photoinhibition and a lack of plasticity in Chl‐a content. These contrasting responses illustrate that high light constrains dinoflagellate community composition through species‐specific stress effects, with consequences for bloom formation and ecological interactions within the plankton.     

Chlorophyll fluorescence imaging of photosynthetic activity with the flash-lamp fluorescence imaging system

A flash-lamp chlorophyll (Chl) fluorescence imaging system (FL-FIS) is described that allows to screen and image the photosynthetic activity of several thousand leaf points (pixels) of intact leaves in a non-destructive way within a few seconds. This includes also the registration of several thousand leaf point images of the four natural fluorescence bands of plants in the blue (440 nm) and green (520 nm) regions as well as the red (near 690 nm) and far-red (near 740 nm) Chl fluorescence. The latest components of this Karlsruhe FL-FIS are presented as well as its advantage as compared to the classical single leaf point measurements where only the fluorescence information of one leaf point is sensed per each measurement. Moreover, using the conventional He-Ne-laser induced two-wavelengths Chl fluorometer LITWaF, we demonstrated that the photosynthetic activity of leaves can be determined measuring the Chl fluorescence decrease ratio, R_Fd (defined as Chl fluorescence decrease F_d from maximum to steady state fluorescence F_s:F_d/F_s), that is determined by the Chl fluorescence induction kinetics (Kautsky effect). The height of the values of the Chl fluorescence decrease ratio R_Fd is linearly correlated to the net photosynthetic CO₂ fixation rate P _N as is indicated here for sun and shade leaves of various trees that considerably differ in their P _N. Imaging the R_Fd-ratio of intact leaves permitted the detection of considerable gradients in photosynthetic capacity across the leaf area as well as the spatial heterogeneity and patchiness of photosynthetic quantum conversion within the control leaf and the stressed plants. The higher photosynthetic capacity of sun versus shade leaves was screened by Chl fluorescence imaging. Profile analysis of fluoresence signals (along a line across the leaf area) and histograms (the signal frequency distribution of the fluorescence information of all measured leaf pixels) of Chl fluorescence yield and Chl fluorescence ratios allow, with a high statistical significance, the quantification of the differences in photosynthetic activity between various areas of the leaf as well as between control leaves and water stressed leaves. The progressive uptake and transfer of the herbicide diuron via the petiole into the leaf of an intact plant and the concomitant loss of photosynthetic quantum conversion was followed with high precision by imaging the increase of the red Chl fluorescence F₆₉₀. Differences in the availability and absorption of soil nitrogen of crop plants can be documented via this flash-lamp fluorescence imaging technique by imaging the blue/red ratio image F₄₄₀/F₆₉₀, whereas differences in Chl content are detected by collecting images of the fluorescence ratio red/far-red, F₆₉₀/F₇₄₀.