Pulse-amplitude Modulated Fluorometer Research Articles

Effects of experimental CO2 enrichment on the PSII photochemical efficiency of Symbiodinium sp. in Acropora millepora

Enrichment of seawater with CO2 decreases the concentration of the carbonate ion while increasing that of hydrogen and bicarbonate ions. We use pulse-amplitude-modulation (PAM) fluorometry to investigate whether, in the absence of warming, and in sub-saturating light, these changes affect the PSII photochemical efficiency of _Symbiodinium_ sp. in the reef-building coral _Acropora millepora_. We assessed this experimentally with 30-min-interval saturation pulse analyses at 25 °C, a daily peak in the intensity of the photosynthetically active radiation (PAR) at ~65 µmol quanta m–2 s–1, and a seawater _p_CO2 that we gradually increased over nine days from ~496 to ~1290 μatm by injection of CO2-enriched air. Nine 14-day time series, which, except one, were recorded at the growing apices of a coral branch, revealed diel oscillations in the PSII photochemical efficiency characterized by a steep nocturnal decrease followed by a steep increase and peak in the morning, a daily minimum at midday (∆F/Fm’,midday), and a daily maximum at the onset of darkness at 19:00 h (Fv/Fm,19:00 h). An inadvertent shift in the position of one of the PAM fluorometer measuring heads revealed differences between the basal part and the growing coral apices of a coral branch in ∆F/Fm’midday and Qm. In ambient seawater (Control) _Symbiodinium_ sp. exhibited a gradual decrease, over the course of the experiment, in ∆F/Fm’,midday, Fv/Fm,19:00 h, and the slope of the linear regression between the relative electron transport rate and the intensity of PAR (rETR/PAR). Although two of three successive experiments indicated that CO2 enrichment counteracted these trends, statistical analyses failed to confirm an influence of _p_CO2 on ∆F/Fm’,midday, Fv/Fm,19:00 h, and Qm, rendering this experiment inconclusive.

Evaluation of selected tropical marine microalgal cultures for use in biophotovoltaic platforms

In this study, the bioelectrical power generation potential of four tropical marine microalgal strains native to Malaysia was investigated using BPV platforms. Chlorella UMACC 258 produced the highest power density (0.108 mW m−2), followed by Halamphora subtropica UMACC 370 (0.090 mW m−2), Synechococcus UMACC 371 (0.065 mW m−2) and Parachlorella UMACC 245 (0.017 mW m−2). The chlorophyll-a (chl-a) content was examined to have a linear positive relationship with the power density (p < 0.05). The photosynthetic performance of strains was studied using the pulse-amplitude modulation (PAM) fluorometer; parameters measured include the following: maximum quantum efficiency (Fv/Fm), alpha (α), maximum relative electron transport rate (rETRmax), photo-adaptive index (Ek) and non-photochemical quenching (NPQ). The Fv/Fm values of all strains, except Synechococcus UMACC 371, ranged between 0.37 and 0.50 during exponential and stationary growth phases, suggesting their general health during those periods. The low Fv/Fm value of Synechococcus UMACC 371 was possibly caused by the presence of background fluorescence from phycobilisomes or phycobiliproteins. Electrochemical studies via cyclic voltammetry (CV) suggest the presence of electrochemically active proteins on the cellular surface of strains on the carbon anode of the BPV platform, while morphological studies via field emission scanning electron microscope (FESEM) imaging verify the biocompatibility of the biofilms on the carbon anode.Key points• Maximum power output of 0.108 mW m−2 is recorded by Chlorella UMACC 258• There is a positive correlation between chl-a content and power output• Proven biocompatibility between biofilms and carbon anode sans exogenous mediators

The Photophysiology of Benthic Diatoms in the Intertidal Flats of Pulau Pinang (Malaysia)

The in-situ photosynthetic activity in tropical intertidal benthic diatom in response to environmental variation was assessed in this study by measuring chlorophyll fluorescence. The investigation was carried out during the lowest tide in January (non-rainy day) and February 2013 (post-rainy day) at two sampling sites (A and B) from each selected location (Pantai Jerejak, Teluk Bahang and Tanjung Bungah, Pulau Pinang, Malaysia). Samples of surface sediment (top 0.5 cm) were collected, and chlorophyll a extracted as biomass estimation. Assessments of the photosynthetic activity of benthic diatoms were made using a pulse-amplitude modulated (PAM) fluorometer. Fifty-three species were identified, representing 27 genera from the three studied locations. Both locations showed similarities in species diversity and abundance. Two-way ANOVA showed no significant differences (p = 0.430) in species richness (Margalef Index) among sampling locations, with an average value of 6.33±0.247. Both intertidal flats were dominated by Cocconeis, Navicula, Actinoptychus, and Diploneis. The community has low maximum quantum yields, Fv/Fm (ranging from 0.170 to 0.340) and is often light-limited (Photoacclimation Index, Ek, ranging from 67.96 to 236.71 mol photons m-2 s-1). The relative electron transport rate (rETRmax) was low, with values ranging from 3.45 to 35.51 across three sampling locations. Fluctuation in salinity has caused a decrease in photosynthetic activity. This study suggests that the low values indicated a poorly adapted benthic microalgal community that is constantly light-limited. However, time-series data is needed to determine the ability of these communities to adapt to the changing environment.

Concurrent bioimaging of microalgal photophysiology and oxidative stress.

The production of reactive oxygen species (ROS) is an unavoidable consequence of oxygenic photosynthesis and represents a major cause of oxidative stress in phototrophs, having detrimental effects on the photosynthetic apparatus, limiting cell growth, and productivity. Several methods have been developed for the quantification of cellular ROS, however, most are invasive, requiring the destruction of the sample. Here, we present a new methodology that allows the concurrent quantification of ROS and photosynthetic activity, using the fluorochrome dichlorofluorescein (DCF) and in vivo chlorophyll a fluorescence, respectively. Both types of fluorescence were measured using an imaging Pulse Amplitude Modulation (PAM) fluorometer, modified by adding a UVA-excitation light source (385nm) and a green bandpass emission filter (530nm) to enable the sequential capture of red chlorophyll fluorescence and green DCF fluorescence in the same sample. The method was established on Phaeodactylum tricornutum Bohlin, an important marine model diatom species, by determining protocol conditions that permitted the detection of ROS without impacting photosynthetic activity. The utility of the method was validated by quantifying the effects of two herbicides (DCMU and methyl viologen) on the photosynthetic activity and ROS production in P. tricornutum and of light acclimation state in Navicula cf. recens Lange-Bertalot, a common benthic diatom. The developed method is rapid and non-destructive, allowing for the high-throughput screening of multiple samples over time.

Protocol to control the invasive alga Avrainvillea lacerata in a shallow Hawaiian reef flat.

PremiseA novel control technique was developed to mitigate an invasive siphonous green alga, Avrainvillea lacerata (Dichotomosiphonaceae), within a shallow degraded reef flat in Oʻahu, Hawaiʻi.Methods and ResultsReplicated treatments of 3% and 10% hydrogen peroxide (H2O2) were administered into individual basal attachments of the bed‐forming invasive seaweed on the Paikō reef, Oʻahu. Relative electron transport rate maxima (rETRm) were measured using a Walz Diving Pulse Amplitude Modulated Fluorometer in two replicate 100‐m2 plots in 2020. Over the period of this short‐term study, rETRm decreased following injections of either concentration of H2O2 in contrast with negative and positive controls.ConclusionsCompared with existing techniques that have used oxidizing agents in the marine environment in localized areas, the protocol described here has the potential to successfully decrease macroalgal carbon gain, potentially leading to loss of biomass at larger scales.

Regulation of ppGpp Synthesis and Its Impact on Chloroplast Biogenesis during Early Leaf Development in Rice.

Guanosine tetraphosphate (ppGpp) is known as an alarmone that mediates bacterial stress responses. In plants, ppGpp is synthesized in chloroplasts from GTP and ATP and functions as a regulator of chloroplast gene expression to affect photosynthesis and plant growth. This observation indicates that ppGpp metabolism is closely related to chloroplast function, but the regulation of ppGpp and its role in chloroplast differentiation are not well understood. In rice, ppGpp directly inhibits plastidial guanylate kinase (GKpm), a key enzyme in GTP biosynthesis. GKpm is highly expressed during early leaf development in rice, and the GKpm-deficient mutant, virescent-2 (v2), develops chloroplast-deficient chlorotic leaves under low-temperature conditions. To examine the relationship between GTP synthesis and ppGpp homeostasis, we generated transgenic rice plants over-expressing RSH3, a protein known to act as a ppGpp synthase. When RSH3 was overexpressed in v2, the leaf chlorosis was more severe. Although the RSH3 overexpression in the wild type caused no visible effects, pulse amplitude modulation fluorometer measurements indicated that photosynthetic rates were reduced in this line. This finding implies that the regulation of ppGpp synthesis in rice is involved in the maintenance of the GTP pool required to regulate plastid gene expression during early chloroplast biogenesis. We further investigated changes in the expressions of RelA/SpoT Homolog (RSH) genes encoding ppGpp synthases and hydrolases during the same period. Comparing the expression of these genes with the cellular ppGpp content suggests that the basal ppGpp level is determined by the antagonistic action of multiple RSH enzymatic activities during early leaf development in rice.

Comparison of photosynthetic fluorescence characteristics of several submerged plants in Honghu Lake, China

Submerged plants are the pioneer species of eutrophic water remediation, and they are important for maintaining the health of aquatic ecosystem, while light is the main limiting factor for the growth of submerged plants. In this study, we measured the maximal quantum yields of photosystemⅡ(Fv/Fm) and rapid light curves (RLCs) of five dominant submerged macrophytes in situ by using pulse-amplitude modulated fluorometer (Diving-PAM). Results revealed that P. crispus L. and M. verticillatum L. had the highest Fv/Fm value, all species’ Fv/Fm are less than 0.8. In addition, the variation trends of Fv'/Fm' and Fv/Fm were same. All species showed statistically significant differences in α, while P. crispus L. and M. verticillatum L. showed the highest α value in the five species. And the variation trends of rETRm and Ek were basically the same. It indicated that P. crispus L. and M. verticillatum L., both of which had high photosynthetic efficiency, had excellent ability to withstand hard light. Compared five species, P. crispus L. and M. verticillatum L. had resistance capacity to hard light as well as faster photosynthetic rate, and V. natans (Lour.) Hara had higher resistance capacity to low light. Thus, when submerged plants are used for water restoration, V. natans (Lour.) Hara could be regarded as a pioneer species in eutrophication water restoration. P. crispus L. and M. verticillatum L. will have better effects when used in shallow water areas.

Environmental Stress Tolerance and Antioxidant Response of Palisada perforata (Rhodophyta) from a Tropical Reef1.

In this study, we analyzed the antioxidant activity and total phenolic content of the intertidal seaweed Palisada perforata collected from different reef microhabitats (sheltered site, tide pool, plateau, and exposed site) along the coast of Pernambuco (Brazil). Both parameters were compared with the same parameters of this species grown in the laboratory under two experiments simulating temperature, salinity, and desiccation conditions found in the reef. After both experiments (temperature x salinity and desiccation), the algal photosynthetic performance was measured through chlorophyll fluorescence parameters using a pulse-amplitude modulation fluorometer to test their stress response. Palisada perforata likely underwent stress by desiccation due to tidal fluctuations rather than to temperature or salinity changes. This conclusion agrees with our observations of the plateau site´s specimens, which were exposed to both air and UV radiation during low tides and exhibited higher antioxidant activity to avoid oxidative damage. However, despite the environmental stress, the antioxidant activity remained low, suggesting that photoinhibition is a crucial protection mechanism against oxidative damage.

Low-Cost Chlorophyll Fluorescence Imaging for Stress Detection

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.

Modification of a gas exchange system to measure active and passive chlorophyll fluorescence simultaneously under field conditions.

Solar-induced fluorescence (SIF) is a promising tool to estimate photosynthesis across scales; however, there has been limited research done at the leaf level to investigate the relationship between SIF and photosynthesis. To help bridge this gap, a LI-COR LI-6800 gas exchange instrument was modified with a visible-near-infrared (VIS-NIR) spectrometer to measure active and passive fluorescence simultaneously. The system was adapted by drilling a hole into the bottom plate of the leaf chamber and inserting a fibre-optic to measure passive steady-state fluorescence (F t , λ , analogous to SIF) from the abaxial surface of a leaf. This new modification can concurrently measure gas exchange, passive fluorescence and active fluorescence over the same leaf area and will allow researchers to measure leaf-level F t , λ in the field to validate tower-based and satellite measurements. To test the modified instrument, measurements were performed on leaves of well-watered and water-stressed walnut plants at three light levels and a constant air temperature. Measurements on these same plants were also conducted using a similarly modified Walz GFS-3000 gas exchange instrument to compare results. We found a positive linear correlation between F t , λ measurements from the modified LI-6800 and GFS-3000 instruments. We also report a positive linear relationship between F t , λ and normalized steady-state chlorophyll fluorescence (F t /F o ) from the pulse-amplitude modulation (PAM) fluorometer of the LI-6800 system. Accordingly, this modification will inform the link between spectrally resolved F t , λ and gas exchange-leading to improved interpretation of how remotely sensed SIF tracks changes in the light reactions of photosynthesis.

Ground Based Hyperspectral Imaging to Characterize Canopy-Level Photosynthetic Activities

Improving plant photosynthesis provides the best possibility for increasing crop yield potential, which is considered a crucial effort for global food security. Chlorophyll fluorescence is an important indicator for the study of plant photosynthesis. Previous studies have intensively examined the use of spectrometer, airborne, and spaceborne spectral data to retrieve solar induced fluorescence (SIF) for estimating gross primary productivity and carbon fixation. None of the methods, however, had a spatial resolution and a scanning throughput suitable for applications at the canopy and sub-canopy levels, thereby limiting photosynthesis analysis for breeding programs and genetics/genomics studies. The goal of this study was to develop a hyperspectral imaging approach to characterize plant photosynthesis at the canopy level. An experimental field was planted with two cotton cultivars that received two different treatments (control and herbicide treated), with each cultivar-treatment combination having eight replicate 10 m plots. A ground mobile sensing system (GPhenoVision) was configured with a hyperspectral module consisting of a spectrometer and a hyperspectral camera that covered the spectral range from 400 to 1000 nm with a spectral sampling resolution of 2 nm. The system acquired downwelling irradiance spectra from the spectrometer and reflected radiance spectral images from the hyperspectral camera. On the day after 24 h of the DCMU (3-(3,4-dichlorophenyl)-1,1-dimethylurea) application, the system was used to conduct six data collection trials in the experiment field from 08:00 to 18:00 with an interval of two hours. A data processing pipeline was developed to measure SIF using the irradiance and radiance spectral data. Diurnal SIF measurements were used to estimate the effective quantum yield and electron transport rate, deriving rapid light curves (RLCs) to characterize photosynthetic efficiency at the group and plot levels. Experimental results showed that the effective quantum yields estimated by the developed method highly correlated with those measured by a pulse amplitude modulation (PAM) fluorometer. In addition, RLC characteristics calculated using the developed method showed similar statistical trends with those derived using the PAM data. Both the RLC and PAM data agreed with destructive growth analyses. This suggests that the developed method can be used as an effective tool for future breeding programs and genetics/genomics studies to characterize plant photosynthesis at the canopy level.

Assessing plant performance in the Enviratron

BackgroundAssessing the impact of the environment on plant performance requires growing plants under controlled environmental conditions. Plant phenotypes are a product of genotype × environment (G × E), and the Enviratron at Iowa State University is a facility for testing under controlled conditions the effects of the environment on plant growth and development. Crop plants (including maize) can be grown to maturity in the Enviratron, and the performance of plants under different environmental conditions can be monitored 24 h per day, 7 days per week throughout the growth cycle.ResultsThe Enviratron is an array of custom-designed plant growth chambers that simulate different environmental conditions coupled with precise sensor-based phenotypic measurements carried out by a robotic rover. The rover has workflow instructions to periodically visit plants growing in the different chambers where it measures various growth and physiological parameters. The rover consists of an unmanned ground vehicle, an industrial robotic arm and an array of sensors including RGB, visible and near infrared (VNIR) hyperspectral, thermal, and time-of-flight (ToF) cameras, laser profilometer and pulse-amplitude modulated (PAM) fluorometer. The sensors are autonomously positioned for detecting leaves in the plant canopy, collecting various physiological measurements based on computer vision algorithms and planning motion via “eye-in-hand” movement control of the robotic arm. In particular, the automated leaf probing function that allows the precise placement of sensor probes on leaf surfaces presents a unique advantage of the Enviratron system over other types of plant phenotyping systems.ConclusionsThe Enviratron offers a new level of control over plant growth parameters and optimizes positioning and timing of sensor-based phenotypic measurements. Plant phenotypes in the Enviratron are measured in situ—in that the rover takes sensors to the plants rather than moving plants to the sensors.

Photosynthetic response and DNA mutation of tropical, temperate and polar Chlorella under short-term UVR stress

Changes in photosynthetic efficiency of Photosystem II can be used as an early stress indicator in phototrophs. In this study, chlorophyll fluorescence, measured by a Pulse-Amplitude Modulated Fluorometer (PAM), was used to determine the photosynthetic performance of tropical, temperate, Antarctic and Arctic Chlorella in response to short-term acute ultraviolet radiation (UVR) stress, with measurements of maximum quantum yield (Fv/Fm), photosynthetic efficiency (α), maximum electron transport rate (rETRm) and photoadaptive index (Ek). Three light treatments were conducted over a continuous, five-hour duration: (i) control subjected only to photosynthetic active radiation (PAR), (ii) PAR + UVA (UVA) and (iii) PAR + UVA + UVB (UVR). Tropical Chlorella showed better adaptive ability to UVA stress compared to strains from temperate and polar regions. UVB stress caused significant photosynthetic dysfunction in all samples, with polar strains showing a lower inhibition (about 40%) compared to the tropical strain (about 98%). Photosynthetic responses in Chlorella towards UVR are possibly origin dependent. DNA mutation induced by both UVA and UVR treatments was revealed by Random Amplified Polymorphic DNA (RAPD) analysis. Out of sixty RAPD primers tested, two primers: S33 (polymorphism degree 44.83%) and S90 (polymorphism degree 38.71%) were chosen as potential primers to conduct genomic study of UV stress in microalgae.

Photosynthetic fluorescence characteristics of six macroalgae species in seaweed beds of Gouqi Island, Zhejiang, China.

The study of photosynthetic fluorescence characteristics of algae is important for the analy-sis of photosynthesis and carbon sequestration of algae. In July 2017, six common species of macroalgae found in Gouqi seaweed beds were collected, including Ulva pertusa, Cladophora stimpsonii, Grateloupia livida, Sargassum thunbergii, Polysiphonia urceolata, and Hizikia fusifarme. In the field, the maximal quantum yieids of photosystem2(Fv/Fm) and rapid curves (RLCs) were mea-sured by using pulse-amplitude modulated fluorometer (Diving-PAM). The results showed that the measured maximal quantum yields of U. pertusa, C. stimpsonii, G. livida, S. thunbergii, P. urceolata, and H. fusiforme were 0.702, 0.704, 0.457, 0.618, 0.421 and 0.567, respectively. The Fv'/Fm' of six species were in order of C. stimpsonii>U. pertusa>S. thunbergii＞H. fusiforme＞G. livida＞P. urceolata. The difference between each species and significant difference was found in U. pertusa, C. stimpsonii, and H. fusiforme. H. fusiforme, S. thunbergii and U. pertusa had higher Pm and α than other species, indicating their higher photosynthetic capacity and better adaptation in higher light condition. However, G. livida had higher α but lower Ik, indicating G. livida had higher photosynthetic capacity in low light condition. In a word, differences of photosynthetic capacity and light intense tolerance between the three phyla of macroalgae were found and we suggested H. fusiforme, S. thunbergii and U. pertusa had stronger photosynthetic capacity and light intense tolerance. Our results could provide theoretical basis for the seaweed bed conservation and carbon sequestration of macroalgae.

Effect of Nutrient Loadings on the Regulation of Water Nitrogen and Phosphorus by Vallisneria natans and Its Photosynthetic Fluorescence Characteristics

Submerged macrophytes are an important component of aquatic ecosystems. During the growing period, submerged macrophytes can absorb nitrogen and phosphorus nutrients to reduce pollution loadings. Shoots of submerged macrophytes can also promote the adhesion of suspended substances in water, reducing the turbidity. The release of nutrients in sediments can be suppressed by its root system, and the resuspension of sediments caused by disturbance of winds and waves can also be resisted. The role of submerged macrophytes in ecological restoration of eutrophic lakes has attracted widespread attention. In 1960, the submerged plants Vallisneria natans and Potamogeton malaianus had been the dominant species in East Taihu. However after 2002, Nymphoides peltatum, Elodea nattalii, P. malaianus, etc. have gradually taken over the dominant roles along with significant elevations of nitrogen and phosphorus levels. Nutrients in water are not the only key factors causing eutrophication of water bodies; the nutrient source for submerged plant growth affect both the purification efficiency and the photosynthetic characteristics of submerged macrophytes. Excessive nitrogen and phosphorus concentrations can inhibit the photosynthetic physiological activities of submerged macrophytes, affecting the succession of aquatic vegetation. In addition, under high nutrient conditions, the competition from periphytic algae and planktonic algae may also directly poison submerged macrophytes, leading to its degradation and disappearance. Systematic studies on the regulation and photosynthetic fluorescence response mechanism of submerged macrophytes to varied nutrient loadings are helpful in revealing their relationships. The seedlings of submerged macrophyte V. natans were transplanted in a laboratory mesocosm to study the effect of nutrient loadings on its regulation of water nitrogen and phosphorus. Three nitrogen and phosphorus loadings from low, medium, and high levels derived from nitrate, ammonium, and phosphate were setup as the aquatic medium for the plant growth. Twelve harvests were carried out to determine the evolution of nutrient removal performance of V. natans. Its photosynthetic fluorescence characteristics were measured by a pulse-amplitude modulated fluorometer (Diving-PAM). Results showed that the nitrogen and phosphorus adsorption abilities of V. natans were gradually enhanced with the increase of nutrient concentrations in the range of TN ≤ 12 mg·L-1 and TP ≤ 1.0 mg·L-1. In the treatment of high nutrient concentrations (TN=12 mg·L-1 and TP=1.0 mg·L-1), the removal rates of nitrogen and phosphorus reached more than 95%. V. natans preferentially absorbed ammonium nitrogen when its concentration was high. The medium nutrient concentrations (TN:8-12 mg·L-1 and TP:0.6-1.0 mg·L-1) did not significantly affect the Fv/Fm ratio of leaves. However, the low nutrient concentrations (TN=3 mg·L-1 and TP=0.3 mg·L-1) could improve the Fv/Fm ratio of leaves and were beneficial for the growth of V. natans. The inhibition of photosynthetic activity and light tolerance were enhanced with the increase in nutrient concentrations. The photosynthetic activity of V. natans gradually recovered with no significant changes in the capacity for light harvesting, when the nutrient concentrations gradually decreased in the water. Our results indicate that the high nitrogen and phosphorus loadings indeed hamper the photosynthetic capacity, which may subsequently restrain the maintenance of the dominance of V. natans in the submerged macrophyte communities.

Photochemical Efficiencies of Diatom Symbionts in Hospite in Amphistegina Gibbosa (foraminifera) Across Seasons in the Florida Keys, Usa

Abstract Benthic foraminiferal taxa that host algal endosymbionts can serve as model organisms in understanding photo-oxidative stress in environments where algal symbioses are prevalent. This study examined photochemical performance of the diatom symbionts within the foraminifer Amphistegina gibbosa. Data on bleaching prevalence and photochemical efficiency for photosystem II (PSII) were obtained from specimens collected at Tennessee Reef, Florida reef tract, USA, as close as logistically feasible to each equinox and solstice in 2012 and 2013. Specimens were collected at 6 m and 18 m depths, isolated, visually characterized, and assessed for maximum quantum yield of PSII using a pulse-amplitude modulated (PAM) fluorometer. Rapid-light curves (RLCs) were performed in the laboratory on specimens within a few hours of collection. Trials examined the effects of depth, sampling date, and degree of host bleaching. Consistent with field studies in the 1990s, proportions of A. gibbosa populations exhibiting visible loss of symbionts (i.e., “bleaching”) were lowest in December, with between-season and between-year differences in maxima. Maximum quantum yields ranged from 0.66–0.76. Significant differences between sampling dates were found at the 6 m site, with highest yields in winter and lowest in summer. At the 18 m site, maximum quantum yields differed both by sampling date and by degree of bleaching. Rapid light curves revealed that relative electron transport rates were highest in specimens exhibiting no visible symbiont loss; derived-photosynthetic parameters of α (initial slope of the RLC) and Ek (minimum saturating irradiance) did not differ significantly across different degrees of symbiont loss. Overall results indicate that even individuals exhibiting substantial symbiont loss retain some fully functional symbionts.

Effect of elevated temperature on the physiological responses of marine Chlorella strains from different latitudes

The increased frequency of heat waves due to climate change poses a threat to all organisms. Microalgae are the basis of aquatic food webs, and high temperatures have significant impacts on their adaptation and survival rates. Algae respond to environmental changes by modulating their photosynthetic rates and biochemical composition. This study aims to examine the effect of elevated temperature on similar taxa of marine Chlorella originating from different latitudes. Strains from the Antarctic, temperate zone, and the tropics were grown at various temperatures, ranging from 4 to 38, 18 to 38, and 28 to 40 °C, respectively. A pulse-amplitude modulated (PAM) fluorometer was used to assess their photosynthetic responses. Parameters including maximum quantum efficiency (F v/F m), relative electron transport rate (rETR), and light harvesting efficiency (α) were determined from the rapid light curves (RLCs). In addition, the composition of fatty acids was compared to evaluate changes induced by the temperature treatments. Increasing the temperature from 35 to 38 °C for both Antarctic and temperate strains and from 38 to 40 °C for the tropical strain resulted in severe inhibition of photosynthesis and suppressed growth. Although all the strains demonstrated the ability to recover from different stress levels, the tropical strain was able to recover most rapidly while the Antarctic strain had the slowest recovery. The results underline that the thermal threshold for the analysed Chlorella strains temperature ranges between 38 and 40 °C. Furthermore, the analysed strains exhibited different trends in their response to elevated temperatures and recovery capabilities.

Chlorophyll fluorescence tracks seasonal variations of photosynthesis from leaf to canopy in a temperate forest.

Accurate estimation of terrestrial gross primary productivity (GPP) remains a challenge despite its importance in the global carbon cycle. Chlorophyll fluorescence (ChlF) has been recently adopted to understand photosynthesis and its response to the environment, particularly with remote sensing data. However, it remains unclear how ChlF and photosynthesis are linked at different spatial scales across the growing season. We examined seasonal relationships between ChlF and photosynthesis at the leaf, canopy, and ecosystem scales and explored how leaf-level ChlF was linked with canopy-scale solar-induced chlorophyll fluorescence (SIF) in a temperate deciduous forest at Harvard Forest, Massachusetts, USA. Our results show that ChlF captured the seasonal variations of photosynthesis with significant linear relationships between ChlF and photosynthesis across the growing season over different spatial scales (R2 =0.73, 0.77, and 0.86 at leaf, canopy, and satellite scales, respectively; P<0.0001). We developed a model to estimate GPP from the tower-based measurement of SIF and leaf-level ChlF parameters. The estimation of GPP from this model agreed well with flux tower observations of GPP (R2 =0.68; P<0.0001), demonstrating the potential of SIF for modeling GPP. At the leaf scale, we found that leaf Fq '/Fm ', the fraction of absorbed photons that are used for photochemistry for a light-adapted measurement from a pulse amplitude modulation fluorometer, was the best leaf fluorescence parameter to correlate with canopy SIF yield (SIF/APAR, R2 =0.79; P<0.0001). We also found that canopy SIF and SIF-derived GPP (GPPSIF ) were strongly correlated to leaf-level biochemistry and canopy structure, including chlorophyll content (R2 =0.65 for canopy GPPSIF and chlorophyll content; P<0.0001), leaf area index (LAI) (R2 =0.35 for canopy GPPSIF and LAI; P<0.0001), and normalized difference vegetation index (NDVI) (R2 =0.36 for canopy GPPSIF and NDVI; P<0.0001). Our results suggest that ChlF can be a powerful tool to track photosynthetic rates at leaf, canopy, and ecosystem scales.

Application of a pulse-amplitude-modulation (PAM) fluorometer reveals its usefulness and robustness in the prediction of Karenia mikimotoi blooms: A case study in Sasebo Bay, Nagasaki, Japan

Red tides of Karenia mikimotoi blooms have severely impacted marine fish culture in Japanese waters, however, blooms cannot be accurately predicted because they are controlled by a combination of numerous spontaneous environmental parameters, including unknown parameters. Nevertheless, these parameters must eventually control photosynthesis and result in growth, and measuring the photosynthetic activities of this species may be useful for predicting blooms. In this study, K. mikimotoi blooms were predicted in Sasebo Bay, Nagasaki, Japan, in 2014 and 2015 by measuring the maximum quantum yield (Fv/Fm) and relative photochemical electron transport rate (ETR) using pulse-amplitude-modulation (PAM) fluorometers (a cuvette-type and a microscopy-type fluorometer). In 2014, a raphidophycean Heterosigma akashiwo bloom initially dominated at the end of May and was followed by a K. mikimotoi bloom. This change was previously characterized by a decrease in the Fv/Fm of H. akashiwo and an increase in K. mikimotoi. In 2015, a moderate K. mikimotoi bloom was detected at the beginning of the survey and was predicted to increase thereafter based on the population’s high Fv/Fm and ETR values. Additionally, these values were highest in the mid-deeper layer, as expected for K. mikimotoi, which prefers lower light intensities in the water column. In both years, blooms of K. mikimotoi were replaced by Prorocentrum dentatum, and these successions were initially recognized by PAM fluorometer measurements. These dinoflagellate blooms eventually vanished due to massive diatom blooms, which were accelerated by nutrient inputs from rivers after heavy precipitation. Furthermore, the activation of these diatom populations was also primarily detected based on their Fv/Fm and ETR increases, especially in the surface layer. Again, this location is expected because diatoms prefer high light intensity. In this study, it was found that various environmental factors influenced the Fv/Fm and ETR values, especially light and nutrient stresses, and these values served as useful proxies for the phytoplankton physiological status. These values could be used to predict the blooming or succession of phytoplankton.

Toxicidade ambiental de efluentes advindo de diferentes laboratórios de uma farmácia magistral

Há uma crescente preocupação com os chamados “poluentes emergentes” na produção de medicamentos. Eles chegam ao meio ambiente a partir do descarte doméstico ou a partir das sobras geradas na produção de fármacos. Em pequenas concentrações da ordem de microgramas por litro ou inferiores, ou seja, concentrações não efetivas a curto prazo em termos de resposta biológica em seres humanos, podem causar grandes impactos no meio ambiente pela exposição crônica. O objetivo deste trabalho foi avaliar o risco de contaminação de efluentes pelo lançamento dos dejetos brutos da produção de medicamentos em pequena escala, por farmácias de manipulação, com base na avaliação da ecotoxicicidade pelo biomonitoramento de algas Euglena gracilis, e investigar os impactos apresentados sobre o comportamento e as alterações no processo de fotossíntese. Amostras de quatro laboratórios de diferentes demandas foram analisadas. As alterações comportamentais (velocidade de subida à superfície, r-value e velocidade de movimento) das algas foram biomonitoradas pelo NG-TOX e os parâmetros de fotossíntese foram medidos com o fluorímetro de pulso e amplitude modulada (PAM). Os resultados demonstraram que o efluente do laboratório de hormônios, que teve uma baixa produção semestral, apresentou pouco impacto, já o efluente do laboratório de psicotrópicos, mesmo com demanda intermediária, apresentou impacto significante sobre o comportamento e a atividade fotossintética das algas. A diferença de comportamento observada entre os diferentes setores de fármacos demonstra que os impactos e os possíveis riscos ambientais são diferentes para cada setor. A demanda e as diferentes substâncias manipuladas podem ser determinantes na classificação de risco e na escolha dos métodos de descontaminação.