Nejvíce citovaný článek - PubMed ID 10354490
Chlorophyll a fluorescence induction1
Quantitative measurement of light intensity is a key step in ensuring the reliability and the reproducibility of scientific results in many fields of physics, biology, and chemistry. The protocols presented so far use various photoactive properties of manufactured materials. Here, leaves are introduced as an easily accessible green material to calibrate light intensity. The measurement protocol consists in monitoring the chlorophyll fluorescence of a leaf while it is exposed to a jump of constant light. The inverse of the characteristic time of the initial chlorophyll fluorescence rise is shown to be proportional to the light intensity received by the leaf over a wide range of wavelengths and intensities. Moreover, the proportionality factor is stable across a wide collection of plant species, which makes the measurement protocol accessible to users without prior calibration. This favorable feature is finally harnessed to calibrate a source of white light from exploiting simple leaves collected from a garden.
- Klíčová slova
- actinometry, fluorescence, green materials, irradiance, light intensity, photoactive materials,
- Publikační typ
- časopisecké články MeSH
Plants growing in nature often experience fluctuating irradiance. However, in the laboratory, the dynamics of photosynthesis are usually explored by instantaneously exposing dark-adapted plants to constant light and examining the dark-to-light transition, which is a poor approximation of natural phenomena. With the aim creating a better approximation, we exposed leaves of pea (Pisum sativum) to oscillating light and measured changes in the functioning of PSI and PSII, and of the proton motive force at the thylakoid membrane. We found that the dynamics depended on the oscillation period, revealing information about the underlying regulatory networks. As demonstrated for a selected oscillation period of 60 s, the regulation tries to keep the reaction centers of PSI and PSII open. We present an evaluation of the data obtained, and discuss the involvement of particular processes in the regulation of photosynthesis. The forced oscillations provided an information-rich fingerprint of complex regulatory networks. We expect future progress in understanding these networks from experiments involving chemical interventions and plant mutants, and by using mathematical modeling and systems identification and control tools.
- Klíčová slova
- Pisum sativum, Fluctuating light, forced oscillations, pea, photosynthesis, photosystem I and II, proton motive force, regulation,
- MeSH
- fotosyntéza fyziologie MeSH
- fotosystém I (proteinový komplex) metabolismus MeSH
- fotosystém II (proteinový komplex) * metabolismus MeSH
- hrách setý * metabolismus MeSH
- listy rostlin metabolismus MeSH
- rostliny metabolismus MeSH
- světlo MeSH
- transport elektronů fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- fotosystém I (proteinový komplex) MeSH
- fotosystém II (proteinový komplex) * MeSH
Oxygenic photosynthesis takes place in thylakoid membranes (TM) of cyanobacteria, algae, and higher plants. It begins with light absorption by pigments in large (modular) assemblies of pigment-binding proteins, which then transfer excitation energy to the photosynthetic reaction centers of photosystem (PS) I and PSII. In green algae and plants, these light-harvesting protein complexes contain chlorophylls (Chls) and carotenoids (Cars). However, cyanobacteria, red algae, and glaucophytes contain, in addition, phycobiliproteins in phycobilisomes that are attached to the stromal surface of TM, and transfer excitation energy to the reaction centers via the Chl a molecules in the inner antennas of PSI and PSII. The color and the intensity of the light to which these photosynthetic organisms are exposed in their environment have a great influence on the composition and the structure of the light-harvesting complexes (the antenna) as well as the rest of the photosynthetic apparatus, thus affecting the photosynthetic process and even the entire organism. We present here a perspective on 'Light Quality and Oxygenic Photosynthesis', in memory of George Christos Papageorgiou (9 May 1933-21 November 2020; see notes a and b). Our review includes (1) the influence of the solar spectrum on the antenna composition, and the special significance of Chl a; (2) the effects of light quality on photosynthesis, measured using Chl a fluorescence; and (3) the importance of light quality, intensity, and its duration for the optimal growth of photosynthetic organisms.
The largest stable photosystem II (PSII) supercomplex in land plants (C2S2M2) consists of a core complex dimer (C2), two strongly (S2) and two moderately (M2) bound light-harvesting protein (LHCB) trimers attached to C2 via monomeric antenna proteins LHCB4-6. Recently, we have shown that LHCB3 and LHCB6, presumably essential for land plants, are missing in Norway spruce (Picea abies), which results in a unique structure of its C2S2M2 supercomplex. Here, we performed structure-function characterization of PSII supercomplexes in Arabidopsis (Arabidopsis thaliana) mutants lhcb3, lhcb6, and lhcb3 lhcb6 to examine the possibility of the formation of the "spruce-type" PSII supercomplex in angiosperms. Unlike in spruce, in Arabidopsis both LHCB3 and LHCB6 are necessary for stable binding of the M trimer to PSII core. The "spruce-type" PSII supercomplex was observed with low abundance only in the lhcb3 plants and its formation did not require the presence of LHCB4.3, the only LHCB4-type protein in spruce. Electron microscopy analysis of grana membranes revealed that the majority of PSII in lhcb6 and namely in lhcb3 lhcb6 mutants were arranged into C2S2 semi-crystalline arrays, some of which appeared to structurally restrict plastoquinone diffusion. Mutants without LHCB6 were characterized by fast induction of non-photochemical quenching and, on the contrary to the previous lhcb6 study, by only transient slowdown of electron transport between PSII and PSI. We hypothesize that these functional changes, associated with the arrangement of PSII into C2S2 arrays in thylakoids, may be important for the photoprotection of both PSI and PSII upon abrupt high-light exposure.
- MeSH
- Arabidopsis genetika metabolismus MeSH
- fotosystém II (proteinový komplex) genetika metabolismus MeSH
- proteiny huseníčku genetika metabolismus MeSH
- proteiny vázající chlorofyl genetika metabolismus MeSH
- smrk metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- fotosystém II (proteinový komplex) MeSH
- Lhcb6 protein, Arabidopsis MeSH Prohlížeč
- proteiny huseníčku MeSH
- proteiny vázající chlorofyl MeSH
Foundations of photosynthesis research have been established mainly by studying the response of plants to changing light, typically to sudden exposure to a constant light intensity after dark acclimation or light flashes. This approach remains valid and powerful, but can be limited by requiring dark acclimation before time-domain measurements and often assumes that rate constants determining the photosynthetic response do not change between dark and light acclimation. We show that these limits can be overcome by measuring plant responses to sinusoidally modulated light of varying frequency. By its nature, such frequency-domain characterization is performed in light-acclimated plants with no need for prior dark acclimation. Amplitudes, phase shifts, and upper harmonic modulation extracted from the data for a wide range of frequencies can target different kinetic domains and regulatory feedbacks. The occurrence of upper harmonic modulation reflects nonlinear phenomena, including photosynthetic regulation. To support these claims, we measured chlorophyll fluorescence emission of the green alga Chlorella sorokiniana in light that was sinusoidally modulated in the frequency range 1000-0.001 Hz. Based on these experimental data and numerical as well as analytical mathematical models, we propose that frequency-domain measurements can become a versatile tool in plant sensing.
In Part I, by using 31P-NMR spectroscopy, we have shown that isolated granum and stroma thylakoid membranes (TMs), in addition to the bilayer, display two isotropic phases and an inverted hexagonal (HII) phase; saturation transfer experiments and selective effects of lipase and thermal treatments have shown that these phases arise from distinct, yet interconnectable structural entities. To obtain information on the functional roles and origin of the different lipid phases, here we performed spectroscopic measurements and inspected the ultrastructure of these TM fragments. Circular dichroism, 77 K fluorescence emission spectroscopy, and variable chlorophyll-a fluorescence measurements revealed only minor lipase- or thermally induced changes in the photosynthetic machinery. Electrochromic absorbance transients showed that the TM fragments were re-sealed, and the vesicles largely retained their impermeabilities after lipase treatments-in line with the low susceptibility of the bilayer against the same treatment, as reflected by our 31P-NMR spectroscopy. Signatures of HII-phase could not be discerned with small-angle X-ray scattering-but traces of HII structures, without long-range order, were found by freeze-fracture electron microscopy (FF-EM) and cryo-electron tomography (CET). EM and CET images also revealed the presence of small vesicles and fusion of membrane particles, which might account for one of the isotropic phases. Interaction of VDE (violaxanthin de-epoxidase, detected by Western blot technique in both membrane fragments) with TM lipids might account for the other isotropic phase. In general, non-bilayer lipids are proposed to play role in the self-assembly of the highly organized yet dynamic TM network in chloroplasts.
- Klíčová slova
- SAXS, bilayer, chlorophyll fluorescence, cryo-electron-tomography, electron microscopy, membrane energization, membrane networks, non-bilayer lipid phases, violaxanthin de-epoxidase,
- MeSH
- cirkulární dichroismus metody MeSH
- elektronová mikroskopie metody MeSH
- fotosyntéza genetika MeSH
- lipidy genetika MeSH
- magnetická rezonanční spektroskopie metody MeSH
- tylakoidy genetika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- lipidy MeSH
BACKGROUND: With limited agricultural land and increasing human population, it is essential to enhance overall photosynthesis and thus productivity. Oxygenic photosynthesis begins with light absorption, followed by excitation energy transfer to the reaction centres, primary photochemistry, electron and proton transport, NADPH and ATP synthesis, and then CO2 fixation (Calvin-Benson cycle, as well as Hatch-Slack cycle). Here we cover some of the discoveries related to this process, such as the existence of two light reactions and two photosystems connected by an electron transport 'chain' (the Z-scheme), chemiosmotic hypothesis for ATP synthesis, water oxidation clock for oxygen evolution, steps for carbon fixation, and finally the diverse mechanisms of regulatory processes, such as 'state transitions' and 'non-photochemical quenching' of the excited state of chlorophyll a. SCOPE: In this review, we emphasize that mathematical modelling is a highly valuable tool in understanding and making predictions regarding photosynthesis. Different mathematical models have been used to examine current theories on diverse photosynthetic processes; these have been validated through simulation(s) of available experimental data, such as chlorophyll a fluorescence induction, measured with fluorometers using continuous (or modulated) exciting light, and absorbance changes at 820 nm (ΔA820) related to redox changes in P700, the reaction centre of photosystem I. CONCLUSIONS: We highlight here the important role of modelling in deciphering and untangling complex photosynthesis processes taking place simultaneously, as well as in predicting possible ways to obtain higher biomass and productivity in plants, algae and cyanobacteria.
- Klíčová slova
- Calvin–Benson cycle, chlorophyll a fluorescence induction, discoveries in photosynthesis, modelling, non-photochemical quenching (of the excited state of chlorophyll a), photosynthetic electron transport, state transitions,
- MeSH
- biomasa MeSH
- chlorofyl a * MeSH
- chlorofyl MeSH
- fotosyntéza * MeSH
- fotosystém II (proteinový komplex) MeSH
- kyslík MeSH
- lidé MeSH
- světlo MeSH
- transport elektronů MeSH
- voda MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- chlorofyl a * MeSH
- chlorofyl MeSH
- fotosystém II (proteinový komplex) MeSH
- kyslík MeSH
- voda MeSH
Miscanthus x giganteus (Mxg) is a promising second-generation biofuel crop with high production of energetic biomass. Our aim was to determine the level of plant stress of Mxg grown in poor quality soils using non-invasive physiological parameters and to test whether the stress could be reduced by application of plant growth regulators (PGRs). Plant fitness was quantified by measuring of leaf fluorescence using 24 indexes to select the most suitable fluorescence indicators for quantification of this type of abiotic stress. Simultaneously, visible stress signs were observed on stems and leaves and differences in variants were revealed also by microscopy of leaf sections. Leaf fluorescence analysis, visual observation and changes of leaf anatomy revealed significant stress in all studied subjects compared to those cultivated in good quality soil. Besides commonly used Fv/Fm (potential photosynthetic efficiency) and P.I. (performance index), which showed very low sensitivity, we suggest other fluorescence parameters (like dissipation, DIo/RC) for revealing finer differences. We can conclude that measurement of leaf fluorescence is a suitable method for revealing stress affecting Mxg in poor soils. However, none of investigated parameters proved significant positive effect of PGRs on stress reduction. Therefore, direct improvement of soil quality by fertilization should be considered for stress reduction and improving the biomass quality in this type of soils.
- Klíčová slova
- Miscanthus x giganteus, leaf fluorescence, nutritionally poor post-military soil, plant physiology, plant stress,
- Publikační typ
- časopisecké články MeSH
Short-term heat exposure in tropical regions can generate severe stress in the photosynthetic activity of soil crust cyanobacteria. We investigated the responses of two filamentous cyanobacteria, Scytonema tolypothrichoides and Tolypothrix bouteillei, to 1hr exposure at 35, 45, and 55 °C using variable chlorophyll fluorescence. Protocols for maximum quantum yield (FV/FM) and dark recovery of chlorophyll a fluorescence (OJIP) transient were applied. Heat exposure caused damage to the donor side of PSII, indicated by a decrease in FV/FM and a rapid increase in F0. After heat stress, photochemical energy utilization (φPo, φETo, and φRE1o) declined and energy dissipation (φDIo) increased. At 45 °C, the photosynthetic apparatus was reversibly damaged, since full recovery was observed after 7 days of relaxation. S. tolypothrichoides was more resistant to heat stress than T. bouteillei, confirming better adaptation to higher temperatures as observed in growth experiments.
- Klíčová slova
- Cyanobacteria, Heat stress, OJIP transient, Photosynthesis,
- MeSH
- bakteriální proteiny metabolismus MeSH
- časové faktory MeSH
- chlorofyl a metabolismus MeSH
- fluorescence MeSH
- fotosyntetická reakční centra (proteinové komplexy) metabolismus MeSH
- fotosyntéza fyziologie MeSH
- kvantová teorie MeSH
- pravděpodobnost MeSH
- půdní mikrobiologie * MeSH
- reakce na tepelný šok * MeSH
- sinice fyziologie MeSH
- transport elektronů MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- bakteriální proteiny MeSH
- chlorofyl a MeSH
- fotosyntetická reakční centra (proteinové komplexy) MeSH
Cold acclimation modifies the photosynthetic machinery and enables plants to survive at sub-zero temperatures, whereas in warm habitats, many species suffer even at non-freezing temperatures. We have measured chlorophyll a fluorescence (ChlF) and CO2 assimilation to investigate the effects of cold acclimation, and of low temperatures, on a cold-sensitive Arabidopsis thaliana accession C24. Upon excitation with low intensity (40 µmol photons m- 2 s- 1) ~ 620 nm light, slow (minute range) ChlF transients, at ~ 22 °C, showed two waves in the SMT phase (S, semi steady-state; M, maximum; T, terminal steady-state), whereas CO2 assimilation showed a linear increase with time. Low-temperature treatment (down to - 1.5 °C) strongly modulated the SMT phase and stimulated a peak in the CO2 assimilation induction curve. We show that the SMT phase, at ~ 22 °C, was abolished when measured under high actinic irradiance, or when 3-(3, 4-dichlorophenyl)-1, 1- dimethylurea (DCMU, an inhibitor of electron flow) or methyl viologen (MV, a Photosystem I (PSI) electron acceptor) was added to the system. Our data suggest that stimulation of the SMT wave, at low temperatures, has multiple reasons, which may include changes in both photochemical and biochemical reactions leading to modulations in non-photochemical quenching (NPQ) of the excited state of Chl, "state transitions," as well as changes in the rate of cyclic electron flow through PSI. Further, we suggest that cold acclimation, in accession C24, promotes "state transition" and protects photosystems by preventing high excitation pressure during low-temperature exposure.
- Klíčová slova
- 3-(3, 4-dichlorophenyl)-1, 1- dimethylurea, Chlorophyll fluorescence transients, Cold acclimation, Gas-exchange measurements, Low-temperature effect, Methyl viologen, Slow SMT fluorescence phase, State transition,
- MeSH
- aklimatizace MeSH
- Arabidopsis metabolismus MeSH
- chlorofyl a metabolismus MeSH
- fotosyntéza fyziologie MeSH
- nízká teplota MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- chlorofyl a MeSH
