Fluorescence Measurements Research Articles

Dynamic responses of chlorophyll fluorescence parameters to drought across diverse plant families.

Chlorophyll fluorescence measurement is a quick and efficient tool for plant stress-level detection. The use of Pulse amplitude modulation (PAM), allows the detection of the plant stress level under field conditions. Over the years, several parameters estimating different parts of the chlorophyll and photosystem response were developed to describe the plant stress level. Despite all fluorescence parameters being based on the same measurements, their relationship remains unclear, and their response to drought stress is significantly influenced by the incoming light intensity. In this study, we use six different annual plants from different families, both C3 and C4 photosynthesis types, to describe the plant response to drought through the fluorescence parameters response (NPQ, Y(NPQ), and qN). To describe the dynamic response to drought, we employed light-response curves, adapting and fitting an equation for each curve to compare the drought response for each fluorescence parameter. The results demonstrated that the non-photochemical quenching (NPQ) and the quantum yield of non-photochemical quenching [Y(NPQ)] maximal values decrease when the PSII functionality (Fv/Fm) is lower than ~0.7. The basal fluorescence level ( and remained unaffected by the stress level and stayed stable across the various plants and stress levels. Our results indicate that the response of different stress parameters follows a distinct order under continuous drought. Consequently, monitoring just one parameter during long-term stress assessments may result in biased analysis outcomes. Incorporating multiple chlorophyll fluorescence parameters offers a more accurate reflection of the plant's stress level.

Single-molecule force spectroscopy of toehold-mediated strand displacement

Toehold-mediated strand displacement (TMSD) is extensively utilized in dynamic DNA nanotechnology and for a wide range of DNA or RNA-based reaction circuits. Investigation of TMSD kinetics typically relies on bulk fluorescence measurements providing effective, bulk-averaged reaction rates. Information on individual molecules or even base pairs is scarce. In this work, we explore the dynamics of strand displacement processes at the single-molecule level using single-molecule force spectroscopy with a microfluidics-enhanced optical trap supported by state-of-the-art coarse-grained simulations. By applying force, we can trigger and observe TMSD in real-time with microsecond and nanometer resolution. We find TMSD proceeds very rapidly under load with single step times of 1 µs. Tuning invasion efficiency by introducing mismatches allows studying thousands of forward/backward invasion events on a single molecule and analyze the kinetics of the invasion process. Extrapolation to zero force reveals single step times for DNA invading DNA four times faster than for RNA invading RNA. We also study the kinetics of DNA invading RNA, a process that in the absence of force would rarely occur. Our results reveal the importance of sequence effects for the TMSD process and have relevance for a wide range of applications in nucleic acid nanotechnology and synthetic biology.

An europium complex assembled by a flexible derivative of imidazole-4,5-dicarboxylic acid as a fluorescence turn-off chemosensor to detect Cr(VI) anions

A new europium(III) CP [Eu(HBBIDA)(H2O)]n (1) (H4BBIDA = 1,1′-(butane-1,4-diyl)bis(1H-imidazole-4,5-dicarboxylic acid) was successfully constructed by hydrothermal synthesis method and detailed characterization was performed. Crystal structure analysis reveals that CP 1 presents a 2D layered structure composed of cage-like SBUs with a TiS2 topological network. CP 1 can maintain framework integrity in an air environment below 260 °C, as well as in a certain pH range of acid-base aqueous solutions and common organic reagents, showing better thermal and chemical stability. Fluorescence measurements show that CP 1 exhibited excellent sensing activities toward CrO42− and Cr2O72− anions through fluorescence quenching, and their average quenching Ksv values of 3.99 × 103 M−1 (CrO42−) and 4.36 × 103 M−1 (Cr2O72−) and the LOD of 3.30 × 10−6 M (CrO42−) and 3.02 × 10−6 M, which makes it very suitable for the detection of hexavalent chromium anions contaminants in aqueous solutions. Furthermore, possible sensing mechanisms have been studied and discussed in detail, and the results show that static photocompetitive absorption should be the primary cause for the fluorescence quenching of the sensor.

Metal-based compounds: Synthesis and characterization of new thiazole-based iridium and palladium complexes with potential anticancer and other biological activities

Thiazoles and their derivatives are one of the most active classes of compounds known for their wide spectrum of bioactivity. Metal complexes, based on them, show antitumor potential that is attractive for investigations. Herein, we report 6 new biologically active thiazole-based complexes have been synthesized. The iridium- and palladium-based coordination compounds obtained by the precipitation method were characterized using elemental analysis (EA), Fourier-transform infrared spectroscopy (FTIR), magnetic measurements, thermogravimetric analysis coupled with mass spectrometry (TGA-MS), and scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX). Spectroscopic data helped to propose the formulas of the complexes and indicated that all ligands acted in a monodentate manner. Water molecules were identified by thermal analysis and FTIR spectroscopy. Mathematical analysis and evaluation of thermodynamic parameters including entropy (ΔS), Gibbs free energy (ΔG), and activation energy (E) were performed using the Coats–Redfern method for all complexes. The biological potential (anticancer, antibacterial, and antifungal properties) of compounds was analyzed by biological evaluation studies. Investigated CT-DNA studies revealed that the prepared compounds were intercalatively bound to the DNA. Cytotoxicity analyses showed that complexation with Ir(III) increased the toxicity of L2 towards both tested cell lines (LN-229 and MDA-MB-231), while complexation of L3 with Pd(II) significantly increased cytotoxic activity against LN-229. Due to this, the further biological studies, such as apoptosis/necrosis detection, cell cycle analysis and JC-1 fluorescence measurements were performed on this pair of compounds.

Evaluation of Novel Diaza Cage Compounds as MRP Modulators in Cancer Cells.

Novel MRP modulators are needed to combat MRP-mediated multidrug resistance (MDR) in cancer cells. Anticancer drug resistance is the main problem in cancer therapy. Causative multidrug efflux pumps are attractive target structures for the development of inhibitors of their activity. We synthesized novel cage dimeric 1,4-dihydropyridines to evaluate them as MRP modulators in cancer cells targeting MRP1, MRP2, and MRP4. Cage compounds were synthesized by solution dimerization of monomeric 1,4-dihydropyridines and a final functionalization reaction. The MRP modulation was determined in cellular efflux assays by the use of the flow cytometry technique as well as cellular fluorescent measurements with each fluorescent substrate of the efflux pumps. Difluoro phenyl and methoxy or dimethoxy benzyl substitutions were most favourable for the MRP1 and MRP2 inhibition, whereas monofluor phenyl and dimethoxy benzyl substitutions were most favourable for the MRP4 inhibition. Effective inhibitors were identified that were demonstrated to restore the respective cancer cell line sensitivity for the anticancer drug as a proof-of-concept that encourages further preclinical studies.</p>.

Impedance‐based monitoring of titration and neutralization assays with VSV‐G and SARS‐CoV‐2‐spike pseudoviruses

AbstractSince cell‐based virus neutralization assays are still the gold standard to assess a patient's immune protection against a given virus, they are of utmost importance for serodiagnosis, convalescent plasma therapy, and vaccine development. Monitoring the emergence and characteristics of neutralizing antibodies in an outbreak situation, confirming neutralizing antibodies as correlates of protection from infection and testing vaccine‐induced potency of neutralizing antibody responses, quests for automated, fast, and parallel neutralization assays. We developed an impedance‐based sensor platform (electric cell‐substrate impedance sensing, ECIS) providing time‐resolved monitoring of the host cell response to viral pseudotypes. For validation, the impedance assay was compared with state‐of‐the‐art quantification of virus‐induced reporter protein expression as an independent indicator of virus infection and neutralization. Vesicular stomatitis virus (VSV) derived pseudoviruses encoding the green fluorescent protein (GFP) as reporter and the autologous G protein (VSV‐G) for the initial binding to the host cell membrane were used for monitoring of HEK293T cell infection and neutralization with both, impedance and optical readout. Virus‐induced cytopathic effects (CPE) were detectable for low pseudotype concentrations (multiplicity of infection 1) in time‐resolved impedance profiles as soon as 5–10 h after infection in a concentration‐dependent manner. Neutralization efficacy of α‐VSV‐G antibodies was determined from impedance time courses and IC50 values compared favorably with fluorescence measurements of virus‐borne GFP expression. Sera of convalescent COVID‐19 patients were tested successfully for SARS‐CoV‐2 neutralizing antibodies by incubating VSV, pseudotyped with the SARS‐CoV‐2 spike protein, with different sera before host cell exposure and impedance recordings. In summary: (i) ECIS monitoring was successfully applied to detect virus‐mediated cell infection and neutralization; (ii) Impedance‐based monitoring allows reducing the assay time to 5–10 h; and (iii) the platform is easily adapted to other virus‐based diseases and scalable to high‐throughput.

Continuous-Flow Synthesis of Zn1-xMnxS Nanoparticles at Ambient Conditions.

With its large direct band gap and good chemical stability, ZnS is suitable for many applications, including light-emitting diodes, panel displays, and photodetection. Here, nanoparticles of ZnS are synthesized phase pure under ambient conditions by precipitation in a simple and scalable continuous-flow reactor. Furthermore, different degrees of Zn substitution with Mn have been investigated, Zn1-xMnxS, with x = 0.05, 0.19, and 0.25 according to X-ray fluorescence measurements. The products are analyzed with multitemperature synchrotron powder X-ray diffraction (PXRD) and X-ray total scattering. The analysis reveals phase-pure synthesis products with the sphalerite structure and crystallite sizes in the range of 3.8-4.7 nm in agreement with scanning transmission electron microscopy. Only Zn0.75Mn0.25S shows traces of Mn3O4, indicating that x = 0.25 is above the substitution limit as the impurity appears. Substitution of Zn with Mn in the nanoparticles is confirmed by energy-dispersive X-ray spectroscopy, as well as an observed decrease in the band gap, decrease in the sphalerite-to-wurtzite phase transition temperature, and increase in the unit cell dimensions with increasing Mn content. Based on the modeling of the PXRD Rietveld refined atomic displacement parameters, the Debye temperature for ZnS and Zn0.95Mn0.05S is determined to be 322 ± 13 and 394 ± 22 K, respectively.

Chemical speciation of methylmercury and mercury(II) cations in fish by new fluorogenic naphthalimide alkynyl gold complexes: The ultimate test for detecting fish contamination

The proof of concept of a new portable protocol for the detection and speciation of methylmercury and mercury(II) cations in fish samples is described. In this way, traces of methylmercury and/or mercury(II) cations in fish were quantitatively detected using an elaborated protocol and including a fluorescent sensor. In-the-field accurate results were obtained due to the selective behavior of the chemical sensor in the presence of both cations, working at very low concentrations of mercury species in fish under standard laboratory conditions, that are suitable for daily use in ordinary sampling by border companies importing fish or remote laboratories where large devices cannot be carried. The reported fish protocol is useful under real-life conditions for analyzing fish for the presence of methylmercury in contrast to total mercury analysis. The combination of the protocol and the fluorescent reporter constitutes the ultimate test for fish contamination by mercury(II) species in the workplace. The reported limits of detection of 0.72 nM for Hg2+ and 1.62 nM for CH3Hg+ and the closely related values of the two mercury(II) species in fish samples, obtained by ICP-MS analysis and fluorescent measurements, make the method suitable for daily control of methylmercury in fish under the EU legislation parameters and the WHO recommendations.

Substituent effects on first generation photochemical molecular motors probed by femtosecond stimulated Raman.

Unidirectional photochemical molecular motors can act as a power source for molecular machines. The motors operate by successive excited state isomerization and ground state helix inversion reactions, attaining unidirectionality from an interplay of steric strain and stereochemistry. Optimizing the yield of the excited state isomerization reaction is an important goal that requires detailed knowledge of excited state dynamics. Here, we investigate the effect of electron withdrawing and donating substituents on excited state structure and ultrafast dynamics in a series of newly synthesized first generation photochemical molecular motors. All substituents red-shift the absorption spectra, while some modify the Stokes shift and render the fluorescence quantum yield solvent polarity dependent. Raman spectra and density functional theory calculations reveal that the stretching mode of the C=C "axle" in the electronic ground state shows a small red-shift when conjugated with electron withdrawing substituents. Ultrafast fluorescence measurements reveal substituent and solvent polarity effects, with the excited state decay being accelerated by both polar solvent environment and electron withdrawing substituents. Excited state structural dynamics are investigated by fluorescence coherence spectroscopy and femtosecond stimulated Raman spectroscopy. The time resolved Raman measurements are shown to provide structural data specifically on the Franck-Condon excited state. The C=C localized modes have a different substituent dependence compared to the ground state, with the unsubstituted motor having the most red-shifted mode. Such measurements provide valuable new insights into pathways to optimize photochemical molecular motor performance, especially if they can be coupled with high-quality quantum molecular dynamics calculations.

In-Situ Generation of Hydrogen Polysulfides (H2Sn) with Thioglucose, Glucose Oxidase, and Chloroperoxidase.

Hydrogen polysulfides (H2Sn) have emerged as critical physiological mediators that are closely associated with hydrogen sulfide (H2S) signaling. H2Sn exhibit greater nucleophilicity than H2S while also having electrophilic characteristics, enabling unique activities such as protein S-persulfidation. Despite their physiological importance, mechanisms and reactivities of H2Sn remain inadequately explored due to their inherent instability in aqueous environments. Consequently, there is a need to develop biocompatible methods for controlled H2Sn generation to elucidate their behaviors in biological contexts. Herein, we present a dual enzyme system (containing glucose oxidase (GOx) and chloroperoxidase (CPO)) with thioglucose as the substrate to facilitate the controlled release of H2Sn. Fluorescence measurements with SSP4 and the trapping studies allowed us to confirm the production of H2Sn. Such a method may be useful in elucidating the reactivity of hydrogen polysulfides in biological systems as well as provide a potential delivery of H2Sn to target sites for biological applications.

Seasonal timing of fluorescence and photosynthetic yields at needle and canopy scales in evergreen needleleaf forests.

The seasonal timing and magnitude of photosynthesis in evergreen needleleaf forests (ENFs) has major implications for the carbon cycle and is increasingly sensitive to changing climate. Earlier spring photosynthesis can increase carbon uptake over the growing season or cause early water reserve depletion that leads to premature cessation and increased carbon loss. Determining the start and the end of the growing season in ENFs is challenging due to a lack of field measurements and difficulty in interpreting satellite data, which are impacted by snow and cloud cover, and the pervasive "greenness" of these systems. We combine continuous needle-scale chlorophyll fluorescence measurements with tower-based remote sensing and gross primary productivity (GPP) estimates at three ENF sites across a latitudinal gradient (Colorado, Saskatchewan, Alaska) to link physiological changes with remote sensing signals during transition seasons. We derive a theoretical framework for observations of solar-induced chlorophyll fluorescence (SIF) and solar intensity-normalized SIF (SIFrelative) under snow-covered conditions, and show decreased sensitivity compared with reflectance data (~20% reduction in measured SIF vs. ~60% reduction in near-infrared vegetation index [NIRv] under 50% snow cover). Needle-scale fluorescence and photochemistry strongly correlated (r2 = 0.74 in Colorado, 0.70 in Alaska) and showed good agreement on the timing and magnitude of seasonal transitions. We demonstrate that this can be scaled to the site level with tower-based estimates of LUEP and SIFrelative which were well correlated across all sites (r2 = 0.70 in Colorado, 0.53 in Saskatchewan, 0.49 in Alaska). These independent, temporally continuous datasets confirm an increase in physiological activity prior to snowmelt across all three evergreen forests. This suggests that data-driven and process-based carbon cycle models which assume negligible physiological activity prior to snowmelt are inherently flawed, and underscores the utility of SIF data for tracking phenological events. Our research probes the spectral biology of evergreen forests and highlights spectral methods that can be applied in other ecosystems.

Integrated Active Quenching Circuit for high-rate and distortionless SPAD-based time-resolved fluorescence applications.

Time-Correlated Single Photon Counting (TCSPC) is a pivotal technique in low-light-detection applications, renowned for its exceptional sensitivity and bandwidth, widely used in Fluorescence Lifetime Imaging Microscopy (FLIM) and quantum optics. Despite its features, TCSPC is significantly hindered by the pile-up effect, which may distort measurements at high photon-detection rates. Overcoming pile-up is challenging, with traditional solutions often involving complex post-processing or multichannel systems, complicating the TCSPC setup and limiting performance. A breakthrough to overcome this issue is matching the photodetector dead time to an integer multiple of the laser period, obtaining a distortionless histogram even at high illumination conditions. Building on this concept, we present an Active Quenching Circuit (AQC) developed in high-voltage 150 nm technology, achieving unprecedented control over the Single Photon Avalanche Diode (SPAD) dead time. Our design compensates for Process, Voltage, and Temperature (PVT) variations, ensuring ultra precise and robust dead time tuning. The presented AQC achieves a dead-time resolution of 50 ps suitable for time-resolved experiments within a selectable range of laser frequencies from 20 to 100 MHz, maintaining close-to- ideal linearity in dead-time control. Experimental validations through fluorescence measurements reveal a distortion as low as 0.43% under elevated count-rate conditions, highlighting the efficacy of our circuit in overcoming the pile-up limitation.

Research on airfoil surface flow structure testing based on Tuft velocity measurement method

Fluorescent tuft visualization technology, as a practical and convenient technique, has attracted widespread attention. In this study, a novel fluorescent tuft velocity measurement method is developed based on existing technologies. This method involves the recognition of tuft deflection angles, image transformation, block processing, temporal averaging, and post-visualization processing. Additionally, quantitative measurement of flow field velocity is achieved through tuft calibration. The study begins by comparing tufts of different materials, examining both fluorescent and deflection characteristics to determine optimal tuft parameters. The impact of tuft length on deflection characteristics is also investigated. Finally, the cotton tuft with a length of 7 mm and a diameter of about 0.1 mm was obtained as the best tuft sought in this experiment. Subsequently, at Reynolds number Re = 1.7 × 105, airfoil surface flow field visualization and quantitative analysis are conducted, exploring the relationship between airfoil surface flow structure and aerodynamics at different angles of attack. Finally, a comparative study is conducted between tuft velocity results and oil flow visualization experiments, as well as particle image velocimetry experiments, confirming the feasibility of the fluorescent tuft velocity measurement method.

Effect of concentration and wavelength of excitation on the photophysics of salicylate anion in acetonitrile and water

Sodium salicylate (NaSA) molecule exists as salicylate anion in acetonitrile (ACN) and water solvents, and exhibits large Stokes shifted fluorescence due to excited state intramolecular proton transfer (ESIPT), with decay times of ∼5 ns in ACN and ∼3.9 ns in water by 300 nm (absorption maxima) excitation. Previous studies report both ground and excited state enol-keto tautomerization in ACN, but only excited state tautomerization in water at 10−4 M. However, the current work explores the effect of concentration and excitation wavelengths on the photoinduced dynamics of ESIPT in the salicylate anion. On increasing the concentration of NaSA, no change in absorption spectra appears in both the solvents, and emission spectra of NaSA in water remain unaffected by changes in concentration or excitation wavelength, whereas, a slight red shift and decrease in FWHM appears in ACN. Time-domain fluorescence measurements show predominantly single-exponential decay throughout the emission profile by 300 nm excitation above the 10−5 M concentration in both the solvents, while by 375 nm excitation, multi-exponential fluorescence decay is observed at low concentrations, and as the concentration of NaSA increases, this decay behaviour tends to converge towards a single exponential decay. These results suggest that solute–solvent interactions stabilize the ground-state intermolecular hydrogen-bonded species at low concentrations, while higher concentrations weaken these interactions, leading to emission solely from the salicylate anion. Peak fit analysis of excitation spectra confirms enol-keto tautomerization in both the solvents, with the keto form being more stabilized in ACN. These findings underscore that in ACN, behaviour of NaSA is influenced by both concentration and excitation wavelength and contrary to previous reports, the keto form of the molecule is also present in water, though at a very low concentration and an increase in non-radiative transitions in water cause fluorescence quenching.

Development of red to blue emissive fluorescent materials for cyanide ion sensing and single-component inks in digital printing application

The work designed and described here is for the effective recognition of cyanide ion. A novel core (PC) with coumarin moiety as a signaling unit and pyridinium as an acceptor unit and these moieties linked by π-conjugation. In which CN− ion binds in the pyridine ring via nucleophilic addition and produces changes that are captured by different techniques to prove its sensing efficiency. The probe displays a visual color variation to pale orange from purple and also exhibits blue fluorescence from red fluorescence upon cyanide addition. The probe PC sensing ability towards CN− ion has been confirmed by various spectral measurement. In UV–visible technique, a blue shift was observed by the probe with CN− ion. In fluorescence measurements, a peak appeared at 704 nm, which corresponds to the probe encountering a peak enhancement with the added cyanide ion. The binding stoichiometric ratio among the probe and CN− ion is to be 1:1 in jobs method. The probes detection limit and binding constant were calculated to be 0.39 nM and 7.07x104 M. The probe is used as a single component ink and injected into a cartridge and printed on paper which is used as a tool to detect cyanide in water. Besides, the probe successfully senses cyanide ion in various water samples.

On the nature of initial solvation in bulk polar liquids: Gaussian or exponential?

Measurement of time evolution of fluorescence of a probe solute has been a quintessential technique to quantify how dipolar solvent molecules dynamically minimize the free energy of an electronically excited probe. During such solvation dynamics in bulk liquids, a substantial part of relaxation was shown to complete within sub-100 fs from time-gated fluorescence measurements, as also predicted by molecular dynamics simulation studies. However, equivalent quantification of solvation timescales by femtosecond pump-probe and broadband fluorescence measurements revealed an exponential nature of this initial relaxation having quite different timescales. Here, we set out to unveil the reason behind these puzzling contradictions. We introduce a method for estimating probe wavelength-dependent instrument response and demonstrate that the observation of the Gaussian vs exponential nature of initial relaxation is indeed dependent on the method of data analysis. These findings call for further experimental investigation and parallel development of theoretical models to elucidate the molecular-level mechanism accounting for different types of early time solvation.

Ultrasensitive strategy for OTA analysis by both recognition-activated multiple isothermal cyclic strand displacement amplification and DNAzyme-mediated cyclic signal reporting mechanism

We propose a recognition-driven ultrasensitive fluorescence sensing strategy for accurate analysis of ochratoxin A (OTA) by taking Mg2+-dependent DNAzyme based catalysis as the final signal output module. Recognition of target OTA destroys the simple double-strand recognition structure and contributes to the liberation of primer probe, which can trigger and activate the primer-mediated multiple isothermal cyclic strand displacement amplification (M-CSDA) process. The designed dual templates (T-P, T-A) can contribute to the generation of enormous nicked products (DNAzyme and primer), which will take effect in the triggering of M-CSDA. Meanwhile, one of the nicked products (DNAzyme) will also take part in the cyclically cleavage of designed hairpin probes (HP) with the presence of the cofactor Mg2+, inducing the recovery of fluorescent signal of quenched hairpin probes. Only the trace amount of target OTA could generate drastic fluorescent signal and ultrasensitive detection of OTA could be well achieved. Under optimized conditions, the detection limit of this method for OTA is 0.59 fg/mL. The characteristics of this method including the simplicity in dual signal amplification design, isothermal operation process and easy fluorescence measurement model exhibit the superior sensitivity and specificity for OTA detection. And this strategy can be further expanded for the design of ultrasensitive aptamer-sensing methods for series analytes.

Synthesis and functionalization of red-emissive carbon dots towards sensing of copper(II) and ATP in aqueous media

Red-emissive carbon dots, RCDs, were prepared in a bottom-up approach, from citric acid and urea, through a single solvothermal step, which were posteriorly surface-functionalized with di-(2-picolyl)amine (DPA) groups. The functionalized RCDs, henceforth named CD1, were analyzed through high-resolution transmission electron microscopy (HR-TEM), X-ray diffraction (XRD), dynamic light-scattering (DLS) and infrared spectroscopy (FTIR), and were found to be spherical, with diameter of circa 4 nm. UV-Vis and luminescence spectroscopic studies showed that CD1 display an absorption maximum at 550 nm, with emission maxima at around 640 nm. Fluorescence measurements indicated that Cu(II) induces the strongest changes in the emission of CD1 among eleven investigated metal ions, with a full quenching effect being observed. Further addition of an anionic species, adenosine 5-triphosphate (ATP), resulted in a recovery of the luminescence, which suggests that ATP is capable of sequestering metal ions from the surface of CD1. These results highlight the potential of simple and easy-to-make carbon nanomaterials as sensors for ions in aqueous media.

Dependence of photophysical properties on stereoisomers: The case of nonsymmetric 2-aryl(benz)imidazole-annulated derivatives

Nonsymmetric 2-aryl (benz)imidazole derivatives were previously synthesized via C-H/N-H annulation of imidazole derivatives from lapachol moieties with internal alkynes as an efficient tool to increase the π-system of lapimidazole derivatives. To investigate their photophysical properties, time-resolved and steady-state fluorescence measurements were carried out on dropcast film, powder, and diluted solution. Despite having the same number of single and double bonds, slight differences in their structures lead to distinct photophysical properties. The time-correlated single-photon counting measurements also demonstrated that the aspects of the temporal decay curves are dependent on the treatment of the samples during the measurements. The decay curves were relatively well fitted using an alternative method based on the exponentially modified Gaussian function, giving rise to a more complete set of fitting parameters. Finally, theoretical calculations corroborate that small structural changes in molecules can significantly alter their conformational dynamics, electronic spectra, and overall electronic behavior and stability.

Screening New Mungbean Varieties for Terminal Drought Tolerance

Rainfed mungbean crops in Queensland Australia frequently experience terminal drought (drought stress in the final stages of reproductive development), highlighting the importance of drought-tolerant varieties for sustainable mungbean production. Given there is limited information on the relative drought tolerance of current mungbean varieties in Australia, the study of genetic variations and mechanisms of drought tolerance in summer mungbean can provide a basis for developing drought-tolerant mungbean varieties. This study evaluated the physiological, biochemical, and phenological traits underpinning yield attributes associated with drought tolerance in selected mungbean varieties. Four new mungbean varieties (AVTMB#1 to 4) and the Australian commercial line (Jade-AU) were grown in tall (75 cm) polyvinyl chloride (PVC) lysimeters where drought stress was imposed at the early flowering stage (R1) and maintained until maturity. Drought stress significantly impacted all the varieties. Averaged across all the varieties, drought stress was associated with a reduction in stomatal conductance (gs) and photosynthetic rate (Asat) by 78% and 86%, respectively, compared to well-watered plants. Internal carbon dioxide concentration (Ci), the effective quantum yield of photosystem II (ΦPSII) and maximum light-use efficiency of light-acclimated photosystem II (PSII) centres (Fv’/Fm’) were also decreased, while excitation pressure (1-qP) increased with drought treatment. A positive correlation (r = 0.60) existed between seed yield and ΦPSII assessed at R1, while a weak correlation with Fv’/Fm’ (r = 0.24) was observed. Excitation pressure (1-qP) at the R1 stage was negatively correlated with seed yield (r = −0.66). Therefore, leaf fluorescence measures, viz., 1-qP and ΦPSII, were recommended for use in screening mungbean varieties for drought tolerance. The varieties, AVTMB#1 and AVTMB#4, respectively achieved 39 and 38% greater seed yields relative to the commercial variety, Jade-AU, under terminal drought conditions.