Oxylipins and metabolites from pyroptotic cells act as promoters of tissue repair.

Pyroptosis is a lytic cell death mode that helps limit the spread of infections and is also linked to pathology in sterile inflammatory diseases and autoimmune diseases1-4. During pyroptosis, inflammasome activation and the engagement of caspase-1 lead to cell death, along with the maturation and secretion of the inflammatory cytokine interleukin-1ß (IL-1ß). The dominant effect of IL-1ß in promoting tissue inflammation has clouded the potential influence of other factors released from pyroptotic cells. Here, using a system in which macrophages are induced to undergo pyroptosis without IL-1ß or IL-1α release (denoted Pyro-1), we identify unexpected beneficial effects of the Pyro-1 secretome. First, we noted that the Pyro-1 supernatants upregulated gene signatures linked to migration, cellular proliferation and wound healing. Consistent with this gene signature, Pyro-1 supernatants boosted migration of primary fibroblasts and macrophages, and promoted faster wound closure in vitro and improved tissue repair in vivo. In mechanistic studies, lipidomics and metabolomics of the Pyro-1 supernatants identified the presence of both oxylipins and metabolites, linking them to pro-wound-healing effects. Focusing specifically on the oxylipin prostaglandin E2 (PGE2), we find that its synthesis is induced de novo during pyroptosis, downstream of caspase-1 activation and cyclooxygenase-2 activity; further, PGE2 synthesis occurs late in pyroptosis, with its release dependent on gasdermin D pores opened during pyroptosis. As for the pyroptotic metabolites, they link to immune cell infiltration into the wounds, and polarization to CD301+ macrophages. Collectively, these data advance the concept that the pyroptotic secretome possesses oxylipins and metabolites with tissue repair properties that may be harnessed therapeutically.

Local Action of Moderate Heating and Illumination Induces Electrical Signals, Suppresses Photosynthetic Light Reactions, and Increases Drought Tolerance in Wheat Plants.

Local actions of stressors induce electrical signals (ESs), influencing photosynthetic processes and probably increasing tolerance to adverse factors in higher plants. However, the participation of well-known depolarization ESs (action potentials and variation potentials) in these responses seems to be rare under natural conditions, particularly in the case of variation potentials, which are induced by extreme stressors (e.g., burning). Earlier, we showed that the local action of moderate heating and illumination can induce low-amplitude hyperpolarization ESs influencing photosynthetic light reactions in wheat plants cultivated in a vegetation room. In the current work, we analyzed ESs and changes in photosynthetic light reactions and drought tolerance that were induced by a combination of moderate heating and illumination in wheat plants cultivated under open-ground conditions. It was shown that the local heating and illumination induced low-amplitude ESs, and the type of signal (depolarization or hyperpolarization) was dependent on distance from the irritated zone and wheat age. Induction of depolarization ESs was not accompanied by photosynthetic changes in plants under favorable conditions or under weak drought. In contrast, the changes were observed after induction of these signals under moderate drought. Increasing drought tolerance was also observed in the last case. Thus, low-amplitude ESs can participate in photosynthetic regulation and increase tolerance to drought in plants cultivated under open-ground conditions.

RGB Imaging as a Tool for Remote Sensing of Characteristics of Terrestrial Plants: A Review.

Approaches for remote sensing can be used to estimate the influence of changes in environmental conditions on terrestrial plants, providing timely protection of their growth, development, and productivity. Different optical methods, including the informative multispectral and hyperspectral imaging of reflected light, can be used for plant remote sensing; however, multispectral and hyperspectral cameras are technically complex and have a high cost. RGB imaging based on the analysis of color images of plants is definitely simpler and more accessible, but using this tool for remote sensing plant characteristics under changeable environmental conditions requires the development of methods to increase its informativity. Our review focused on using RGB imaging for remote sensing the characteristics of terrestrial plants. In this review, we considered different color models, methods of exclusion of background in color images of plant canopies, and various color indices and their relations to characteristics of plants, using regression models, texture analysis, and machine learning for the estimation of these characteristics based on color images, and some approaches to provide transformation of simple color images to hyperspectral and multispectral images. As a whole, our review shows that RGB imaging can be an effective tool for estimating plant characteristics; however, further development of methods to analyze color images of plants is necessary.

Changes in Activity of the Plasma Membrane H+-ATPase as a Link Between Formation of Electrical Signals and Induction of Photosynthetic Responses in Higher Plants.

Action of numerous adverse environmental factors on higher plants is spatially-heterogenous; it means that induction of a systemic adaptive response requires generation and transmission of the stress signals. Electrical signals (ESs) induced by local action of stressors include action potential, variation potential, and system potential and they participate in formation of fast physiological changes at the level of a whole plant, including photosynthetic responses. Generation of these ESs is accompanied by the changes in activity of H+-ATPase, which is the main system of electrogenic proton transport across the plasma membrane. Literature data show that the changes in H+-ATPase activity and related changes in intra- and extracellular pH play a key role in the ES-induced inactivation of photosynthesis in non-irritated parts of plants. This inactivation is caused by both suppression of CO2 influx into mesophyll cells in leaves, which can be induced by the apoplast alkalization and, probably, cytoplasm acidification, and direct influence of acidification of stroma and lumen of chloroplasts on light and, probably, dark photosynthetic reactions. The ES-induced inactivation of photosynthesis results in the increasing tolerance of photosynthetic machinery to the action of adverse factors and probability of the plant survival.

Single-cell atlas of healthy human blood unveils age-related loss of NKG2C+GZMB-CD8+ memory T cells and accumulation of type 2 memory T cells.

Extensive, large-scale single-cell profiling of healthy human blood at different ages is one of the critical pending tasks required to establish a framework for the systematic understanding of human aging. Here, using single-cell RNA/T cell receptor (TCR)/BCR-seq with protein feature barcoding, we profiled 317 samples from 166 healthy individuals aged 25-85 years old. From this, we generated a dataset from ∼2 million cells that described 55 subpopulations of blood immune cells. Twelve subpopulations changed with age, including the accumulation of GZMK+CD8+ T cells and HLA-DR+CD4+ T cells. In contrast to other T cell memory subsets, transcriptionally distinct NKG2C+GZMB-CD8+ T cells counterintuitively decreased with age. Furthermore, we found a concerted age-associated increase in type 2/interleukin (IL)4-expressing memory subpopulations across CD4+ and CD8+ T cell compartments (CCR4+CD8+ Tcm and Th2 CD4+ Tmem), suggesting a systematic functional shift in immune homeostasis with age. Our work provides novel insights into healthy human aging and a comprehensive annotated resource.

Development of Modified Farquhar-von Caemmerer-Berry Model Describing Photodamage of Photosynthetic Electron Transport in C3 Plants under Different Temperatures.

Photodamage of photosynthetic electron transport is a key mechanism of disruption of photosynthesis in plants under action of stressors. This means that investigation of photodamage is an important task for basic and applied investigations. However, its complex mechanisms restrict using experimental methods of investigation for this process; the development of mathematical models of photodamage and model-based analysis can be used for overcoming these restrictions. In the current work, we developed the modified Farquhar-von Caemmerer-Berry model which describes photodamage of photosynthetic electron transport in C3 plants. This model was parameterized on the basis of experimental results (using an example of pea plants). Analysis of the model showed that combined inactivation of linear electron flow and Rubisco could induce both increasing and decreasing photodamage at different magnitudes of inactivation of these processes. Simulation of photodamage under different temperatures and light intensities showed that simulated temperature dependences could be multi-phase; particularly, paradoxical increases in the thermal tolerance of photosynthetic electron transport could be observed under high temperatures (37-42 °C). Finally, it was shown that changes in temperature optimums of linear electron flow and Rubisco could modify temperature dependences of the final activity of photosynthetic electron transport under photodamage induction; however, these changes mainly stimulated its photodamage. Thus, our work provides a new theoretical tool for investigation of photodamage of photosynthetic processes in C3 plants and shows that this photodamage can be intricately dependent on parameters of changes in activities of linear electron flow and Rubisco including changes induced by temperature.

Local Action of Increased Pressure Induces Hyperpolarization Electrical Signals and Influences Photosynthetic Light Reactions in Wheat Plants.

Long-distance electrical signals caused by the local action of stressors influence numerous physiological processes in plants including photosynthesis and increase their tolerance to the action of adverse factors. Depolarization electrical signals were mainly investigated; however, we earlier showed that hyperpolarization electrical signals (HESs) can be caused by moderate stressors (e.g., local moderate heating) and induce photosynthetic inactivation. We hypothesized that HESs are related to stressor-induced increases in the hydrostatic pressure in the zone of action of the stressor and following the propagation of a hydraulic wave. In the current work, we tested this hypothesis through the direct investigation of electrical signals induced by the local action of artificially increased pressure and an analysis of the subsequent photosynthetic changes in the nonirritated parts of plants. The electrical signals and parameters of photosynthetic light reactions were investigated in wheat plants. The local action of the increased pressure was induced by the action of weights on the wheat leaf. Extracellular electrodes were used for electrical signal measurements. Pulse-amplitude-modulation fluorescent imaging was used for measurements of the quantum yield of photosystem II and nonphotochemical quenching of chlorophyll fluorescence in wheat leaves. It was shown that the local action of pressure on wheat leaf induced electrical signals near the irritated zone: HESs were caused by low pressure (10 kPa) and depolarization signals were induced by high pressure (100 kPa). The local action of moderate pressure (50 kPa) induced weak electrical signals near the irritated zone; however, HESs were observed with increasing distance from this zone. It was also shown that the local action of this moderate pressure induced the photosynthetic inactivation (decreasing the quantum yield of photosystem II and increasing the nonphotochemical quenching of chlorophyll fluorescence) in the nonirritated parts of the wheat leaves. Thus, our results show that the local action of the increased pressure and, probably, subsequent propagation of the hydraulic wave induce electrical signals (including HESs) and photosynthetic inactivation in nonirritated parts of plants that are similar to ones caused by the local action of moderate stressors (e.g., moderate heating). This means that both HESs and depolarization electrical signals can have a hydraulic mechanism of propagation.

Hyperpolarization electrical signals induced by local action of moderate heating influence photosynthetic light reactions in wheat plants.

Local action of stressors induces fast changes in physiological processes in intact parts of plants including photosynthetic inactivation. This response is mediated by generation and propagation of depolarization electrical signals (action potentials and variation potentials) and participates in increasing plant tolerance to action of adverse factors. Earlier, we showed that a local action of physiological stimuli (moderate heating and blue light), which can be observed under environmental conditions, induces hyperpolarization electrical signals (system potentials) in wheat plants. It potentially means that these signals can play a key role in induction of fast physiological changes under the local action of environmental stressors. The current work was devoted to investigation of influence of hyperpolarization electrical signals induced by the local action of the moderate heating and blue light on parameters of photosynthetic light reactions. A quantum yield of photosystem II (ФPSII) and a non-photochemical quenching of chlorophyll fluorescence (NPQ) in wheat plants were investigated. It was shown that combination of the moderate heating (40°C) and blue light (540 µmol m-2s-1) decreased ФPSII and increased NPQ; these changes were observed in 3-5 cm from border of the irritated zone and dependent on intensity of actinic light. The moderate soil drought (7 days) increased magnitude of photosynthetic changes and shifted their localization which were observed on 5-7 cm from the irritated zone; in contrast, the strong soil drought (14 days) suppressed these changes. The local moderate heating decreased ФPSII and increased NPQ without action of the blue light; in contrast, the local blue light action without heating weakly influenced these parameters. It meant that just local heating was mechanism of induction of the photosynthetic changes. Finally, propagation of hyperpolarization electrical signals (system potentials) was necessary for decreasing ФPSII and increasing NPQ. Thus, our results show that hyperpolarization electrical signals induced by the local action of the moderate heating inactivates photosynthetic light reactions; this response is similar with photosynthetic changes induced by depolarization electrical signals. The soil drought and actinic light intensity can influence parameters of these photosynthetic changes.

Effect of Duration of LED Lighting on Growth, Photosynthesis and Respiration in Lettuce.

Parameters of illumination including the spectra, intensity, and photoperiod play an important role in the cultivation of plants under greenhouse conditions, especially for vegetables such as lettuce. We previously showed that illumination by a combination of red, blue, and white LEDs with a high red light intensity, was optimal for lettuce cultivation; however, the effect of the photoperiod on lettuce cultivation was not investigated. In the current work, we investigated the influence of photoperiod on production (total biomass and dry weight) and parameters of photosynthesis, respiration rate, and relative chlorophyll content in lettuce plants. A 16 h (light):8 h (dark) illumination regime was used as the control. In this work, we investigated the effect of photoperiod on total biomass and dry weight production in lettuce plants as well as on photosynthesis, respiration rate and chlorophyll content. A lighting regime 16:8 h (light:dark) was used as control. A shorter photoperiod (8 h) decreased total biomass and dry weight in lettuce, and this effect was related to the suppression of the linear electron flow caused by the decreasing content of chlorophylls and, therefore, light absorption. A longer photoperiod (24 h) increased the total biomass and dry weight, nevertheless an increase in photosynthetic processes, light absorption by leaves and chlorophyll content was not recorded, nor were differences in respiration rate, thus indicating that changes in photosynthesis and respiration are not necessary conditions for stimulating plant production. A simple model to predict plant production was also developed to address the question of whether increasing the duration of illumination stimulates plant production without inducing changes in photosynthesis and respiration. Our results indicate that increasing the duration of illumination can stimulate dry weight accumulation and that this effect can also be induced using the equal total light integrals for day (i.e., this stimulation can be also caused by increasing the light period while decreasing light intensity). Increasing the duration of illumination is therefore an effective approach to stimulating lettuce production under artificial lighting.

Development of Two-Dimensional Model of Photosynthesis in Plant Leaves and Analysis of Induction of Spatial Heterogeneity of CO2 Assimilation Rate under Action of Excess Light and Drought.

Photosynthesis is a key process in plants that can be strongly affected by the actions of environmental stressors. The stressor-induced photosynthetic responses are based on numerous and interacted processes that can restrict their experimental investigation. The development of mathematical models of photosynthetic processes is an important way of investigating these responses. Our work was devoted to the development of a two-dimensional model of photosynthesis in plant leaves that was based on the Farquhar-von Caemmerer-Berry model of CO2 assimilation and descriptions of other processes including the stomatal and transmembrane CO2 fluxes, lateral CO2 and HCO3- fluxes, transmembrane and lateral transport of H+ and K+, interaction of these ions with buffers in the apoplast and cytoplasm, light-dependent regulation of H+-ATPase in the plasma membrane, etc. Verification of the model showed that the simulated light dependences of the CO2 assimilation rate were similar to the experimental ones and dependences of the CO2 assimilation rate of an average leaf CO2 conductance were also similar to the experimental dependences. An analysis of the model showed that a spatial heterogeneity of the CO2 assimilation rate on a leaf surface should be stimulated under an increase in light intensity and a decrease in the stomatal CO2 conductance or quantity of the open stomata; this prediction was supported by the experimental verification. Results of the work can be the basis of the development of new methods of the remote sensing of the influence of abiotic stressors (at least, excess light and drought) on plants.

Simulated Analysis of Influence of Changes in H+-ATPase Activity and Membrane CO2 Conductance on Parameters of Photosynthetic Assimilation in Leaves.

Photosynthesis is an important process in plants which influences their development and productivity. Many factors can control the efficiency of photosynthesis, including CO2 conductance of leaf mesophyll, which affects the CO2 availability for Rubisco. It is known that electrical stress signals can decrease this conductance, and the response is probably caused by inactivation of H+-ATPase in the plasma membrane. In the current work, we analyzed the influence of both CO2 conductance in the plasma membrane, and chloroplast envelopes and H+-ATPase activity on photosynthetic CO2 assimilation, using a two-dimensional mathematical model of photosynthesis in leaves. The model included a description of assimilation on the basis of the Farquhar-von Caemmerer-Berry model, ion transport through the plasma membrane, diffusion of CO2 in the apoplast, and transport of CO2 through the plasma membrane and chloroplast envelope. The model showed that the photosynthetic CO2 assimilation rate was mainly dependent on the plasma membrane and chloroplast envelope conductance; direct influence of the H+-ATPase activity (through changes in pH and CO2/HCO3- concentration ratio) on this rate was weak. In contrast, both changes in CO2 conductance of the plasma membrane and chloroplast envelopes and changes in the H+-ATPase activity influenced spatial heterogeneity of the CO2 assimilation on the leaf surface in the simulated two-dimensional system. These effects were also observed under simultaneous changes in the CO2 conductance of the plasma membrane and H+-ATPase activity. Qualitatively similar influence of changes in the CO2 conductance of the plasma membrane and chloroplast envelopes, and changes in the H+-ATPase activity on photosynthesis were shown for two different densities of stomata in the simulated leaf; however, lowering the density of stomata decreased the assimilation rate and increased the heterogeneity of assimilation. The results of the model analysis clarify the potential influence of H+-ATPase inactivation on photosynthesis, and can be the basis for development of new methods for remote sensing of the influence of electrical signals.

Modified Photochemical Reflectance Indices as New Tool for Revealing Influence of Drought and Heat on Pea and Wheat Plants.

In environmental conditions, plants can be affected by the action of numerous abiotic stressors. These stressors can induce both damage of physiological processes and adaptive changes including signaling-based changes. Development of optical methods of revealing influence of stressors on plants is an important task for plant investigations. The photochemical reflectance index (PRI) based on plant reflectance at 531 nm (measuring wavelength) and 570 nm (reference wavelength) can be effective tool of revealing plant stress changes (mainly, photosynthetic changes); however, its efficiency is strongly varied at different conditions. Earlier, we proposed series of modified PRIs with moderate shifts of the measuring wavelength and showed that these indices can be effective for revealing photosynthetic changes under fluctuations in light intensity. The current work was devoted to the analysis of sensitivity of these modified PRIs to action of drought and short-term heat stress. Investigation of spatially-fixed leaves of pea plants showed that the modified PRI with the shorter measuring wavelength (515 nm) was increased under response of drought and heat; by contrast, the modified PRI with the longer wavelength (555 nm) was decreased under response to these stressors. Changes of investigated indices could be related to parameters of photosynthetic light reactions; however, these relations were stronger for the modified PRI with the 555 nm measuring wavelength. Investigation of canopy of pea (vegetation room) and wheat (vegetation room and open-ground) supported these results. Thus, moderate changes in the measuring wavelengths of PRI can strongly modify the efficiency of their use for the estimation of plant physiological changes (mainly photosynthetic changes) under action of stressors. It is probable that the modified PRI with the 555 nm measuring wavelength (or similar indices) can be an effective tool for revealing photosynthetic changes induced by stressors.

Influence of Burning-Induced Electrical Signals on Photosynthesis in Pea Can Be Modified by Soil Water Shortage.

Local damage to plants can induce fast systemic physiological changes through generation and propagation of electrical signals. It is known that electrical signals influence numerous physiological processes including photosynthesis; an increased plant tolerance to actions of stressors is a result of these changes. It is probable that parameters of electrical signals and fast physiological changes induced by these signals can be modified by the long-term actions of stressors; however, this question has been little investigated. Our work was devoted to the investigation of the parameters of burning-induced electrical signals and their influence on photosynthesis under soil water shortage in pea seedlings. We showed that soil water shortage decreased the amplitudes of the burning-induced depolarization signals (variation potential) and the magnitudes of photosynthetic inactivation (decreasing photosynthetic CO2 assimilation and linear electron flow and increasing non-photochemical quenching of the chlorophyll fluorescence and cyclic electron flow around photosystem I) caused by these signals. Moreover, burning-induced hyperpolarization signals (maybe, system potentials) and increased photosynthetic CO2 assimilation could be observed under strong water shortage. It was shown that the electrical signal-induced increase of the leaf stomatal conductance was a potential mechanism for the burning-induced activation of photosynthetic CO2 assimilation under strong water shortage; this mechanism was not crucial for photosynthetic response under control conditions or weak water shortage. Thus, our results show that soil water shortage can strongly modify damage-induced electrical signals and fast physiological responses induced by these signals.

Ratio of Intensities of Blue and Red Light at Cultivation Influences Photosynthetic Light Reactions, Respiration, Growth, and Reflectance Indices in Lettuce.

LED illumination can have a narrow spectral band; its intensity and time regime are regulated within a wide range. These characteristics are the potential basis for the use of a combination of LEDs for plant cultivation because light is the energy source that is used by plants as well as the regulator of photosynthesis, and the regulator of other physiological processes (e.g., plant development), and can cause plant damage under certain stress conditions. As a result, analyzing the influence of light spectra on physiological and growth characteristics during cultivation of different plant species is an important problem. In the present work, we investigated the influence of two variants of LED illumination (red light at an increased intensity, the "red" variant, and blue light at an increased intensity, the "blue" variant) on the parameters of photosynthetic dark and light reactions, respiration rate, leaf reflectance indices, and biomass, among other factors in lettuce (Lactuca sativa L.). The same light intensity (about 180 µmol m-2s-1) was used in both variants. It was shown that the blue illumination variant increased the dark respiration rate (35-130%) and cyclic electron flow around photosystem I (18-26% at the maximal intensity of the actinic light) in comparison to the red variant; the effects were dependent on the duration of cultivation. In contrast, the blue variant decreased the rate of the photosynthetic linear electron flow (13-26%) and various plant growth parameters, such as final biomass (about 40%). Some reflectance indices (e.g., the Zarco-Tejada and Miller Index, an index that is related to the core sizes and light-harvesting complex of photosystem I), were also strongly dependent on the illumination variant. Thus, our results show that the red illumination variant contributes a great deal to lettuce growth; in contrast, the blue variant contributes to stress changes, including the activation of cyclic electron flow around photosystem I.

Effect of Photoconversion Coatings for Greenhouses on Electrical Signal-Induced Resistance to Heat Stress of Tomato Plants.

The use of photoconversion coatings is a promising approach to improving the quality of light when growing plants in greenhouses in low light conditions. In this work, we studied the effect of fluoropolymer coatings, which produce photoconversion of UV-A radiation and violet light into blue and red light, on the growth and resistance to heat stress of tomato plants (Solanum lycopersicum L.). The stimulating effect of the spectrum obtained as a result of photoconversion on plant growth and the activity of the photosynthesis process are shown. At the same time, the ability to withstand heat stress is reduced in plants grown under a photoconversion coating. Stress electrical signals, which normally increase resistance, in such plants have a much weaker protective effect on the photosynthetic apparatus. The observed effects are apparently explained by a decrease in the concentration of H2O2 in plants grown using photoconversion technologies, which leads to a shift in the development program towards increased productivity to the detriment of the protective function. Thus, when using photoconversion technologies in agricultural practice, it is necessary to pay increased attention to maintaining stable conditions during plant cultivation.

Effect of extremely low-frequency magnetic fields on light-induced electric reactions in wheat.

Magnetic field oscillations resulting from atmospheric events could have an effect on growth and development of plants and on the responsive reactions of plants to other environmental factors. In the current work, extremely low-frequency magnetic field (14.3 Hz) was shown to modulate light-induced electric reactions of wheat (Triticum aestivum L.). Blue light-induced electric reaction in wheat leaf comprises depolarization and two waves of hyperpolarization resulting in an increase of the potential to a higher level compared to the dark one. Fluorescent and inhibitory analysis demonstrate a key role of calcium ions and calcium-dependent H+-ATPase of the plasma membrane in the development of the reaction. Activation of H+-ATPase by the increased calcium influx is suggested as a mechanism of the influence of magnetic field on light-induced electric reaction.

Analysis of chlorophyll fluorescence parameters as predictors of biomass accumulation and tolerance to heat and drought stress of wheat (Triticum aestivum) plants.

Agricultural technologies aimed at increasing yields require the development of highly productive and stress-tolerant cultivars. Phenotyping can significantly accelerate breeding; however, no reliable markers have been identified to select the most promising cultivars at an early stage. In this work, we determined the light-induced dynamic of chlorophyll fluorescence (ChlF) parameters in young seedlings of 10 wheat (Triticum aestivum L.) cultivars and evaluated potency of these parameters as predictors of biomass accumulation and stress tolerance. Dry matter accumulation positively correlated with the effective quantum efficiency of photosystem II (Φ PSIIef ) and negatively correlated with the half-time of Φ PSIIef reaching (t 1/2 (Φ PSIIef )). There was a highly significant correlation between t 1/2 (Φ PSIIef ) and dry matter accumulation with increasing prediction period. Short-term heating and drought caused an inhibition of biomass accumulation and photosynthetic activity depending on the stressor intensity. The positive correlation between the Φ PSII dark level (Φ PSIId ) in young seedlings and tolerance to a rapidly increasing short-term stressor (heating) was shown. In the case of a long-term stressor (drought), we revealed a strong negative relationship between tolerance and the level of non-photochemical fluorescence quenching (NPQ). In general, the results show the potency of the ChlF parameters of young seedlings as predictors of biomass accumulation and stress tolerance.

Pre-Symptomatic Detection of Viral Infection in Tobacco Leaves Using PAM Fluorometry.

Chlorophyll fluorescence imaging was used to study potato virus X (PVX) infection of Nicotiana benthamiana. Infection-induced changes in chlorophyll fluorescence parameters (quantum yield of photosystem II photochemistry (ΦPSII) and non-photochemical fluorescence quenching (NPQ)) in the non-inoculated leaf were recorded and compared with the spatial distribution of the virus detected by the fluorescence of GFP associated with the virus. We determined infection-related changes at different points of the light-induced chlorophyll fluorescence kinetics and at different days after inoculation. A slight change in the light-adapted steady-state values of ΦPSII and NPQ was observed in the infected area of the non-inoculated leaf. In contrast to the steady-state parameters, the dynamics of ΦPSII and NPQ caused by the dark-light transition in healthy and infected areas differed significantly starting from the second day after the detection of the virus in a non-inoculated leaf. The coefficients of correlation between chlorophyll fluorescence parameters and virus localization were 0.67 for ΦPSII and 0.76 for NPQ. In general, the results demonstrate the possibility of reliable pre-symptomatic detection of the spread of a viral infection using chlorophyll fluorescence imaging.