Chlorophyll Fluorescence Kinetics Research Articles

Fast chlorophyll fluorescence rise kinetics as a high-throughput diagnostic tool for evaluating the capacity of 2-amino-3-methylhexanoic acid at inducing plant resistance against high temperature

Plant resistant induction is considered as a promising strategy for protecting crops against extreme high temperature (HT). However, a high-throughput method to accurately estimate the capacity of plant resistance inducers (PRIs) for HT resistance has not been developed. Here, we present a simple approach using fast chlorophyll fluorescence kinetics in Arabidopsis leaf discs to assess PRI efficacy in inducing HT resistance. Both 2-amino-3-methylhexanoic acid (AMHA) and salicylic acid (SA) significantly alleviated the temperature-dependent increase in the K-peak of the OJIP curve and variations in amplitude of heat-responsive JIP-test parameters within the elevated-temperature range of 25 to 42℃. The PIABS (performance index on absorption basis) and WK (relative variable fluorescence at the K-step to the amplitude FJ - FO) as two classical heat-responsive characteristic parameters were used to produce a novel hypersensitive parameter HT sensitivity indicator, PIABS/WK (named Hs). Based on the correlation of logHs with elevated temperatures, a model for quantifying the capacity of HT-resistance induction (called Ci) by AMHA or SA was established. A three-grade classification according to the Ci value was proposed as low (0<Ci ≤ 1℃), moderate (1℃ <Ci ≤ 2℃), and high resistance (Ci > 2℃). AMHA at 1µM and SA at 100µM had Ci values of 2.49℃ and 4.09℃ in Arabidopsis plants, respectively, associated with their high level of HT resistance induction. Additionally, the EC50 derived from the relative stimulation ratio (Kc) was also introduced as a quantitative index for measuring the ability of AMHA and SA to induce HT resistance. The EC50 value of AMHA is about 0.1µM in Arabidopsis and 0.35µM in tomato, being much lower than that of SA (approximately 63µM in Arabidopsis). Thus, AMHA is a more potent plant inducer than SA. The model was validated through additional experimental evidence, demonstrating its reliability and applicability. This study provides an expeditious high-throughput method for screening promising PRI candidates.

Metabolomics combined with proteomics reveals phytotoxic effects of norfloxacin under drought stress on Oryza sativa

In recent decades, plants enduring abiotic stresses such as drought and chemical stresses. Currently, the mechanism of combined antibiotic and drought stress response and its impact on crop growth and food security remains poorly understood. Here, the mechanism of stress responses under the exposure of norfloxacin (NF) and drought (D) individually and in combination (DNF) were explored on rice (Oryza sativa) cultivar Hanyou73 through proteomics and metabolomic analysis. All treatments adversely affected chlorophyll fluorescence kinetics, antioxidant enzyme activities, rice grain composition and yield. The results showed that in DNF the antibiotic was accumulated 627% more than NF treatment in rice grains while in leaves there was no significant difference under both treatments. The proteomic revealed that differentially expressed identified proteins were involved in carbohydrate metabolism, amino acid metabolism, photosynthesis and mRNA binding. However, the metabolomics results showed that the abundance of metabolites related to RNA biosynthesis and amino acid metabolism were more affected. The disruptions caused in rice plant under DNF treatment become more severe, this makes it more susceptible than individual D and NF treatment. These findings improve our knowledge about the response of rice plant to cope with antibiotic contamination alone and in combination with drought.

Optimized nitrogen application ameliorates the photosynthetic performance and yield potential in peanuts as revealed by OJIP chlorophyll fluorescence kinetics

BackgroundNitrogen (N) is a crucial element for increasing photosynthesis and crop yields. The study aims to evaluate the photosynthetic regulation and yield formation mechanisms of different nodulating peanut varieties with N fertilizer application.MethodThe present work explored the effect of N fertilizer application rates (N0, N45, N105, and N165) on the photosynthetic characteristics, chlorophyll fluorescence characteristics, dry matter, N accumulation, and yield of four peanut varieties.ResultsThe results showed that N application increased the photosynthetic capacity, dry matter, N accumulation, and yield of peanuts. The measurement of chlorophyll a fluorescence revealed that the K-phase, J-phase, and I-phase from the OJIP curve decreased under N105 treatment compared with N0, and WOI, ET0/CSM, RE0/CSM, ET0/RC, RE0/RC, φPo, φEo, φRo, and Ψ0 increased, whereas VJ, VI, WK, ABS/RC, TR0/RC, DI0/RC, and φDo decreased. Meanwhile, the photosystem activity and electron transfer efficiency of nodulating peanut varieties decreased with an increase in N (N165). However, the photosynthetic capacity and yield of the non-nodulating peanut variety, which highly depended on N fertilizer, increased with an increase in N.ConclusionOptimized N application (N105) increased the activity of the photosystem II (PSII) reaction center, improved the electron and energy transfer performance in the photosynthetic electron transport chain, and reduced the energy dissipation of leaves in nodulating peanut varieties, which is conducive to improving the yield. Nevertheless, high N (N165) had a positive effect on the photosystem and yield of non-nodulating peanut. The results provide highly valuable guidance for optimizing peanut N management and cultivation measures.

Addressing the altitudinal and geographical gradient in European beech via photosynthetic parameters: a case study on Calabrian beech transplanted to Denmark

European beech (Fagus sylvatica L.) is becoming one of the go-to species in reconstructing declining conifer stands in Europe under climate change. Assisted migration may be considered when looking for suitable beech seedlings. Knowledge about the photosynthetic performance of beech seedlings is fundamental to understanding an essential part of their growth and survival potential in different planting conditions. We investigated the within-provenance variation in photosynthetic performance driven by altitude in contrast to inter-provenance variation given by geographical distance. The experiment was conducted on seedlings replanted in a Danish common garden comprising a cluster of provenances with various altitudinal subgroups transplanted from the Calabria region (Italy) and two local Danish provenances. Provenance and within-provenance variation in chlorophyll fluorescence (ChlF) kinetics, gas exchange (GE), flushing, and senescence were assessed. ChlF measurements revealed within-provenance differences based on altitude of origin and could distinguish between the two Danish provenances. In contrast, GE parameters detected variation in the geographical distance among Italian provenances. High-elevation subgroups of Italian provenances showed the best leaf-level photosynthetic performance in Danish weather conditions with high precipitation levels. Altitude of origin can be a significant source of within-provenance variation. We demonstrated that assessing this variation in young trees may be instrumental in maximizing the potential of provenance variation across diverse planting sites.

Erratum to: Effects of Chlormequat Chloride and Paclobutrazol on the Growth and Chlorophyll Fluorescence Kinetics of Daphne genkwa

Erratum to: Effects of Chlormequat Chloride and Paclobutrazol on the Growth and Chlorophyll Fluorescence Kinetics of Daphne genkwa

Ultrastructure, CO2 Assimilation and Chlorophyll Fluorescence Kinetics in Photosynthesizing Glycine max Callus and Leaf Mesophyll Tissues

The ultrastructural and functional features of photosynthesizing callus cells are poorly known. Electron microscopy studies on green, compact Glycine max calluses have shown that they are composed of photosynthesizing cells characterized by clear ultrastructural signs of senescence. Studies on chlorophyll fluorescence and CO2 assimilation kinetics have shown that such cells were still able to maintain photosynthesis but could not compensate for the respiratory CO2 uptake. Having a one-step CO2 assimilation kinetics, photosynthesis in calluses differed from photosynthesis in leaves, which had a two-step CO2 assimilation kinetics. In contrast to leaves, the fluorescence induction curves in G. max calluses strongly differed in shape depending on the color of actinic light (red or blue). Red (in contrast to blue) light excitation did not lead to CO2 assimilation in the calluses, thus suggesting anoxygenic photosynthesis in this case. In particular, the data obtained indicate that the actinic light spectrum should be considered when cultivating calluses for micropropagation of plants and for callus tissue research.

Transcriptome and photosynthetic analyses provide new insight into the molecular mechanisms underlying heat stress tolerance in Rhododendron × pulchrum Sweet.

Rhododendron species provide excellent ornamental use worldwide, yet heat stress (HS) is one of the major threats to their cultivation. However, the intricate mechanisms underlying the photochemical and transcriptional regulations associated with the heat stress response in Rhododendron remain relatively unexplored. In this study, the analyses of morphological characteristics and chlorophyll fluorescence (ChlF) kinetics showed that HS (40°C/35°C) had a notable impact on both the donor's and acceptor's sides of photosystem II (PSII), resulting in reduced PSII activity and electron transfer capacity. The gradual recovery of plants observed following a 5-day period of culture under normal conditions indicates the reversible nature of the HS impact on Rhododendron × pulchrum. Analysis of transcriptome data unveiled noteworthy trends: four genes associated with photosynthesis-antenna protein synthesis (LHCb1, LHCb2 and LHCb3) and the antioxidant system (glutamate-cysteine ligase) experienced significant down-regulation in the leaves of R. × pulchrum during HS. Conversely, aseorbate peroxidase and glutathione S-transferase TAU 8 demonstrated an up-regulated pattern. Furthermore, six down-regulated genes (phos-phoenolpyruvate carboxylase 4, sedoheptulose-bisphosphatase, ribose-5-phosphate isomerase 2, high cyclic electron flow 1, beta glucosidase 32 and starch synthase 2) and two up-regulated genes (beta glucosidase 2 and UDP-glucose pyrophosphorylase 2) implicated in photosynthetic carbon fixation and starch/sucrose metabolism were identified during the recovery process. To augment these insights, a weighted gene co-expression network analysis yielded a co-expression network, pinpointing the hub genes correlated with ChlF dynamics' variation trends. The cumulative results showed that HS inhibited the synthesis of photosynthesis-antenna proteins in R.×pulchrum leaves. This disruption subsequently led to diminished photochemical activities in both PSII and PSI, albeit with PSI exhibiting heightened thermostability. Depending on the regulation of the reactive oxygen species scavenging system and heat dissipation, photoprotection sustained the recoverability of R. × pulchrum to HS.

Copper and zinc accumulation, distribution, and tolerance in Pistia stratiotes L.; revealing the role of root caps

Pollution by potentially toxic trace metals, such as copper or zinc, is global. Both Cu and Zn are essential microelements, which in higher concentrations become toxic. The aquatic plant Pistia stratiotes(L. has great potential for phytoremediation. Also, it has an unusually large and easily detachable root cap, which makes it a suitable model for studying the potential role of the root cap in metal uptake. Plant response to environmentally relevant concentrations of Cu (0.1, 0.3, and 1 μM) and Zn (0.3, 1, and 3 μM) was investigated with the aim of studying their interaction and distribution at the root tissue level as well as revealing their tolerance mechanisms. Changes in the root anatomy and plant ionome were determined using light and fluorescence microscopy, ICP-MS, and μXRF imaging. Alterations in photosynthetic activity caused by Cu or Zn excesses were monitored by direct imaging of fast chlorophyll fluorescence kinetics (OJIP). Fe and Mn were preferentially localized in the root cap, while Ca, Cu, Ni, and Zn were mainly in the root tip regardless of the Cu/Zn treatment. Translocation of Cu and Zn to the leaves increased with higher doses, however the translocation factor was the lowest in the highest treatments. Measurements of photosynthetic parameters showed a higher susceptibility of electron transport flux from QA to QB under increasing Cu than Zn supply. This, along with our findings regarding the root anatomy and the differences in Ca accumulation and distribution, led to the conclusion that P. stratiotes is more effective for Zn remediation than Cu.

Photosynthesis, Yield and Quality in Wild Rocket (Diplotaxis tenuifolia L.) under Photoluminescent Greenhouse Covers

Manipulation of light spectral composition is a useful tool to drive morphological, physiological and metabolic responses in several crops, ultimately improving yield and quality. Novel materials for greenhouse covering are being developed in order to make a better use of the available sunlight: among these are the cover films or panels incorporating fluorescent additives which are able to convert UV solar radiation into visible light. In this research, we compared the physiological traits and the agronomical performance of wild rocket grown in pots in the winter–spring season, under four different greenhouse prototypes covered with poly-methyl-methacrylate (PMMA)-based panels. PMMA panels doped at 3% (Dop3) or 7% (Dop7) w/w with a blend of rare-earth elements (partially converting the solar UV radiation to red and blue wavelengths) were compared with an undoped (UD) and a whitewashed (WW) PMMA greenhouse. The rocket yield was higher in Dop3 (+30%), while it was unaffected in Dop7 and lower in WW (−39%), compared to the control (6.06 kg m−2). The leaf greenness decreased while both the ABTS and the hydrophilic antioxidant activities increased under the doped and the whitewashed greenhouses. The Dop3 treatment provided the best results in terms of yield and quality of greenhouse wild rocket in winter–spring cycle. However, the analysis of OJIP kinetics of chlorophyll fluorescence revealed that the main factor affecting the photosynthetic performance was the light intensity inside each greenhouse rather than the modulation of light spectrum, because of the different shading properties of the doping and whitewashing treatments. Although these results did not allow us to distinguish between the combined effects of shading and light spectrum modulation, the use of photoluminescent covers can be foreseen as a promising innovation in greenhouse horticulture.

Cadmium and Zn hyperaccumulation provide efficient constitutive defense against Turnip yellow mosaic virus infection in Noccaea caerulescens

To understand the role of Zn and Cd in anti-viral defence, Zn/Cd hyperaccumulator Noccaea caerulescens plants grown with deficient (0.3 µM), replete (10 µM) and excess (100 µM) Zn2+ and Cd (10 µM Zn2+ + 1 µM Cd2+) were infected with Turnip yellow mosaic virus (TYMV). Gas exchange and chlorophyll fluorescence kinetics analyses demonstrated direct TYMV effects on photosynthetic light reactions but N. caerulescens was more resistant against TYMV than the previously studied non-hyperaccumulator N. ochroleucum. Virus abundance and photosynthesis inhibition were the lowest in the high Zn and Cd treatments. RNAseq analysis of 10 µM Zn2+ plants revealed TYMV-induced upregulation of Ca transporters, chloroplastic ZTP29 and defence genes, but none of those that are known to be strongly involved in hyperaccumulation. Synchrotron µ-XRF tomography, however, showed that Zn hyperaccumulation remained strongest in vacuoles of epidermal storage cells regardless of infection. This was in contrast to N. ochroleucum, where apoplastic Zn drastically increased in response to TYMV. These results suggest that the antiviral response of N. caerulescens is less induced by the onset of this biotic stress, but it is rather a permanent resistant state of the plant. Real-time qPCR revealed upregulation of ferritin in Zn10 infected plants, suggesting Fe deprivation as a virus defence strategy under suboptimal Zn supply.

Recording of Low-Oxygen Stress Response Using Chlorophyll Fluorescence Kinetics in Apple Fruit

AbstractLong-term storage of apples (Malus x domestica, Borkh.) is increasingly taking place under Dynamic Controlled Atmosphere (DCA). The oxygen level is lowered to ≤ 1 kPa O2 and the apples are stored just above the Lower Oxygen Limit (LOL). Low oxygen stress during controlled atmosphere storage can lead to fermentation in apples if oxygen levels are too low. Chlorophyll fluorescence can be used to detect low-oxygen stress at an early stage during storage. The currently available non-imaging fluorescence systems often use the minimal fluorescence (Fo) parameter. In contrast, the use of chlorophyll fluorescence kinetics is insufficiently described. Therefore, this study aimed to gain more knowledge about the response of chlorophyll fluorescence kinetics to low oxygen stress in apples using a fluorescence imaging system. The results show that the kinetic fluorescence curves differ under aerobic and fermentation conditions. The fermentative conditions initiated a decrease in fluorescence intensity upon application of the saturation pulses during exposure to actinic light. This result was made at 18 °C and 2 °C ambient temperatures. Interestingly, the kinetic curve changed at 2 °C before fermentation products accumulated in the apples. Non-photochemical quenching (NPQ) decreased under fermentation conditions in the dark phase after relaxation. Upon entering the dark relaxation phase after Kautsky induction, ɸPSII began to increase. Under atmospheric oxygen conditions, ɸPSII reached values of 0.81 to 0.76, while under fermentation, ɸPSII values ranged from 0.57 to 0.44.

Polyamine, 1,3-diaminopropane, regulates defence responses on growth, gas exchange, PSII photochemistry and antioxidant system in wheat under arsenic toxicity

The metalloid arsenic (As) is extremely hazardous to all living organisms, including plants. Pollution with As is very detrimental to the photosynthetic machinery, cell division, energy generation, and redox status. In order to cope with stress, the use of growth regulators such as polyamines (PA), which strengthen the antioxidant system of plants, has become widespread in recent years. PAs can modulate the plant growth through basic mechanisms common to all living organisms, such as membrane stabilization, free radical scavenging, deoxyribonucleic acid (DNA), ribonucleic acid (RNA) and protein synthesis, enzyme activities and second messengers. However, the effect of 1,3- diaminopropane (Dap), which is a product of PA catabolism, is not clear enough in plants exposed to As toxicity. In the current study, the different concentrations of 1,3-diaminopropane (0.1, 0.5 and 1 mM Dap) were hydroponically treated to wheat (Triticum aestivum) under arsenic stress (100 μM As) and then relative growth rate (RGR), relative water content (RWC), proline content (Pro), gas exchange parameters, PSII photochemistry, chlorophyll fluorescence kinetics, antioxidant activity and lipid peroxidation were assessed. RGR, RWC, osmotic potential and Pro content decreased in As-applied plants. The inhibition of these parameters could be reversed by Dap treatments. Besides, Dap applications mitigated the As toxicity-induced suppression on chlorophyll fluorescence (Fv/Fm, Fv/Fo and Fo/Fm) and the performance of PSII photochemistry. As impaired the balance on antioxidant capacity by decreased activities of catalase (CAT), peroxidase (POX), glutathione peroxidase (GPX), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), and the contents of ascorbate (AsA) and glutathione (GSH) and then lipid peroxidation (TBARS content) increased. In the presence of Dap under As stress, the plants exhibited an increase in superoxide dismutase (SOD), POX, and GPX. Dap treatments contributed to the maintenance of cellular redox state (AsA/DHA and GSH/GSSG) by regulating the activities/contents of enzyme/non-enzyme involved in the AsA-GSH cycle. After Dap applications against stress, ROS accumulation (H2O2 content) and lipid peroxidation (TBARS) were effectively reduced. The findings showed that by eliminating As-induced oxidative damage and protecting the biochemical processes of photosynthesis, Dap treatments have a substantial potential to give resistance to wheat.

Derivative-based time-adjusted analysis of diurnal and within-tree variation in the OJIP fluorescence transient of silver birch

The JIP test, based on fast chlorophyll fluorescence (ChlF) kinetics and derived parameters, is a dependable tool for studying photosynthetic efficiency under varying environmental conditions. We extracted additional information from the whole OJIP and the normalized variable fluorescence (Vt) transient curve using first and second-order derivatives to visualize and localize points of landmark events. To account for light-induced variations in the fluorescence transient, we present a time-adjusted JIP test approach in which the derivatives of the transient curve are used to determine the exact timing of the J and I steps instead of fixed time points. We compared the traditional JIP test method with the time-adjusted method in analyzing fast ChlF measurements of silver birch (Betula pendula) in field conditions studying diurnal and within-crown variation. The time-adjusted JIP test method showed potential for studying ChlF dynamics, as it takes into account potential time shifts in the occurrence of J and I steps. The exact occurrence times of J and I steps and other landmark events coincided with the times of significant differences in fluorescence intensity. Chlorophyll fluorescence parameters were linearly related to photosynthetic photon flux density (PPFD) at different times of day, and the values obtained by the time-adjusted JIP test showed a stronger linear regression than the traditional JIP test. For fluorescence parameters having significant differences among different times of day and crown layers, the time-adjusted JIP test resulted in more clear differences than the traditional JIP test. Diurnal ChlF intensity data indicated that differences between the southern and northern provenance were only evident under low light conditions. Taken together, our results emphasize the potential relevance of considering the time domain in the analysis of the fast ChlF induction.

The Effects of Growth Modification on Pollen Development in Spring Barley (Hordeum vulgare L.) Genotypes with Contrasting Drought Tolerance

Drought stress inducing pollen sterility can reduce crop yield worldwide. The regulatory crosstalk associated with the effects of drought on pollen formation at the cellular level has not been explored in detail so far. In this study, we performed morphological and cytoembryological analysis of anther perturbations and examined pollen development in two spring barley genotypes that differ in earliness and drought tolerance. The Syrian breeding line CamB (drought-tolerant) and the European cultivar Lubuski (drought-sensitive) were used as experimental materials to analyze the drought-induced changes in yield performance, chlorophyll fluorescence kinetics, the pollen grain micromorphology and ultrastructure during critical stages of plant development. In addition, fluctuations in HvGAMYB expression were studied, as this transcription factor is closely associated with the development of the anther. In the experiments, the studied plants were affected by drought, as was confirmed by the analyses of yield performance and chlorophyll fluorescence kinetics. However, contrary to our expectations, the pollen development of plants grown under specific conditions was not severely affected. The results also suggest that growth modification, as well as the perturbation in light distribution, can affect the HvGAMYB expression. This study demonstrated that the duration of the vegetation period can influence plant drought responses and, as a consequence, the processes associated with pollen development as every growth modification changes the dynamics of drought effects as well as the duration of plant exposition to drought.

Halotolerant plant growth-promoting bacteria, Bacillus pumilus, modulates water status, chlorophyll fluorescence kinetics and antioxidant balance in salt and/or arsenic-exposed wheat

Seed priming is an effective and novel technique and the use of eco-friendly biological agents improves the physiological functioning in the vegetative stage of plants. This procedure ensures productivity and acquired stress resilience in plants against adverse conditions without contaminating the environment. Though the mechanisms of bio-priming-triggered alterations have been widely explained under induvial stress conditions, the interaction of combined stress conditions on the defense system and the functionality of photosynthetic apparatus in the vegetative stage after the inoculation to seeds has not been fully elucidated. After Bacillus pumilus inoculation to wheat seeds (Triticum aestivum), three-week-old plants were hydroponically exposed to the alone and combination of salt (100 mM NaCl) and 200 μM sodium arsenate (Na2HAsO4·7H2O, As) for 72 h. Salinity and As pollutant resulted in a decline in growth, water content, gas exchange parameters, fluorescence kinetics and performance of photosystem II (PSII). On the other hand, the seed inoculation against stress provided the alleviation of relative growth rate (RGR), relative water content (RWC) and chlorophyll fluorescence. Since there was no effective antioxidant capacity, As and/or salinity caused the induction of H2O2 accumulation and thiobarbituric acid reactive substances content (TBARS) in wheat . The inoculated seedlings had a high activity of superoxide dismutase (SOD) under stress. B. pumilis decreased the NaCl-induced toxic H2O2 levels by increasing peroxidase (POX) and enzymes/non-enzymes related to ascorbate-glutathione (AsA-GSH) cycle. In the presence of As exposure, the inoculated plants exhibited an induction in CAT activity. On the other hand, for H2O2 scavenging, the improvement in the AsA-GSH cycle was observed in bacterium priming plants plus the combined stress treatment. Since B. pumilus inoculation reduced H2O2 levels against all stress treatments, lipid peroxidation subsequently decreased in wheat leaves. The findings obtained from our study explained that the seed inoculation with B. pumilus provided an activation in the defense system and protection in growth, water status, and gas exchange regulation in wheat plants against the combination of salt and As.

The effects of heading time on yield performance and HvGAMYB expression in spring barley subjected to drought

In the lifetime of a plant, flowering is not only an essential part of the reproductive process but also a critical developmental stage that can be vulnerable to environmental stresses. To ensure survival during drought, plants accelerate the flowering process, and this response is known as “drought escape.” HvGAMYB—transcription factor associated, among others, with flowering process and anther development in barley—has also an important role in developmental modification and yield performance in plants subjected to stressed conditions. Due to the fact that information about the mechanisms associated both with the flowering acceleration and the anther or pollen disruption is limited, the exploration of the potential HvGAMYB role in flower development may shed light on pollen and spike morphology formations in plants grown under unfavorable water conditions. The aim of this study was to characterize differences in responses to drought among early- and late-heading barley genotypes. These two subgroups of plants—differentiated in terms of phenology—were analyzed, and traits linked to plant phenotype, physiology, and yield were investigated. In our study, the drought stress reactions of two barley subgroups showed a wide range of diversity in terms of yield performance, anther morphology, chlorophyll fluorescence kinetics, and pollen viability. The studied plants exhibited different yield performances under control and drought conditions. Moreover, the random distribution of genotypes on the biplot showing variability of OJIP parameters in the second developmental point of our investigation revealed that prolonged drought stress caused that among early- and late-heading plants, the studied genotypes exhibited different responses to applied stress conditions. The results of this study also showed that the HvGAMYB expression level was correlated positively with traits associated with lateral spike morphology in the second developmental point of this investigation, which showed that this association occurred only under prolonged drought and highlighted the drought stress duration effect on the HvGAMYB expression level.

Impacts of Radio-Frequency Electromagnetic Field (RF-EMF) on Lettuce (Lactuca sativa)—Evidence for RF-EMF Interference with Plant Stress Responses

The increased use of wireless technology causes a significant exposure increase for all living organisms to radio frequency electromagnetic fields (RF-EMF). This comprises bacteria, animals, and also plants. Unfortunately, our understanding of how RF-EMF influences plants and plant physiology remains inadequate. In this study, we examined the effects of RF-EMF radiation on lettuce plants (Lactuca sativa) in both indoor and outdoor environments using the frequency ranges of 1890-1900 MHz (DECT) at 2.4 GHz and 5 GHz (Wi-Fi). Under greenhouse conditions, RF-EMF exposure had only a minor impact on fast chlorophyll fluorescence kinetics and no effect on plant flowering time. In contrast, lettuce plants exposed to RF-EMF in the field showed a significant and systemic decrease in photosynthetic efficiency and accelerated flowering time compared to the control groups. Gene expression analysis revealed significant down-regulation of two stress-related genes in RF-EMF-exposed plants: violaxanthin de-epoxidase (VDE) and zeaxanthin epoxidase (ZEP). RF-EMF-exposed plants had lower Photosystem II's maximal photochemical quantum yield (FV/FM) and non-photochemical quenching (NPQ) than control plants under light stress conditions. In summary, our results imply that RF-EMF might interfere with plant stress responses and reduced plant stress tolerance.

Responses of individual and combined polystyrene and polymethyl methacrylate nanoplastics on hormonal content, fluorescence/photochemistry of chlorophylls and ROS scavenging capacity in Lemna minor under arsenic-induced oxidative stress

Nanoplastics alter the adverse impacts of hazardous contaminants such as heavy metals by changing their adsorption and accumulation. Few findings are available on the interaction between nanoplastic and heavy metals in plants. However, there is no report on the mechanisms for removing metal stress-mediated oxidative damage by the combination treatments of nanoplastics. To address this lack of information, polystyrene nanoplastic (PS, 100 mg L-1) and polymethyl methacrylate (PMMA, 100 mg L-1) were hydroponically applied to Lemna minor exposed to arsenate (As, 100 μM) for 7 days. PS or PMMA caused a reduction in the contents of N, P, K, Ca, Mg and Mn, but the improved contents were detected in the presence of PS or PMMA plus As stress. The hormone contents (auxin, gibberellic acid, cytokinin, salicylic acid and jasmonic acid) reduced by stress were re-arranged through PS or PMMA applications. Based on chlorophyll efficiency, fluorescence kinetics and performance of PSII, the impaired photosynthesis by As stress was improved via PS or PMMA applications. This alleviation did not continue under the combined form of PS and PMMA in As-applied plants. All analyzed antioxidant activity (superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX), glutathione reductase (GR), glutathione S-transferase (GST), glutathione peroxidase (GPX), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR)) decreased or unchanged under As, PS or PMMA. Due to the inactivation of the defense system, L. minor had high levels of hydrogen peroxide (H2O2) and thiobarbituric acid reactive substances (TBARS), showing lipid peroxidation. After As toxicity, induvial applications of PS or PMMA indicated the activated enzyme capacity (SOD, POX, GST and GPX) and upregulated AsA/DHA, GSH/GSSG and redox state of GSH, which facilitated the removal of radical accumulation. The efficiency of the antioxidant system in As + PS + PMMA-applied L. minor was not enough to remove damage induced by As stress; hereby, TBARS and H2O2 contents were similar to the As-treated group. Our findings from alone or combined application of PS and PMMA provide new information to advance the tolerance mechanism against As exposure in L. minor.

Acclimation Response of Green Microalgae Chlorella Sorokiniana to 2,3',4,4',6-Pentachlorobiphenyl.

The effect of the toxicant 2,3',4,4',6-pentachlorobiphenyl (PCB-119) on the growth, chlorophyll content, and PSII activity of C. sorokiniana cells was investigated. A strong negative effect of the toxicant was observed at PCB concentration of 0.05 μg mL-1 , when culture growth ceased, chlorophyll strongly bleached, and cell death occurred. The use of original highly sensitive fluorimeter to measure three types of high-resolution chlorophyll fluorescence kinetics allowed us to detect an initial dramatic decrease in the activity of primary photosynthetic reactions, followed by their almost complete recovery at the end of the incubation period when most cells were dead. The study of the distribution of individual cells in culture in terms of Fv /Fm parameter, which reflects the quantum yield of PSII photochemistry, revealed the existence of 2-3% of cells retaining high Fv /Fm (>0.7) in the presence of the toxicant. The treated cultures were able to resume growth after prolonged incubation in fresh medium. The high sensitivity fluorescence methods used made it possible to identify stress-resistant cells which maintain high photosynthetic activity in the presence of lethal doses of toxic substances; these cells provide recovery of the population after stress.

Chlorophyll fluorometry in evaluating photosynthetic performance: key limitations, possibilities, perspectives and alternatives.

Non-destructive methods for theassessment of photosynthetic parameters of plants are widely applied to evaluate rapidly the photosynthetic performance, plant health, and shifts in plant productivity induced by environmental and cultivation conditions. Most of these methods are based on measurements of chlorophyll fluorescence kinetics, particularly on pulse modulation (PAM) fluorometry. In this paper, fluorescence methods are critically discussed in regard to some their possibilities and limitations inherent to vascular plants and microalgae. Attention is paid to the potential errors related to the underestimation of thylakoidal cyclic electron transport and anoxygenic photosynthesis. PAM-methods are also observed considering the color-addressed measurements. Photoacoustic methods are discussed as an alternative and supplement to fluorometry. Novel Fourier modifications of PAM-fluorometry and photoacoustics are noted as tools allowing simultaneous application of a dual or multi frequency measuring light for one sample.