Shining Light on Photosynthesis in the Harmful Dinoflagellate Karenia mikimotoi–Responses to Short-Term Changes in Temperature, Nitrogen Form, and Availability

Karenia mikimotoi is a toxic bloom-forming dinoflagellate that sometimes co-blooms with Karenia brevis in the Gulf of Mexico, especially on the West Florida Shelf where strong vertical temperature gradients and rapid changes in nitrogen (N) can be found. Here, the short-term interactions of temperature, N form, and availability on photosynthesis–irradiance responses were examined using rapid light curves and PAM fluorometry in order to understand their interactions, and how they may affect photosynthetic yields. Cultures of K. mikimotoi were enriched with either nitrate (NO3−), ammonium (NH4+), or urea with varying amounts (1, 5, 10, 20, 50 µM-N) and then incubated at temperatures of 15, 20, 25, 30 °C for 1 h. At 15–25 °C, fluorescence parameters (Fv/Fm, rETR) when averaged for all N treatments were comparable. Within a given light intensity, increasing all forms of N concentrations generally led to higher photosynthetic yields. Cells appeared to dynamically balance the “push” due to photon flux pressure and reductant generation, with consumption in overall metabolism (“pull” due to demand). However, at 30 °C, all fluorescence parameters declined precipitously, but differential responses were observed depending on N form. Cells enriched with urea at 30 °C showed a smaller decline in fluorescence parameters than cells treated with NO3− or NH4+, implying that urea might induce a photoprotective mechanism by increasing metabolic “pull”.

Photochemical changes during rehydration of an intertidal cyanobacterial mat exposed to variations in salinity and light intensity

This study focuses on a Lyngbya cf. aestuarii dominated mat community from the intertidal zone of the Laguna Ojo de Liebre, Baja California Sur. In this environment, the mat is desiccated for several days between spring tides. While the mats were desiccated, photosynthetic activity was absent but recovered rapidly (~3 h) upon rehydration. It has been shown previously that the rate of photosynthetic recovery is dependent on both light intensity and salinity. In the current study, photosynthetic recovery was measured based on chlorophyll a fluorescence using pulse amplitude modulated (PAM) fluorometry. Upon the addition of water, photosystem II (PSII) complexes recovered the capacity for reaction centre excitation. However, these functional centres were initially closed. Respiratory activity early in recovery probably reduced the plastoquinone pool through the shared use of part of the photosynthetic transport chain, thus temporarily blocking electron transport downstream of PSII. The time that PSII complexes remained closed increased with light intensities above saturation. This condition is potentially damaging to the cyanobacteria since the exposure of closed PSII centres to high light intensities can lead to the production of singlet oxygen. After this initial lag period, PSII centres opened rapidly indicating an increase in the flow of electrons from PSII to PSI. The rate of photosynthetic recovery appeared to be limited primarily by the relatively slow return of functional PSII. Photosynthetic recovery rates were slower in salinities greater than those that naturally occur in the intertidal zone.

A Novel Thin-Film Technique to Improve Accuracy of Fluorescence-Based Estimates for Periphytic Biofilms

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.

Unicellular versus Filamentous: The Glacial Alga Ancylonema alaskana comb. et stat. nov. and Its Ecophysiological Relatedness to Ancylonema nordenskioeldii (Zygnematophyceae, Streptophyta)

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.

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.