Search Results (18)

Chlorella is a rich source of nutrients. In addition to its nutritional value, it exhibits versatile biological activities. New strains have been extensively identified and investigated in recent years to expand the potential of Chlorella. The accurate measurement of pulse-amplitude-modulated (PAM) fluorometry parameters and effective microalgal cell lysis are foundational for advanced studies of novel Chlorella species. In this study, ribosomal small subunit (SSU)-internal transcribed spacer (ITS) phylogenetic analysis and internal transcribed spacer 2 (ITS2) secondary structure analysis were employed to identify a new Chlorella species. The dark adaptation time, the duration of the saturation pulse, the intensity of actinic light, and the duration of actinic light exposure for PAM fluorometry measurements were optimized. Different conditions of ultrasonication and high-pressure homogenization (HPH) for microalgal cell lysis were compared. Chlorella vulgaris K-01 was identified. The suitable duration for dark adaptation, the saturation pulse, and the actinic light were 15 min, 200 milliseconds, and 30 s, respectively. The suitable intensity of actinic light was 191 μE/(m²·s). For microalgal cell lysis, HPH could achieve 98.65% cell lysis efficiency at 30 kpsi (207 MPa), whereas ultrasonication attained an efficiency of 45.47% (300 W for 30 min). These results facilitate further study on the physiology and the composition of Chlorella vulgaris K-01. 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

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

The use of glyphosate-based herbicides (GBHs) worldwide has increased exponentially over the last two decades increasing the environmental risk to marine and coastal habitats. The present study investigated the effects of GBHs at environmentally relevant concentrations (0, 10, 50, 100, 250, and 500 μg·L⁻¹) on the physiology and biochemistry (photosynthesis, pigment, and lipid composition, antioxidative systems and energy balance) of Ulva lactuca, a cosmopolitan marine macroalgae species. Although GBHs cause deleterious effects such as the inhibition of photosynthetic activity, particularly at 250 μg·L⁻¹, due to the impairment of the electron transport in the chloroplasts, these changes are almost completely reverted at the highest concentration (500 μg·L⁻¹). This could be related to the induction of tolerance mechanisms at a certain threshold or tipping point. While no changes occurred in the energy balance, an increase in the pigment antheraxanthin is observed jointly with an increase in ascorbate peroxidase activity. These mechanisms might have contributed to protecting thylakoids against excess radiation and the increase in reactive oxygen species, associated with stress conditions, as no increase in lipid peroxidation products was observed. Furthermore, changes in the fatty acids profile, usually attributed to the induction of plant stress response mechanisms, demonstrated the high resilience of this macroalgae. Notably, the application of bio-optical tools in ecotoxicology, such as pulse amplitude modulated (PAM) fluorometry and laser-induced fluorescence (LIF), allowed separation of the control samples and those treated by GBHs in different concentrations with a high degree of accuracy, with PAM more accurate in identifying the different treatments. Full article

Coral reefs have been detrimentally impacted causing health issues due to elevated ocean temperatures as a result of increased greenhouse gases. Extreme temperatures have also exacerbated coral diseases in tropical reef environments. Numerous studies have outlined the impacts of thermal stress and disease on coral organisms, as well as understanding the influence of site-based characteristics on coral physiology. However, few have discussed the interaction of all three. Laboratory out-planting restoration projects have been of importance throughout impacted areas such as the Caribbean and southern Florida in order to increase coral cover in these areas. This study analyzes photosynthetic efficiency of Porites astreoides from the lower Florida Keys after a two-year reciprocal transplant study at inshore (Birthday reef) and offshore (Acer24 reef) sites to understand acclimation capacity of this species. Laboratory experiments subjected these colonies to one of three treatments: control conditions, increases in temperature, and increases in temperature plus exposure to an immune stimulant (lipopolysaccharide (LPS)) to determine their influence on photosynthetic efficiency and how stress events impact these measurements. In addition, this study is a continuation of previous studies from this group. Here, we aim to understand if these results are static or if an acclimation capacity could be found. Overall, we observed site-specific influences from the Acer24 reef site, which had significant decreases in photosynthetic efficiencies in 32 °C treatments compared to Birthday reef colonies. We suggest that high irradiance and lack of an annual recovery period from the Acer24 site exposes these colonies to significant photoinhibition. In addition, we observed significant increases in photosynthetic efficiencies from LPS exposure. We suggest host-derived antioxidants can mitigate the negative impacts of increased thermal stress. Further research is required to understand the full complexity of host immunity and symbiont photosynthetic interactions. Full article

Climate change-related events, such as marine heatwaves, are increasing seawater temperatures, thereby putting pressure on marine biota. The cosmopolitan distribution and significant contribution to marine primary production by the genus Ruppia makes them interesting organisms to study thermal tolerance and local adaptation. In this study, we investigated the photosynthetic responses in Ruppia to the predicted future warming in two contrasting bioregions, temperate Sweden and tropical Thailand. Through DNA barcoding, specimens were determined to Ruppia cirrhosa for Sweden and Ruppia maritima for Thailand. Photosynthetic responses were assessed using pulse amplitude-modulated fluorometry, firstly in short time incubations at 18, 23, 28, and 33 °C in the Swedish set-up and 28, 33, 38, and 43 °C in the Thai set-up. Subsequent experiments were conducted to compare the short time effects to longer, five-day incubations in 28 °C for Swedish plants and 40 °C for Thai plants. Swedish R. cirrhosa displayed minor response, while Thai R. maritima was more sensitive to both direct and prolonged temperature stress with a drastic decrease in the photosynthetic parameters leading to mortality. The results indicate that in predicted warming scenarios, Swedish R. cirrhosa may sustain an efficient photosynthesis and potentially outcompete more heat-sensitive species. However, populations of the similar R. maritima in tropical environments may suffer a decline as their productivity will be highly reduced. Full article

Berghia stephanieae is a stenophagous sea slug that preys upon glass anemones, such as Exaiptasia diaphana. Glass anemones host photosynthetic dinoflagellate endosymbionts that sea slugs ingest when consuming E. diaphana. However, the prevalence of these photosynthetic dinoflagellate endosymbionts in sea slugs appears to be short-lived, particularly if B.stephanieae is deprived of prey that host these microalgae (e.g., during bleaching events impacting glass anemones). In the present study, we investigated this scenario, along with food deprivation, and validated the use of a non-invasive and non-destructive approach employing chlorophyll fluorescence as a proxy to monitor the persistence of the association between sea slugs and endosymbiotic photosynthetic dinoflagellates acquired through the consumption of glass anemones. Berghia stephanieae deprived of a trophic source hosting photosynthetic dinoflagellate endosymbionts (e.g., through food deprivation or by feeding on bleached E. diaphana) showed a rapid decrease in minimum fluorescence (F_o) and photosynthetic efficiency (F_v/F_m) when compared to sea slugs fed with symbiotic anemones. A complete loss of endosymbionts was observed within 8 days, confirming that no true symbiotic association was established. The present work opens a new window of opportunity to rapidly monitor in vivo and over time the prevalence of associations between sea slugs and photosynthetic dinoflagellate endosymbionts, particularly during bleaching events that prevent sea slugs from incorporating new microalgae through trophic interactions. Full article

Environmental light conditions influence the biosynthesis of monoterpenes in the mint plant. Cyclic terpenes, such as menthol, menthone, pulegone, and menthofuran, are major odor components synthesized in mint leaves. However, it is unclear how light for cultivation affects the contents of these compounds. Artificial lighting using light-emitting diodes (LEDs) for plant cultivation has the advantage of preferential wavelength control. Here, we monitored monoterpene contents in hydroponically cultivated Japanese mint leaves under blue, red, or far-red wavelengths of LED light supplements. Volatile cyclic monoterpenes, pulegone, menthone, menthol, and menthofuran were quantified using the head-space solid phase microextraction method. As a result, all light wavelengths promoted the biosynthesis of the compounds. Remarkably, two weeks of blue-light supplement increased all compounds: pulegone (362% increase compared to the control), menthofuran (285%), menthone (223%), and menthol (389%). Red light slightly promoted pulegone (256%), menthofuran (178%), and menthol (197%). Interestingly, the accumulation of menthone (229%) or menthofuran (339%) was observed with far-red light treatment. The quantification of glandular trichomes density revealed that no increase under light supplement was confirmed. Blue light treatment even suppressed the glandular trichome formation. No promotion of photosynthesis was observed by pulse-amplitude-modulation (PAM) fluorometry. The present result indicates that light supplements directly promoted the biosynthetic pathways of cyclic monoterpenes. Full article

Recent studies suggest that photophysiological parameters for intact substrates with depth (e.g., periphytic biofilms, microphytobenthos) are overestimated by pulse-amplitude modulated (PAM) fluorometry. This overestimation results from depth-integration effects, following the activation of deeper photosynthesizing layers by an attenuated light signal. To mitigate this error, we propose a novel slide-based thin-film technique in which fluorescence is measured on a vertically representative subsample of the biofilm, spread evenly on a microscope slide. We compared bias and precision for photosynthetic parameters estimated through conventional PAM fluorometry on intact biofilms and through our novel slide-based technique, both theoretically and empirically. Numerical simulations confirmed the consistent overestimation of key parameters for intact biofilms, with relative errors up to 145%, compared to, at most, 52% on thin films. Paired empirical observations likewise demonstrated that estimates based on intact biofilms were consistently higher (up to 248%, $p<0.001$) than estimates from thin films. Numerical simulation suggested greater precision with the slide-based technique for homogeneous biofilms, but potentially less precision for heterogeneous biofilms with improper subsampling. Our empirical comparison, however, demonstrated some improvement in precision with the slide-based technique (e.g., the coefficient of variation for the maximum electron transport rate was reduced 30%, $p=0.009$). We recommend the use of the slide-based technique, particularly for biofilms that are thick or have small light attenuation coefficients. Care should be taken, however, to obtain vertically representative subsamples of the biofilm for measurement. Full article

Melting polar and alpine ice surfaces frequently exhibit blooms of dark pigmented algae. These microbial extremophiles significantly reduce the surface albedo of glaciers, thus accelerating melt rates. However, the ecology, physiology and taxonomy of cryoflora are not yet fully understood. Here, a Swiss and an Austrian glacier dominated either by filamentous Ancylonema nordenskioeldii or unicellular Mesotaenium berggrenii var. alaskanum, were sampled. Molecular analysis showed that both species are closely related, sharing identical chloroplast morphologies (parietal-lobed for Ancylonema vs. axial plate-like for Mesotaenium sensu stricto), thus the unicellular species was renamed Ancylonema alaskana. Moreover, an ecophysiological comparison of the two species was performed: pulse–amplitude modulated (PAM) fluorometry confirmed that they have a high tolerance to elevated solar irradiation, the physiological light preferences reflected the conditions in the original habitat; nonetheless, A. nordenskioeldii was adapted to higher irradiances while the photosystems of A. alaskana were able to use efficiently low irradiances. Additionally, the main vacuolar polyphenol, which effectively shields the photosystems, was identical in both species. Also, about half of the cellular fatty acids were polyunsaturated, and the lipidome profiles dominated by triacylglycerols were very similar. The results indicate that A. alaskana is physiologically very similar and closely related but genetically distinct to A. nordenskioeldii. Full article

Posidonia oceanica (L.) Delile meadows are recognized to be one of the most productive ecosystems of the Mediterranean basin. Due to the impacts of human activities in coastal areas, seagrasses are experiencing a critical decline. In this context, the understanding of the dynamics of production and photosynthesis in response to the environmental factors is essential to address efficient conservation strategies that limit this trend and to assess the ecological status of marine ecosystems. Pulse Amplitude Modulated (PAM) fluorometry has been widely implemented to assess seagrass health and productivity. Here we analyzed the photosynthetic dynamics of P. oceanica according to its bathymetric distribution and daily light availability along a depth gradient to be used as baseline for monitoring purposes on the health status of the seagrass meadows in the Northern Tyrrhenian Sea. Moreover, to investigate the effects of the environmental factors on the health status of P. oceanica within the study area through a multidisciplinary approach, the models contained in the Civitavecchia Coastal Environmental Monitoring System were used. In this study, significant photo-physiological changes have been observed among the investigated meadows. Moreover, the integration of physiological and hydrodynamic information allowed the description of how P. oceanica modulates its photosynthetic capacity at different environmental conditions. Full article

Plants naturally contain high levels of the stress-responsive fluorophore chlorophyll. Chlorophyll fluorescence imaging (CFI) is a powerful tool to measure photosynthetic efficiency in plants and provides the ability to detect damage from a range of biotic and abiotic stresses before visible symptoms occur. However, most CFI systems are complex, expensive systems that use pulse amplitude modulation (PAM) fluorometry. Here, we test a simple CFI system, that does not require PAM fluorometry, but instead simply images fluorescence emitted by plants. We used this technique to visualize stress induced by the photosystem II-inhibitory herbicide atrazine. After applying atrazine as a soil drench, CFI and color images were taken at 15-minute intervals, alongside measurements from a PAM fluorometer and a leaf reflectometer. Pixel intensity of the CFI images was negatively correlated with the quantum yield of photosystem II (ΦPSII) (p < 0.0001) and positively correlated with the measured reflectance in the spectral region of chlorophyll fluorescence emissions (p < 0.0001). A fluorescence-based stress index was developed using the reflectometer measurements based on wavelengths with the highest (741.2 nm) and lowest variability (548.9 nm) in response to atrazine damage. This index was correlated with ΦPSII (p < 0.0001). Low-cost CFI imaging can detect herbicide-induced stress (and likely other stressors) before there is visual damage. Full article

High-throughput and large-scale measurements of chlorophyll a fluorescence (ChlF) are of great interest to investigate the photosynthetic performance of plants in the field. Here, we tested the capability to rapidly, precisely, and simultaneously estimate the number of pulse-amplitude-modulation ChlF parameters commonly calculated from both dark- and light-adapted leaves (an operation which usually takes tens of minutes) from the reflectance of hyperspectral data collected on light-adapted leaves of date palm seedlings chronically exposed in a FACE facility to three ozone (O₃) concentrations (ambient air, AA; target 1.5 × AA O₃, named as moderate O₃, MO; target 2 × AA O₃, named as elevated O₃, EO) for 75 consecutive days. Leaf spectral measurements were paired with reference measurements of ChlF, and predictive spectral models were constructed using partial least squares regression. Most of the ChlF parameters were well predicted by spectroscopic models (average model goodness-of-fit for validation, R²: 0.53–0.82). Furthermore, comparing the full-range spectral profiles (i.e., 400–2400 nm), it was possible to distinguish with high accuracy (81% of success) plants exposed to the different O₃ concentrations, especially those exposed to EO from those exposed to MO and AA. This was possible even in the absence of visible foliar injury and using a moderately O₃-susceptible species like the date palm. The latter view is confirmed by the few variations of the ChlF parameters, that occurred only under EO. The results of the current study could be applied in several scientific fields, such as precision agriculture and plant phenotyping. Full article