Chlorophyll Molecules Research Articles

Controlling Photosynthetic Excitons by Selective Pigment Photooxidation.

As a basis of photosynthesis, photoinduced oxidation of (bacterio)chlorophyll molecules in the special reaction center complexes has been a subject of extensive research. In contrast, the generally harmful photooxidation of antenna chromoproteins has received much less attention. Here, we have established the permanent structural changes in the LH2 antenna bacteriochlorophyll-protein complex from a sulfur photosynthetic purple bacterium Ectothiorhodospira haloalkaliphila taking place at physiological conditions upon intense optical irradiation. To this end, a crystal structure of the LH2 complex from E. haloalkaliphila was first resolved by X-ray diffraction to 3.7 Å, verifying a great similarity with the earlier structure from Phaesporillum molischianum. Analysis of the various steady-state and picosecond time-resolved optical spectroscopy data and related model simulations then confirmed that the major spectral effects observed-bleaching and blue-shifting of the B850 exciton band and correlated emergence of a higher-energy C700 exciton band-are associated with photooxidation of increasing numbers of B850 bacteriochlorophylls into 3-acetyl-chlorophylls, with no noticeable damage to the pigment-binding protein scaffold. A prospective noninvasive method for an in situ optical control of excitons by selective photooxidation of pigment chromophores was thus revealed and demonstrated in a structurally well-defined native system.

Chlorophyll degradation by tetracycline and cadmium in spinach (Spinacia oleracea L.) leaves

Degradation of chlorophyll was studied in leaves of spinach grown in soil containing cadmium and tetracycline, based on spectroscopic measurements and biochemical analyses of plant extracts. It was shown that plant exposure to the highest levels of tetracycline and cadmium resulted in 64% and 68%, respectively, reduction in chlorophyll content. The chlorophyll degradation rate constants were determined, and they were found to increase with increasing doses of tetracycline and cadmium. The rate constant of chlorophyll degradation by tetracycline ranged from k = 960 M−1 day−1 to k = 2180 M−1 day−1, and the rate constant of chlorophyll degradation by cadmium ranged from k = 1130 M−1 day−1 to k = 2580 M−1 day−1, depending on dose. Plant stress responses to tetracycline are much stronger than to cadmium, as is visible from the activity of guaiacol peroxidase and catalase. However, phytotoxicity of cadmium, measured by the rate of chlorophyll degradation and enzyme activities, is much higher compared to tetracycline. The spectroscopic measurements were taken 10 days after tetracycline and cadmium were added to the reagent grade chlorophyll which was used at the concentration of chlorophyll in plants. Changes in absorption and fluorescence spectra are likely to result from removal of magnesium from the chlorophyll molecule, and thus they indicate the formation of pheophytin. Cadmium, on the other hand, is probably bound into the chlorophyll molecule, substituting its magnesium.

Time-resolved fluorescence measurements on leaves: principles and recent developments

Photosynthesis starts when a pigment in the photosynthetic antennae absorbs a photon. The electronic excitation energy is then transferred through the network of light-harvesting pigments to special chlorophyll (Chl) molecules in the reaction centres, where electron transfer is initiated. Energy transfer and primary electron transfer processes take place on timescales ranging from femtoseconds to nanoseconds, and can be monitored in real time via time-resolved fluorescence spectroscopy. This method is widely used for measurements on unicellular photosynthetic organisms, isolated photosynthetic membranes, and individual complexes. Measurements on intact leaves remain a challenge due to their high structural heterogeneity, high scattering, and high optical density, which can lead to optical artefacts. However, detailed information on the dynamics of these early steps, and the underlying structure–function relationships, is highly informative and urgently required in order to get deeper insights into the physiological regulation mechanisms of primary photosynthesis. Here, we describe a current methodology of time-resolved fluorescence measurements on intact leaves in the picosecond to nanosecond time range. Principles of fluorescence measurements on intact leaves, possible sources of alterations of fluorescence kinetics and the ways to overcome them are addressed. We also describe how our understanding of the organisation and function of photosynthetic proteins and energy flow dynamics in intact leaves can be enriched through the application of time-resolved fluorescence spectroscopy on leaves. For that, an example of a measurement on Zea mays leaves is presented.

Photophysical and photocatalytic properties of corrophyll and chlorophyll

Tetrapyrrolic macrocycles are well known photosensitizers, absorbing light in the ultraviolet and visible region. The quantum efficiency of these naturally occurring compounds in optical processes, and the possibility of altering their behavior by modifying its constituent features, make them promising candidates for applications as Dye Sensitized Solar Cells (DSSC) and photocatalytic reactions. The time-dependent density-functional theory (TD-DFT) were used to study the optical and redox properties of chlorophyll and corrin-related molecules (corrophyll). The influence of the substituents and metallic atoms in their properties have been investigated. Our results show lower reduction potentials for corrophyll molecules compared with chlorophyll. The optical absorbance spectra of corrophyll without a metallic atom at their central rings shows a significant blue-shift, as compared to their chlorophyll counterparts. The presence of Co(I) ion species at the corrophyll core leads to oxidation potentials below than that of water, which puts corrophyll ahead of traditional chlorophyll pigments as photocatalysts. We show that the substituents and the ions coordinated to the macrocycles play an important role in this phenomena. These findings show the great potential of tuning the spectroscopic and reactive properties of tetrapyrrole macrocycles for applications in photocatalysis and optoelectronic devices, while keeping their essential structural features intact.

Chlorophyll-Carotenoid Excitation Energy Transfer in High-Light-Exposed Thylakoid Membranes Investigated by Snapshot Transient Absorption Spectroscopy.

Nonphotochemical quenching (NPQ) provides an essential photoprotection in plants, assuring safe dissipation of excess energy as heat under high light. Although excitation energy transfer (EET) between chlorophyll (Chl) and carotenoid (Car) molecules plays an important role in NPQ, detailed information on the EET quenching mechanism under in vivo conditions, including the triggering mechanism and activation dynamics, is very limited. Here, we observed EET between the Chl Q y state and the Car S1 state in high-light-exposed spinach thylakoid membranes. The kinetic and spectral analyses using transient absorption (TA) spectroscopy revealed that the Car S1 excited state absorption (ESA) signal after Chl excitation has a maximum absorption peak around 540 nm and a lifetime of ∼8 ps. Snapshot TA spectroscopy at multiple time delays allowed us to track the Car S1 ESA signal as the thylakoid membranes were exposed to various light conditions. The obtained snapshots indicate that maximum Car S1 ESA signal quickly rose and slightly dropped during the initial high-light exposure (<3 min) and then gradually increased with a time constant of ∼5 min after prolonged light exposure. This suggests the involvement of both rapidly activated and slowly activated mechanisms for EET quenching. 1,4-Dithiothreitol (DTT) and 3,3'-dithiobis(sulfosuccinimidyl propionate) (DTSSP) chemical treatments further support that the Car S1 ESA signal (or the EET quenching mechanism) is primarily dependent on the accumulation of zeaxanthin and partially dependent on the reorganization of membrane proteins, perhaps due to the pH-sensing protein photosystem II subunit S.

Ultrafast Energy Transfer Involving the Red Chlorophylls of Cyanobacterial Photosystem I Probed through Two-Dimensional Electronic Spectroscopy.

Photosystem I (PSI) is a naturally occurring light-harvesting complex that drives oxygenic photosynthesis through a series of photoinitiated transmembrane electron transfer reactions that occur with a high quantum efficiency. Understanding the mechanism by which this process occurs is fundamental to understanding the near-unity quantum efficiency of PSI and in turn could lead to further insight into PSI-based technologies for solar energy conversion. In this article, we have applied two-dimensional electronic spectroscopy to PSI complexes isolated from two different cyanobacterial strains to gain further insight into the ultrafast energy transfer in PSI. The PSI complexes studied differ in the number and absorption of the red chlorophylls, chlorophylls that lie to lower energies than the reaction center. By applying a global analysis to the 2D electronic spectra of the PSI complexes we extract 2D decay associated spectra (2D-DAS). Through analysis of the 2D-DAS we observe a 50 fs relaxation among the bulk antenna chlorophylls in addition to two pathways of energy equilibration involving the red chlorophylls: a fast 200 fs equilibration followed by a 2-4 ps equilibration. As demonstrated with a model system, the λ1, λ3 coordinates of the cross-peaks in the 2D-DAS spectra indicate that the two equilibration pathways involve different chlorophyll molecules.

Intramolecular Vibrations Complement the Robustness of Primary Charge Separation in a Dimer Model of the Photosystem II Reaction Center.

The energy conversion of oxygenic photosynthesis is triggered by primary charge separation in proteins at the photosystem II reaction center. Here, we investigate the impacts of the protein environment and intramolecular vibrations on primary charge separation at the photosystem II reaction center. This is accomplished by combining the quantum dynamic theories of condensed phase electron transfer with quantum chemical calculations to evaluate the vibrational Huang-Rhys factors of chlorophyll and pheophytin molecules. We report that individual vibrational modes play a minor role in promoting charge separation, contrary to the discussion in recent publications. Nevertheless, these small contributions accumulate to considerably influence the charge separation rate, resulting in subpicosecond charge separation almost independent of the driving force and temperature. We suggest that the intramolecular vibrations complement the robustness of the charge separation in the photosystem II reaction center against the inherently large static disorder of the involved electronic energies.

Optical, spectral and photovoltaic characterization of natural dyes extracted from leaves of Peltophorum pterocarpum and Acalypha amentacea used as sensitizers for ZnO based dye sensitized solar cells

The ZnO nanorods were synthesized using hydrothermal method. The morphologies, crystalline nature and optical properties of the synthesized ZnO nanorods were characterized by X-ray diffractometer, High-resolution scanning electron microscopy [HRSEM], and UV–Vis absorption spectroscopic studies. A thin layer of ZnO nanorods were spread on transparent conducting FTO coated glass plate using Doctor Blade method. Natural dyes extracted from Peltophorum pterocarpum and Acalypha amentacea leaves with solvents such as water and ethanol were used as sensitizers to fabricate dye sensitized solar cells (DSSCs). The extracted dyes were characterized by Fourier transform infrared (FTIR) and UV–Visible absorption spectroscopy. The FTIR confirms the presence of anthocyanin, carotene and chlorophyll molecules in the dye extracts. The influence on the different parameters such as open circuit voltage (VOC) short circuit current (JSC), fill factor (FF), and power conversion efficiency (η) of the dye sensitized solar cells (DSSCs) were analyzed. The ZnO nanorods provide high specific surface area for dye adsorption and the pathway is efficient for electron transportation.

A gene co-expression network model identifies yield-related vicinity networks in Jatropha curcas shoot system

The plant shoot system consists of reproductive organs such as inflorescences, buds and fruits, and the vegetative leaves and stems. In this study, the reproductive part of the Jatropha curcas shoot system, which includes the aerial shoots, shoots bearing the inflorescence and inflorescence were investigated in regard to gene-to-gene interactions underpinning yield-related biological processes. An RNA-seq based sequencing of shoot tissues performed on an Illumina HiSeq. 2500 platform generated 18 transcriptomes. Using the reference genome-based mapping approach, a total of 64 361 genes was identified in all samples and the data was annotated against the non-redundant database by the BLAST2GO Pro. Suite. After removing the outlier genes and samples, a total of 12 734 genes across 17 samples were subjected to gene co-expression network construction using petal, an R library. A gene co-expression network model built with scale-free and small-world properties extracted four vicinity networks (VNs) with putative involvement in yield-related biological processes as follow; heat stress tolerance, floral and shoot meristem differentiation, biosynthesis of chlorophyll molecules and laticifers, cell wall metabolism and epigenetic regulations. Our VNs revealed putative key players that could be adapted in breeding strategies for J. curcas shoot system improvements.

Photochemistry beyond the red limit in chlorophyll f-containing photosystems.

Photosystems I and II convert solar energy into the chemical energy that powers life. Chlorophyll a photochemistry, using red light (680 to 700 nm), is near universal and is considered to define the energy "red limit" of oxygenic photosynthesis. We present biophysical studies on the photosystems from a cyanobacterium grown in far-red light (750 nm). The few long-wavelength chlorophylls present are well resolved from each other and from the majority pigment, chlorophyll a. Charge separation in photosystem I and II uses chlorophyll f at 745 nm and chlorophyll f (or d) at 727 nm, respectively. Each photosystem has a few even longer-wavelength chlorophylls f that collect light and pass excitation energy uphill to the photochemically active pigments. These photosystems function beyond the red limit using far-red pigments in only a few key positions.

Leaf wound induced ultraweak photon emission is suppressed under anoxic stress: Observations of Spathiphyllum under aerobic and anaerobic conditions using novel in vivo methodology.

Plants have evolved a variety of means to energetically sense and respond to abiotic and biotic environmental stress. Two typical photochemical signaling responses involve the emission of volatile organic compounds and light. The emission of certain leaf wound volatiles and light are mutually dependent upon oxygen which is subsequently required for the wound-induced lipoxygenase reactions that trigger the formation of fatty acids and hydroperoxides; ultimately leading to photon emission by chlorophyll molecules. A low noise photomultiplier with sensitivity in the visible spectrum (300–720 nm) is used to continuously measure long duration ultraweak photon emission of dark-adapting whole Spathiphyllum leaves (in vivo). Leaves were mechanically wounded after two hours of dark adaptation in aerobic and anaerobic conditions. It was found that (1) nitrogen incubation did not affect the pre-wound basal photocounts; (2) wound induced leaf biophoton emission was significantly suppressed when under anoxic stress; and (3) the aerobic wound induced emission spectra observed was > 650 nm, implicating chlorophyll as the likely emitter. Limitations of the PMT photocathode’s radiant sensitivity, however, prevented accurate analysis from 700–720 nm. Further examination of leaf wounding profile photon counts revealed that the pre-wounding basal state (aerobic and anoxic), the anoxic wounding state, and the post-wounding aerobic state statistics all approximate a Poisson distribution. It is additionally observed that aerobic wounding induces two distinct exponential decay events. These observations contribute to the body of plant wound-induced luminescence research and provide a novel methodology to measure this phenomenon in vivo.

Lower-energy photons do the work, too

Photosynthesis Plants and cyanobacteria use chlorophyll-rich photosystem complexes to convert light energy into chemical energy. Some organisms have developed adaptations to take advantage of longer-wavelength photons. Nurnberg et al. studied photosystem complexes from cyanobacteria grown in the presence of far-red light. The authors identified the primary donor chlorophyll as one of a few chlorophyll molecules in the far-red light–adapted enzymes that were chemically altered to shift their absorption spectrum. Kinetic measurements demonstrated that far-red light is capable of directly driving water oxidation, despite having less energy than the red light used by most photosynthetic organisms. Science , this issue p. [1210][1] [1]: /lookup/doi/10.1126/science.aar8313

Development of fluorescence quenching in Chlamydomonas reinhardtii upon prolonged illumination at 77\xa0K

Low-temperature fluorescence measurements are frequently used in photosynthesis research to assess photosynthetic processes. Upon illumination of photosystem II (PSII) frozen to 77 K, fluorescence quenching is observed. In this work, we studied the light-induced quenching in intact cells of Chlamydomonas reinhardtii at 77 K using time-resolved fluorescence spectroscopy with a streak camera setup. In agreement with previous studies, global analysis of the data shows that prolonged illumination of the sample affects the nanosecond decay component of the PSII emission. Using target analysis, we resolved the quenching on the PSII-684 compartment which describes bulk chlorophyll molecules of the PSII core antenna. Further, we quantified the quenching rate constant and observed that as the illumination proceeds the accumulation of the quencher leads to a speed up of the fluorescence decay of the PSII-684 compartment as the decay rate constant increases from about 3 to 4 ns− 1. The quenching on PSII-684 leads to indirect quenching of the compartments PSII-690 and PSII-695 which represent the red chlorophyll of the PSII core. These results explain past and current observations of light-induced quenching in 77 K steady-state and time-resolved fluorescence spectra.

Singlet oxygen-triggered chloroplast-to-nucleus retrograde signalling pathways: An emerging perspective.

Singlet oxygen (1 O2 ) is a prime cause of photo-damage of the photosynthetic apparatus. The chlorophyll molecules in the photosystem II reaction center and in the light-harvesting antenna complex are major sources of 1 O2 generation. It has been thought that the generation of 1 O2 mainly takes place in the appressed regions of the thylakoid membranes, namely, the grana core, where most of the active photosystem II complexes are localized. Apart from being a toxic molecule, new evidence suggests that 1 O2 significantly contributes to chloroplast-to-nucleus retrograde signalling that primes acclimation and cell death responses. Interestingly, recent studies reveal that chloroplasts operate two distinct 1 O2 -triggered retrograde signalling pathways in which β-carotene and a nuclear-encoded chloroplast protein EXECUTER1 play essential roles as signalling mediators. The coexistence of these mediators raises several questions: their crosstalk, source(s) of 1 O2 , downstream signalling components, and the perception and reaction mechanism of these mediators towards 1 O2 . In this review, we mainly discuss the molecular genetic basis of the mode of action of these two putative 1 O2 sensors and their corresponding retrograde signalling pathways. In addition, we also propose the possible existence of an alternative source of 1 O2 , which is spatially and functionally separated from the grana core.

Structure of the maize photosystem I supercomplex with light-harvesting complexes I and II.

Plants regulate photosynthetic light harvesting to maintain balanced energy flux into photosystems I and II (PSI and PSII). Under light conditions favoring PSII excitation, the PSII antenna, light-harvesting complex II (LHCII), is phosphorylated and forms a supercomplex with PSI core and the PSI antenna, light-harvesting complex I (LHCI). Both LHCI and LHCII then transfer excitation energy to the PSI core. We report the structure of maize PSI-LHCI-LHCII solved by cryo-electron microscopy, revealing the recognition site between LHCII and PSI. The PSI subunits PsaN and PsaO are observed at the PSI-LHCI interface and the PSI-LHCII interface, respectively. Each subunit relays excitation to PSI core through a pair of chlorophyll molecules, thus revealing previously unseen paths for energy transfer between the antennas and the PSI core.

Mass Spectrometry of Non-allomerized Chlorophylls a and b Derivatives from Plants

Background: Non-allomerized chlorophylls a and b are part of a great diversity of environments: senescent and ripened vegetal tissues, processed foods, sediments, fecal pellets, etc. Consequently, the articles characterizing their mass spectrometry behavior belong to a variety of disciplines. Objective: To compile in a single review all the information published related to mass spectrometry of 26 chlorophyll derivatives, including chlorophyllides, pheophorbides, pheophytins, and hydroxy-, lactone-, and pyro-chlorophyll derivatives. Method: A detailed and extensive bibliographic search through the Web of ScienceTM followed by a comprehensive and critic discussion. Results: This review gathers all the information concerning mass spectroscopic characteristics published for each non-allomerized chlorophyll of a and b series. This means that for each one of the 26 chlorophyll derivatives, it has been compiled the complete information published of mass (or exact mass), mass spectrometry technique and equipment (ionization method and mass analyzer), product ions, structural assignments and proposed mechanisms (if available), detailing the references. Conclusion: The evaluation of referenced data in literature has allowed to obtain common characteristics in the mass spectrometry behavior between the different chlorophyll derivatives. In addition, specific product ions have been reported, revealing specific structural modifications in the chlorophyll molecule, facilitating the structural characterization. This review embraces the application of predictive software tools, which improve accuracy of predictions and make straightforward the structural assignments. Keywords: Chlorophyllide, hydroxy-chlorophyll, lactone-chlorophyll, mass spectrometry, pheophorbide, pheophytin, pyrochlorophyll.

A MODEL TO SIMULATE THE RADIATIVE TRANSFER OF FLUORESCENCE IN A LEAF

Abstract. Light is reflected, transmitted and absorbed by green leaves. Chlorophyll fluorescence (ChlF) is the signal emitted by chlorophyll molecules in the leaf after the absorption of light. ChlF can be used as a direct probe of the functional status of photosynthetic machinery because of its close relationship with photosynthesis. The scattering, absorbing, and emitting properties of leaves are spectrally dependent, which can be simulated by modeling leaf-level fluorescence. In this paper, we proposed a Monte-Carlo (MC) model to simulate the radiative transfer of photons in the leaf. Results show that typical leaf fluorescence spectra can be properly simulated, with two peaks centered at around 685 nm in the red and 740 nm in the far-red regions. By analysing the sensitivity of the input parameters, we found the MC model can well simulate their influence on the emitted fluorescence. Meanwhile we compared results simulated by MC model with those by the Fluspect model. Generally they agree well in the far-red region but deviate in the red region.

Cell size, photosynthesis and the package effect: an artificial selection approach.

Cell size correlates with most traits among phytoplankton species. Theory predicts that larger cells should show poorer photosynthetic performance, perhaps due to reduced intracellular self-shading (i.e. package effect). Yet current theory relies heavily on interspecific correlational approaches and causal relationships between size and photosynthetic machinery have remained untested. As a more direct test, we applied 250 generations of artificial selection (c. 20months) to evolve the green microalga Dunaliella teriolecta (Chlorophyta) toward different mean cell sizes, while monitoring all major photosynthetic parameters. Evolving larger sizes (>1500% difference in volume) resulted in reduced oxygen production per chlorophyll molecule - as predicted by the package effect. However, large-evolved cells showed substantially higher rates of oxygen production - a finding unanticipated by current theory. In addition, volume-specific photosynthetic pigments increased with size (Chla+b), while photo-protectant pigments decreased (β-carotene). Finally, larger cells displayed higher growth performances and Fv /Fm , steeper slopes of rapid light curves (α) and smaller light-harvesting antennae (σPSII ) with higher connectivity (ρ). Overall, evolving a common ancestor into different sizes showed that the photosynthetic characteristics of a species coevolves with cell volume. Moreover, our experiment revealed a trade-off between chlorophyll-specific (decreasing with size) and volume-specific (increasing with size) oxygen production in a cell.

The Ribosome-Bound Protein Pam68 Promotes Insertion of Chlorophyll into the CP47 Subunit of Photosystem II.

Photosystem II (PSII) is a large enzyme complex embedded in the thylakoid membrane of oxygenic phototrophs. The biogenesis of PSII requires the assembly of more than 30 subunits, with the assistance of a number of auxiliary proteins. In plants and cyanobacteria, the photosynthesis-affected mutant 68 (Pam68) is important for PSII assembly. However, its mechanisms of action remain unknown. Using a Synechocystis PCC 6803 strain expressing Flag-tagged Pam68, we purified a large protein complex containing ribosomes, SecY translocase, and the chlorophyll-binding PSII inner antenna CP47. Using 2D gel electrophoresis, we identified a pigmented Pam68-CP47 subcomplex and found Pam68 bound to ribosomes. Our results show that Pam68 binds to ribosomes even in the absence of CP47 translation. Furthermore, Pam68 associates with CP47 at an early phase of its biogenesis and promotes the synthesis of this chlorophyll-binding polypeptide until the attachment of the small PSII subunit PsbH. Deletion of both Pam68 and PsbH nearly abolishes the synthesis of CP47, which can be restored by enhancing chlorophyll biosynthesis. These results strongly suggest that ribosome-bound Pam68 stabilizes membrane segments of CP47 and facilitates the insertion of chlorophyll molecules into the translated CP47 polypeptide chain.

Regulating Photonic Properties of Lamellar Chloroplast and the Environmental Adaption

Iridescent structures in photosynthetic organisms has recently been identified to possess enhanced light harvesting capability. It was proposed that interference nature of periodic photonic nanostructure results in this enhancement, nonetheless, it will inevitably accompany with light capture efficiency reduction in the photonic band gap spectrum range. Enhancement or reduction of light capture efficiency depends on the coupling condition between the electric field spatial variation and the lamellar structures. This work will provide insight about how photonic band gap wavelength of the lamellae can be regulated and the possibility in correlation between this regulation and different strategies from nature for environmental adaptation. Most of the plants with iridescent chloroplasts dwell in understorey environment, low light coherence condition might affect the predicted efficiency enhancement, and some results will be presented. These results will offer new perception about the formation of lamellar structure become a better tactic than accumulate more chlorophyll (or other light absorbing materials) molecules together for obtaining better light harvesting efficiency.