Search Results (31)

Background: Gallbladder hypomotility is a key pathogenic factor in cholelithiasis. Non-invasive interventions to enhance gallbladder contractility remain limited. Ultrasound therapy has shown promise in various muscular disorders, but its effects on gallbladder function are unexplored. Methods: This study employed low-intensity pulsed ultrasound (LIPUS) at a 3 MHz frequency and 0.8 W/cm² intensity with a 20% duty cycle to irradiate the gallbladder region of fasting guinea pigs. Gallbladder contractile function was evaluated through multiple complementary approaches: in vivo assessment via two-dimensional/three-dimensional ultrasound imaging to monitor volumetric changes; quantitative functional evaluation using nuclear medicine scintigraphy (^99mTc-HIDA); and ex vivo experiments including isolated gallbladder muscle strip tension measurements, histopathological analysis, α-smooth muscle actin (α-SMA) immunohistochemistry, and intracellular calcium fluorescence imaging. Results: Ultrasound significantly enhanced gallbladder emptying, evidenced by the volume reduction and increased ejection fraction. Scintigraphy confirmed accelerated bile transport in treated animals. Ex vivo analyses demonstrated augmented contractile force, amplitude, and frequency in ultrasound-treated smooth muscle. Histological examination revealed smooth muscle hypertrophy, α-SMA upregulation, and elevated intracellular calcium levels. Extended ultrasound exposure produced sustained functional improvements without tissue damage. Conclusions: Ultrasound effectively enhances gallbladder contractile function through mechanisms involving smooth muscle structural modification and calcium signaling modulation. These findings establish the experimental foundation for ultrasound as a promising non-invasive therapeutic approach to improve gallbladder motility and potentially prevent gallstone formation. Full article

The fraction of open Photosystem II (PSII) reaction centers (q_L) is critical for connecting broadband PSII fluorescence (ChlF_PSII) with the actual electron transport from PSII to Photosystem I. Accurately estimating q_L is fundamental for determining ChlF_PSII, which, in turn, is vital for mechanistically estimating the actual electron transport rate and photosynthetic CO₂ assimilation. Chlorophyll fluorescence provides direct physiological insights, offering a robust foundation for q_L estimation. However, uncertainties in the ChlF_PSII–q_L relationship across different plant functional types (PFTs) limit its broader application at large spatial scales. To address this issue, we developed a leaf-level instrument capable of simultaneously measuring actively and passively induced chlorophyll fluorescence. Using this system, we measured light response, CO₂ response, and temperature response curves across 52 species representing seven PFTs. Our findings reveal the following: (1) a strong linear correlation between ChlF_PSII derived from passively induced fluorescence and that from actively induced fluorescence (R² = 0.85), and (2) while the parameters of the ChlF_PSII–q_L relationship varied among PFTs, ChlF_PSII reliably modeled q_L within each PFT, with the R² ranging from 0.85 to 0.96. This study establishes quantitative ChlF_PSII–q_L relationships for various PFTs by utilizing passively induced fluorescence to calculate ChlF_PSII. The results demonstrate the potential for remotely sensed chlorophyll fluorescence data to estimate q_L and strengthen the use of fluorescence-based approaches for mechanistic GPP estimation at large spatial scales. Full article

Sargassum is an important primary producer of rocky bottom communities in coastal ecosystems. Like other parts of the planet, benthic populations of S. natans from Ilha Grande Bay (IGB), southeastern Brazil, have been suffering from different forms of natural and anthropogenic disturbances, in particular increasing seawater temperatures. The aim of this study was to understand the effects of temperature on the photosynthetic performance of S. natans using the pulse amplitude modulated (PAM) fluorometry. In the field experiments, the occurrence of photoprotection resulted in a difference between the effective and maximum quantum yields [(ΔF (F’_m − F_s)/F’_m and F_v/F_m, respectively) that was maximized at noon. The stress induced by incubation at 32–35 °C caused a decrease in F_v/F_m by 33% on the first day and approximately 20% on subsequent days. In the laboratory, using two co-occurred species of S. natans and Padina gymnospora, we verified that the photosynthetic apparatus of S. natans collapses at 34 °C. The fate of the energy absorbed by photosystem II (PSII) antenna showed that, in S. natans, photochemical activity and non-photochemical quenching of chlorophyll fluorescence (NPQ) drastically decrease, and only the passive dissipation in the form of heat and fluorescence remains. Our results indicate the disappearance of the NPQ photoprotection at 34 °C before the decline of F_v/F_m as the reason for the collapse of photochemistry of Sargassum. Full article

In addition to leaves, photosynthesis can occur in other green plant organs, including developing seeds of many crops. While the majority of studies examining photosynthesis are concentrated on the leaf level, the role of other green tissues in the production of total photoassimilates has been largely overlooked. The present work studies the photosynthetic behavior of leaves and non-foliar (pericarps, coats, and cotyledons) organs of pea (Pisum sativum L.) plants at the middle stage of seed maturation. The Chl a fluorescence transient was examined based on OJIP kinetics using the FluorPen FP 110. A discrepancy was observed between the performance index (PI_ABS) for foliar and non-foliar plant tissues, with the highest level noted in the leaves. The number of absorbed photons (ABS) and captured energy flow (TRo) per reaction center (RC) were elevated in the non-foliar tissues, which resulted in a faster reduction in Q_A. Conversely, the energy dissipation flux per RC (DIo/RC and PHI_Do) indicated an increase in the overall dissipation potential of active reaction centers of photosystem II. This phenomenon was attributed to the presence of a higher number of inactive RCs in tissues that had developed under low light intensity. Furthermore, the expression of genes associated with proteins and enzymes that regulate ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCo) activity was observed, including chaperonins Cpn60α and Cpn60β, RuBisCO activase, as well as phosphoribulokinase. The expression of these genes was found to differ between foliar and non-foliar tissues, indicating that the activation state of RuBisCO may be modified in response to light intensity. Overall, the present study provides insights into the mechanisms by which non-foliar green tissues of plants adapt to efficient light capture and utilization under low light conditions. Full article

The constantly growing need to increase the production of agricultural products in changing climatic conditions makes it necessary to accelerate the development of new cultivars that meet the modern demands of agronomists. Currently, the breeding process includes the stages of genotyping and phenotyping to optimize the selection of promising genotypes. One of the most popular phenotypic methods is the pulse-amplitude modulated (PAM) fluorometry, due to its non-invasiveness and high information content. In this review, we focused on the opportunities of using chlorophyll fluorescence (ChlF) parameters recorded using PAM fluorometry to assess the state of plants in drought and heat stress conditions and predict the economically significant traits of wheat, as one of the most important agricultural crops, and also analyzed the relationship between the ChlF parameters and genetic markers. Full article

Winter oilseed rape (Brassica napus L.), Europe’s foremost oilseed crop, is significantly impacted by hailstorms, leading to substantial yield reductions that are difficult to predict and measure using conventional methods. This research aimed to assess the effectiveness of photosynthetic efficiency analysis for predicting yield loss in winter rapeseed subjected to hail exposure. The aim was to pinpoint the chlorophyll fluorescence parameters most affected by hail stress and identify those that could act as non-invasive biomarkers of yield loss. The study was conducted in partially controlled conditions (greenhouse). Stress was induced in the plants by firing plastic balls with a 6 mm diameter at them using a pneumatic device, which launched the projectiles at speeds of several tens of meters per second. Measurements of both continuous-excitation and pulse-modulated-amplitude chlorophyll fluorescence were engaged to highlight the sensitivity of the induction curve and related parameters to hail stress. Our research uncovered that some parameters such as F_s, F_m’, Φ_PSII, ETR, F_o, F_v/F_m, and F_v/F_o measured eight days after the application of stress had a strong correlation with final yield, thus laying the groundwork for the creation of new practical protocols in agriculture and the insurance industry to accurately forecast damage to rapeseed crops due to hail stress. Full article

Early detection of pathogens can significantly reduce yield losses and improve the quality of agricultural products. This study compares the efficiency of hyperspectral (HS) imaging and pulse amplitude modulation (PAM) fluorometry to detect pathogens in plants. Reflectance spectra, normalized indices, and fluorescence parameters were studied in healthy and infected areas of leaves. Potato virus X with GFP fluorescent protein was used to assess the spread of infection throughout the plant. The study found that infection increased the reflectance of leaves in certain wavelength ranges. Analysis of the normalized reflectance indices (NRIs) revealed indices that were sensitive and insensitive to infection. NRI_700/850 was optimal for virus detection; significant differences were detected on the 4th day after the virus arrived in the leaf. Maximum (F_v/F_m) and effective quantum yields of photosystem II (Φ_PSII) and non-photochemical fluorescence quenching (NPQ) were almost unchanged at the early stage of infection. Φ_PSII and NPQ in the transition state (a short time after actinic light was switched on) showed high sensitivity to infection. The higher sensitivity of PAM compared to HS imaging may be due to the possibility of assessing the physiological changes earlier than changes in leaf structure. Full article

Solar-induced chlorophyll fluorescence (SIF) has a high correlation with Gross Primary Production (GPP). However, studies focusing on the impact of drought on the SIF-GPP relationship have had mixed results at various scales, and the mechanisms controlling the dynamics between photosynthesis and fluorescence emission under water stress are not well understood. We developed a leaf-scale measurement system to perform concurrent measurements of active and passive fluorescence, and gas-exchange rates for winter wheat experiencing a one-month progressive drought. Our results confirmed that: (1) shifts in light energy allocation towards decreasing photochemistry (the quantum yields of photochemical quenching in PSII decreased from 0.42 to 0.21 under intermediate light conditions) and increasing fluorescence emissions (the quantum yields of fluorescence increased to 0.062 from 0.024) as drought progressed enhance the degree of nonlinearity of the SIF-GPP relationship, and (2) SIF alone has a limited capacity to track changes in the photosynthetic status of plants under drought conditions. However, by incorporating the water stress factor into a SIF-based mechanistic photosynthesis model, we show that drought-induced variations in a variety of key photosynthetic parameters, including stomatal conductance and photosynthetic CO₂ assimilation, can be accurately estimated using measurements of SIF, photosynthetically active radiation, air temperature, and soil moisture as inputs. Our findings provide the experimental and theoretical foundations necessary for employing SIF mechanistically to estimate plant photosynthetic activity during periods of drought stress. Full article

Non-photochemical quenching (NPQ) is an indicator of crop stress. Until now, only a limited number of studies have focused on how to estimate NPQ using remote sensing technology. The main challenge is the complicated regulatory mechanism of NPQ. NPQ can be divided into energy-dependent (qE) and non-energy-dependent (non-qE) quenching. The contribution of these two components varies with environmental factors, such as light intensity and stress level due to the different response mechanisms. This study aims to explore the feasibility of estimating NPQ using photosynthesis-related vegetation parameters available from remote sensing by considering the two components of NPQ. We concurrently measured passive vegetation reflectance spectra by spectrometer, as well as active fluorescence parameters by pulse-amplitude modulated (PAM) of rice (Oryza sativa) leaves. Subsequently, we explored the ability of the selected vegetation parameters (including the photochemical reflectance index (PRI), inverted red-edge chlorophyll index (IRECI), near-infrared reflectance of vegetation (NIR_v), and fluorescence quantum yield (ΦF)) to estimate NPQ. Based on different combinations of these remote sensing parameters, empirical models were established to estimate NPQ using the linear regression method. Experimental analysis shows that the contribution of qE and non-qE components varied under different illumination conditions. Under high illumination, the NPQ was attributed primarily to the qE component, while under low illumination, it was equally attributed to the qE and non-qE components. Among all tested parameters, ΦF was sensitive to the qE component variation, while IRECI and NIR_v were sensitive to the non-qE component variation. Under high illumination, integrating ΦF in the regression model captured NPQ variations well (R² > 0.74). Under low illumination, ΦF, IRECI, and NIR_v explained 24%, 62%, and 65% of the variation in NPQ, respectively, while coupling IRECI or NIR_v with ΦF considerably improved the accuracy of NPQ estimation (R² > 0.9). For all the samples under both low and high illumination, the combination of ΦF with at least one of the other parameters (including IRECI, NIR_v and PAR) offers a more versatile and reliable approach to estimating NPQ than using any single parameter alone. The findings of this study contribute to the further development of remote sensing methods for NPQ estimation at the canopy scale in the future. Full article

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. Full article

Cyanine fluorophores are extensively used in fluorescence spectroscopy and imaging. Upon continuous excitation, especially at excitation conditions used in single-molecule and super-resolution experiments, photo-isomerized states of cyanines easily reach population probabilities of around 50%. Still, effects of photo-isomerization are largely ignored in such experiments. Here, we studied the photo-isomerization of the pentamethine cyanine 5 (Cy5) by two similar, yet complementary means to follow fluorophore blinking dynamics: fluorescence correlation spectroscopy (FCS) and transient-state (TRAST) excitation–modulation spectroscopy. Additionally, we combined TRAST and spectrofluorimetry (spectral-TRAST), whereby the emission spectra of Cy5 were recorded upon different rectangular pulse-train excitations. We also developed a framework for analyzing transitions between multiple emissive states in FCS and TRAST experiments, how the brightness of the different states is weighted, and what initial conditions that apply. Our FCS, TRAST, and spectral-TRAST experiments showed significant differences in dark-state relaxation amplitudes for different spectral detection ranges, which we attribute to an additional red-shifted, emissive photo-isomerized state of Cy5, not previously considered in FCS and single-molecule experiments. The photo-isomerization kinetics of this state indicate that it is formed under moderate excitation conditions, and its population and emission may thus deserve also more general consideration in fluorescence imaging and spectroscopy experiments. Full article

Drought, the major limiting factor for plant growth and crop productivity, affecting several physiological and biochemical processes, is expected to increase in duration, intensity, and frequency as a consequence of climate change. Plants have developed several approaches to either avoid or tolerate water deficit. Plants as a response to drought stress (DS), close stomata, reducing carbon dioxide (CO₂) entry in the leaf, thus decreasing photosynthesis which results in reduced synthesis of essential organic molecules that sustain the life on earth. The reduced CO₂ fixation, decreases electron transport rate (ETR), while the absorbed light energy overdoes what can be used for photochemistry resulting in excess reactive oxygen species (ROS) and oxidative stress. Current imaging techniques allow non-destructive monitoring of changes in the physiological state of plants under DS. Thermographic visualization, near-infrared imaging, and chlorophyll a fluorescence imaging are the most common verified imaging techniques for detecting stress-related changes in the display of light emission from plant leaves. Chlorophyll a fluorescence analysis, by use of the pulse amplitude modulation (PAM) method, can principally calculate the amount of absorbed light energy that is directed for photochemistry in photosystem II (PSII) (Φ_PSII), dissipated as heat (Φ_NPQ), or dissipated by the non-radiative fluorescence processes (Φ_NO). The method of chlorophyll a fluorescence imaging analysis by providing colour pictures of the whole leaf PSII photochemistry, can successfully identify the early drought stress warning signals. Its implementation allowed visualization of the leaf spatial photosynthetic heterogeneity and discrimination between mild drought stress (MiDS), moderate drought stress (MoDS), and severe drought stress (SDS). The fraction of open reaction centers of PSII (qp) is suggested as the most sensitive and suitable indicator of an early drought stress warning and also for selecting drought tolerant cultivars. Full article

Dormancy is a physiological state that confers winter hardiness to and orchestrates phenological phase progression in temperate perennial plants. Weather fluctuations caused by climate change increasingly disturb dormancy onset and release in plants including tree crops, causing aberrant growth, flowering and fruiting. Research in this field suffers from the lack of affordable non-invasive methods for online dormancy monitoring. We propose an automatic framework for low-cost, long-term, scalable dormancy studies in deciduous plants. It is based on continuous sensing of the photosynthetic activity of shoots via pulse-amplitude-modulated chlorophyll fluorescence sensors connected remotely to a data processing system. The resulting high-resolution time series of JIP-test parameters indicative of the responsiveness of the photosynthetic apparatus to environmental stimuli were subjected to frequency-domain analysis. The proposed approach overcomes the variance coming from diurnal changes of insolation and provides hints on the depth of dormancy. Our approach was validated over three seasons in an apple (Malus × domestica Borkh.) orchard by collating the non-invasive estimations with the results of traditional methods (growing of the cuttings obtained from the trees at different phases of dormancy) and the output of chilling requirement models. We discuss the advantages of the proposed monitoring framework such as prompt detection of frost damage along with its potential limitations. Full article

In the diatom Phaeodactylum tricornutum, iron limitation promotes a decrease in the content of photosystem II, as determined by measurements of oxygen-evolving activity, thermoluminescence, chlorophyll fluorescence analyses and protein quantification methods. Thermoluminescence experiments also indicate that iron limitation induces subtle changes in the energetics of the recombination reaction between reduced Q_B and the S₂/S₃ states of the water-splitting machinery. However, electron transfer from Q_A to Q_B, involving non-heme iron, seems not to be significantly inhibited. Moreover, iron deficiency promotes a severe decrease in the content of the extrinsic PsbV/cytochrome c₅₅₀ subunit of photosystem II, which appears in eukaryotic algae from the red photosynthetic lineage (including diatoms) but is absent in green algae and plants. The decline in the content of cytochrome c₅₅₀ under iron-limiting conditions is accompanied by a decrease in the binding of this protein to photosystem II, and also of the extrinsic PsbO subunit. We propose that the lack of cytochrome c₅₅₀, induced by iron deficiency, specifically affects the binding of other extrinsic subunits of photosystem II, as previously described in cyanobacterial PsbV mutants. Full article

Natural photonic structures are common across the biological kingdoms, serving a diversity of functionalities. The study of implications of photonic structures in plants and other phototrophic organisms is still hampered by missing methodologies for determining in situ photonic properties, particularly in the context of constantly adapting photosynthetic systems controlled by acclimation mechanisms on the cellular scale. We describe an innovative approach to determining spatial and spectral photonic properties and photosynthesis activity, employing micro-Fourier Image Spectroscopy and Pulse Amplitude Modulated Chlorophyll Fluorimetry in a combined microscope setup. Using two examples from the photosynthetic realm, the dynamic Bragg-stack-like thylakoid structures of Begonia sp. and complex 2.5 D photonic crystal slabs from the diatom Coscinodiscus granii, we demonstrate how the setup can be used for measuring self-adapting photonic-photosynthetic systems and photonic properties on single-cell scales. We suggest that the setup is well-suited for the determination of photonic–photosynthetic systems in a diversity of organisms, facilitating the cellular, temporal, spectral and angular resolution of both light distribution and combined chlorophyll fluorescence determination. As the catalogue of photonic structure from photosynthetic organisms is rich and diverse in examples, a deepened study could inspire the design of novel optical- and light-harvesting technologies. Full article