PSI Open Fluor CAM script for measuring qE component of NPQ in Chlorella vulgaris v1

This is a simple protocol that consists of 1) 10 minutes preillumination with far red light 2) 5 minutes of illumination with actinic light 3) 5 minutes of dark adaptation with far red light qE is calculated as the differe between NPQ_Lss and NPQ_D5 qE=NPQ_Lss-NPQ_D5 qI=NPQ_D5 Protocol to be used with FluorCAM 7.0 on a PSI Open FC 800-O/1010-S. Act 2 - are the white light LED arrays ADD2 - is the far red LED array Camera is placed at ~20 cm above the measured sample. Light intensity uniformity across the 96 well plate was measured according to manufacturer instructions. !Important - protocol only works under weak far red light. Intense far red will interfere with the fluorescence measurement.

Elevated CO2 Concentration Alters Photosynthetic Performances under Fluctuating Light in Arabidopsis thaliana

In view of the current and expected future rise in atmospheric CO2 concentrations, we examined the effect of elevated CO2 on photoinhibition of photosystem I (PSI) under fluctuating light in Arabidopsis thaliana. At 400 ppm CO2, PSI showed a transient over-reduction within the first 30 s after transition from dark to actinic light. Under the same CO2 conditions, PSI was highly reduced after a transition from low to high light for 20 s. However, such PSI over-reduction greatly decreased when measured in 800 ppm CO2, indicating that elevated atmospheric CO2 facilitates the rapid oxidation of PSI under fluctuating light. Furthermore, after fluctuating light treatment, residual PSI activity was significantly higher in 800 ppm CO2 than in 400 ppm CO2, suggesting that elevated atmospheric CO2 mitigates PSI photoinhibition under fluctuating light. We further demonstrate that elevated CO2 does not affect PSI activity under fluctuating light via changes in non-photochemical quenching or cyclic electron transport, but rather from a rapid electron sink driven by CO2 fixation. Therefore, elevated CO2 mitigates PSI photoinhibition under fluctuating light at the acceptor rather than the donor side. Taken together, these observations indicate that elevated atmospheric CO2 can have large effects on thylakoid reactions under fluctuating light.

Evidence for variable chlorophyll fluorescence of photosystem I in vivo

Room temperature fluorescence in vivo and its light-induced changes are dominated by chlorophyll a fluorescence excited in photosystem II, F(II), peaking around 685 nm. Photosystem I fluorescence, F(I), peaking around 730 nm, so far has been assumed to be constant in vivo. Here, we present evidence for significant contributions of F(I) to variable fluorescence in the green unicellular alga Chlorella vulgaris, the cyanobacterium Synechococcus leopoliensis and a light-green ivy leaf. A Multi-Color-PAM fluorometer was applied for measurements of the polyphasic fluorescence rise (O-I1-I2-P) induced by strong 440 nm light in a dilute suspension of Chlorella, with detection alternating between emission above 700 nm (F > 700) and below 710 nm (F < 710). By averaging 10 curves each of the F > 700 and F < 710 recordings even small differences could be reliably evaluated. After equalizing the amplitudes of the O-I1 phase, which constitutes a specific F(II) response, the O-I1-I2 parts of the two recordings were close to identical, whereas the I2-P phase was larger in F > 700 than in F < 710 by a factor of 1.42. In analogous measurements with Synechococcus carried out in the dark state 2 using strong 625 nm actinic light, after O-I1 equalization the I2-P phase in F > 700 exceeded that in F < 710 even by a factor of 1.99. In measurements with Chlorella, the I2-P phase and with it the apparent variable fluorescence of PS I, Fv(I), were suppressed by moderate actinic background light and by the plastoquinone antagonist DBMIB. Analogous measurements with leaves are rendered problematic by unavoidable light intensity gradients and the resulting heterogenic origins of F > 700 and F < 710. However, a light-green young ivy leaf gave qualitatively similar results as those obtained with the suspensions, thus strongly suggesting the existence of Fv(I) also in leaves.

Photo-tunable epsilon-near-zero behavior in a self-assembled liquid crystal – nanoparticle hybrid material

A room temperature stable self-assembled liquid crystal metamaterial exhibits dynamic tuning of the epsilon-near-zero (ENZ) bandwidth on illumination with actinic light in the optical regime.

LHCSR3-Type NPQ Prevents Photoinhibition and Slowed Growth under Fluctuating Light in Chlamydomonas reinhardtii

Natural light intensities can rise several orders of magnitude over subsecond time spans, posing a major challenge for photosynthesis. Fluctuating light tolerance in the green alga Chlamydomonas reinhardtii requires alternative electron pathways, but the role of nonphotochemical quenching (NPQ) is not known. Here, fluctuating light (10 min actinic light followed by 10 min darkness) led to significant increase in NPQ/qE-related proteins, LHCSR1 and LHCSR3, relative to constant light of the same subsaturating or saturating intensity. Elevated levels of LHCSR1/3 increased the ability of cells to safely dissipate excess light energy to heat (i.e., qE-type NPQ) during dark to light transition, as measured with chlorophyll fluorescence. The low qE phenotype of the npq4 mutant, which is unable to produce LHCSR3, was abolished under fluctuating light, showing that LHCSR1 alone enables very high levels of qE. Photosystem (PS) levels were also affected by light treatments; constant light led to lower PsbA levels and Fv/Fm values, while fluctuating light led to lower PsaA and maximum P700+ levels, indicating that constant and fluctuating light induced PSII and PSI photoinhibition, respectively. Under fluctuating light, npq4 suffered more PSI photoinhibition and significantly slower growth rates than parental wild type, whereas npq1 and npq2 mutants affected in xanthophyll carotenoid compositions had identical growth under fluctuating and constant light. Overall, LHCSR3 rather than total qE capacity or zeaxanthin is shown to be important in C. reinhardtii in tolerating fluctuating light, potentially via preventing PSI photoinhibition.

Chloroplast-derived photo-oxidative stress causes changes in H2O2 and EGSH in other subcellular compartments

ABSTRACTMetabolic fluctuations in chloroplasts and mitochondria can trigger retrograde signals to modify nuclear gene expression. Mobile signals likely to be involved are reactive oxygen species (ROS), which can operate protein redox switches by oxidation of specific cysteine residues. Redox buffers such as the highly reduced glutathione pool serve as reservoirs of reducing power for several ROS scavenging and ROS-induced damage repair pathways. Formation of glutathione disulfide (GSSG) and a shift of the glutathione redox potential (EGSH) towards less negative values is considered a hallmark of several stress conditions. Here we used the herbicide methyl viologen (MV) to generate ROS locally in chloroplasts of intact Arabidopsis seedlings and recorded dynamic changes in EGSH and H2O2 levels with the genetically-encoded biosensors Grx1-roGFP2 (for EGSH) and roGFP2-Orp1 (for H2O2) targeted to chloroplasts, the cytosol or mitochondria. Treatment of seedlings with MV caused a rapid oxidation in chloroplasts and subsequently also in the cytosol and mitochondria. The MV-induced oxidation was significantly boosted by illumination with actinic light and largely abolished by inhibitors of photosynthetic electron transport. In addition, MV also induced an autonomous oxidation in the mitochondrial matrix in an electron transport chain activity-dependent manner that was milder than the oxidation triggered in chloroplasts by the combination of MV and light. In vivo redox biosensing resolves the spatiotemporal dynamics of compartmental responses to local ROS generation and provide a basis for understanding how compartment-specific redox dynamics may operate in retrograde signaling and stress acclimation in plants.One sentence summaryMethyl viologen-induced photooxidative stress causes an increase of H2O2 and oxidation of glutathione in chloroplasts, cytosol and mitochondria as well as autonomous oxidation in mitochondria.

Recommendations for spectral fitting of SO₂ from MAX-DOAS measurements

Fitting SO2 dSCDs from MAX-DOAS measurements of scattered sunlight is challenging because actinic light intensity is low in wavelength regions where the SO2 absorption features are strongest. SO2 dSCDs were fit with different wavelength windows (λlow to λhigh) from ambient measurements with calibration cells of 2.2 × 1017 and 2.2 × 1016 molec cm−2 inserted in the light path at different viewing elevation angles. SO2 dSCDs were the least accurate and fit errors were the largest for fitting windows with λlow 312 nm. The SO2 dSCDs also exhibited an inverse relationship with the SO2 absorption cross-section for fitting windows with λlow 400 nm. Deviation of the SO2 dSCD from the true value depended on the SO2 concentration for some fitting windows rather than exhibiting a consistent bias. Uncertainties of the SO2 dSCD reported by the fit algorithm were significantly less than the true error for many windows, particularly for the measurements without the filter or offset function. For retrievals with the filter or offset function, increasing λhigh > 320 nm tended to decrease the reported fit uncertainty but did not increase the accuracy. Based on the results of this study, a short-pass filter and a fitting window of 307.5

Analysis of Light-Induced Changes in the Photochemical Reflectance Index (PRI) in Leaves of Pea, Wheat, and Pumpkin Using Pulses of Green-Yellow Measuring Light

The photochemical reflectance index (PRI) is a widely used spectral index which can show stress-induced changes in photosynthesis (e.g., increase of the nonphotochemical quenching of chlorophyll fluorescence (NPQ)). The artificial illumination of plants improves the efficiency of estimation of photosynthetic processes on the basis of PRI measurements. However, the simultaneous activity of different light sources with different locations can disturb the measurement of PRI. Using pulses of a green-yellow measuring light can potentially solve this problem. The aim of the present work was to investigate the possibility of using green-yellow light pulses for the investigation of light-induced changes in PRI in higher plants (pea, wheat, and pumpkin) and for the analysis of connection between PRI and the energy-dependent component of NPQ (NPQF). First, we showed that using green-yellow light pulses eliminated shifts of reflected light, which were connected with the application of a red actinic light. Second, analysis of light dependences of NPQF, the absolute value of PRI, and changes in PRI (the difference between the PRI under the actinic light and the initial value of PRI without this light, ΔPRI) showed that the dynamics of the increase of NPQF and the decrease of PRI and ΔPRI were similar. Changes in NPQF and ΔPRI were found to be significant. In contrast, changes in the absolute value of PRI were not significant in most of the variants of the experiments. Third, scatter plots between NPQF and ΔPRI showed similar linear correlations for investigated species; moreover, a total set of experimental points (for pea, wheat, and pumpkin) were also described by the same linear regression. Thus, our results show that (i) pulses of green-yellow measuring light can be used for measurements of PRI, and (ii) ΔPRI is a more effective indicator for the estimation of NPQ than the absolute value of PRI.

Digital Luminescence Patterning via Inkjet Printing of a Photoacid Catalysed Organic-Inorganic Hybrid Formulation

Accurate positioning of luminescent materials at the microscale is essential for the further development of diverse application fields including optoelectronics, energy, biotechnology and anti-counterfeiting. In this respect, inkjet printing has recently attracted great interest due to its ability to precisely deposit with high throughput and no contact, functional materials on different types of substrates. Here, we present a novel photoacid catalysed organic-inorganic hybrid luminescent ink. The formulation, containing monomers bearing epoxy and silane functionalities, a photoacid generator and a small percentage of Rhodamine-B, shows good jetting properties and adequate wetting of the deposited droplets on the receiving substrates. Ultraviolet exposure of the deposited material triggers the cationic ring-opening polymerization reaction of the epoxy groups. Concomitantly, if atmospheric water is available, hydrolysis and condensation takes place, overall leading to a luminescent crosslinked hybrid organic-inorganic polymeric material obtained through a simple one-step curing process, without post baking steps. Advantageously, protection of the ink from actinic light delays the hydrolysis and condensation conferring long-term stability to the ink. Digital patterning leads to patterned emissive surfaces and elements with good adhesion to different substrates, mechanical and optical properties for the fabrication of optical and photonic elements and devices.