Excitation transfer and quenching in photosystem II, enlightened by carotenoid triplet state in leaves.

Accumulation of carotenoid (Car) triplet states was investigated by singlet-triplet annihilation, measured as chlorophyll (Chl) fluorescence quenching in sunflower and lettuce leaves. The leaves were illuminated by Xe flashes of 4 μs length at half-height and 525-565 or 410-490 nm spectral band, maximum intensity 2 mol quanta m s, flash photon dose up to 10 μmol m or 4-10 PSII excitations. Superimposed upon the non-photochemically unquenched F state, fluorescence was strongly quenched near the flash maximum (minimum yield F), but returned to the F level after 30-50 μs.

VHL-deficiency leads to reductive stress in renal cells.

Heritable renal cancer syndromes (RCS) are associated with numerous chromosomal alterations including inactivating mutations in von Hippel-Lindau (VHL) gene. Here we identify a novel aspect of the phenotype in VHL-deficient human renal cells. We call it reductive stress as it is characterised by increased NADH/NAD ratio that is associated with impaired cellular respiration, impaired CAC activity, upregulation of reductive carboxylation of glutamine and accumulation of lipid droplets in VHL-deficient cells.

Prying into the green black-box.

Life-long efforts of the Tartu photosynthesis research group have been summarized. The measurements were facilitated by self-designed instruments, distinct in multifunctionality and fastresponse time. The black-box type kinetical analysis on intact leaves has revealed several physiologically significant features of leaf photosynthesis.

Hypothermia Alleviates Reductive Stress, a Root Cause of Ischemia Reperfusion Injury.

Ischemia reperfusion injury is common in transplantation. Previous studies have shown that cooling can protect against hypoxic injury. To date, the protective effects of hypothermia have been largely associated with metabolic suppression.

Time- and reduction-dependent rise of photosystem II fluorescence during microseconds-long inductions in leaves.

Lettuce (Lactuca sativa) and benth (Nicotiana benthamiana) leaves were illuminated with 720 nm background light to mix S-states and oxidize electron carriers. Green-filtered xenon flashes of different photon dose were applied and O evolution induced by a flash was measured. After light intensity gradient across the leaf was mathematically considered, the flash-induced PSII electron transport (= 4·O evolution) exponentially increased with the flash photon dose in any differential layer of the leaf optical density.

Variable fluorescence of closed photochemical reaction centers.

Chlorophyll fluorescence induction during 0.4 to 200 ms multiple-turnover pulses (MTP) was measured in parallel with O evolution induced by the MTP light. Additionally, a saturating single-turnover flash (STF) was applied at the end of each MTP and the total MTP +STF O evolution was measured.

Kinetics of photosystem II electron transport: a mathematical analysis based on chlorophyll fluorescence induction.

The OJDIP rise in chlorophyll fluorescence during induction at different light intensities was mathematically modeled using 24 master equations describing electron transport through photosystem II (PSII) plus ordinary differential equations for electron budgets in plastoquinone, cytochrome f, plastocyanin, photosystem I, and ferredoxin. A novel feature of the model is consideration of electron in- and outflow budgets resulting in changes in redox states of Tyrosine Z, P680, and Q as sole bases for changes in fluorescence yield during the transient. Ad hoc contributions by transmembrane electric fields, protein conformational changes, or other putative quenching species were unnecessary to account for primary features of the phenomenon, except a peculiar slowdown of intra-PSII electron transport during induction at low light intensities.

Kinetics of plastoquinol oxidation by the Q-cycle in leaves.

Electrochromic shift measurements confirmed that the Q-cycle operated in sunflower leaves. The slow temporarily increasing post-pulse phase was recorded, when ATP synthase was inactivated in the dark and plastoquinol (PQH(2)) oxidation was initiated by a short pulse of far-red light (FRL). During illumination by red light, the Q-cycle-supported proton arrival at the lumen and departure via ATP synthase were simultaneous, precluding extreme build-up of the membrane potential.

Oxidation of plastohydroquinone by photosystem II and by dioxygen in leaves.

In sunflower leaves linear electron flow LEF=4O2 evolution rate was measured at 20 ppm O2 in N2. PSII charge separation rate CSRII=aII∙PAD∙(Fm-F)/Fm, where aII is excitation partitioning to PSII, PAD is photon absorption density, Fm and F are maximum and actual fluorescence yields. Under 630 nm LED+720 nm far-red light (FRL), LEF was equal to CSRII with aII=0.

Fluorescence F 0 of photosystems II and I in developing C3 and C 4 leaves, and implications on regulation of excitation balance.

This work addresses the question of occurrence and function of photosystem II (PSII) in bundle sheath (BS) cells of leaves possessing NADP-malic enzyme-type C4 photosynthesis (Zea mays). Although no requirement for PSII activity in the BS has been established, several component proteins of PSII have been detected in BS cells of developing maize leaves exhibiting O2-insensitive photosynthesis. We used the basal fluorescence emissions of PSI (F 0I) and PSII (F 0II) as quantitative indicators of the respective relative photosystem densities.

Action spectra of photosystems II and I and quantum yield of photosynthesis in leaves in State 1.

The spectral global quantum yield (YII, electrons/photons absorbed) of photosystem II (PSII) was measured in sunflower leaves in State 1 using monochromatic light. The global quantum yield of PSI (YI) was measured using low-intensity monochromatic light flashes and the associated transmittance change at 810nm. The 810-nm signal change was calibrated based on the number of electrons generated by PSII during the flash (4·O2 evolution) which arrived at the PSI donor side after a delay of 2ms.

Competition between isoprene emission and pigment synthesis during leaf development in aspen.

In growing leaves, lack of isoprene synthase (IspS) is considered responsible for delayed isoprene emission, but competition for dimethylallyl diphosphate (DMADP), the substrate for both isoprene synthesis and prenyltransferase reactions in photosynthetic pigment and phytohormone synthesis, can also play a role. We used a kinetic approach based on post-illumination isoprene decay and modelling DMADP consumption to estimate in vivo kinetic characteristics of IspS and prenyltransferase reactions, and to determine the share of DMADP use by different processes through leaf development in Populus tremula. Pigment synthesis rate was also estimated from pigment accumulation data and distribution of DMADP use from isoprene emission changes due to alendronate, a selective inhibitor of prenyltransferases.

Thermal phase and excitonic connectivity in fluorescence induction.

Chl fluorescence induction (FI) was recorded in sunflower leaves pre-adapted to darkness or low preferentially PSI light, or inhibited by DCMU. For analysis the FI curves were plotted against the cumulative number of excitations quenched by PSII, n q, calculated as the cumulative complementary area above the FI curve. In the +DCMU leaves n q was <1 per PSII, suggesting pre-reduction of Q A during the dark pre-exposure.

Photosystem II antennae are not energetically connected: evidence based on flash-induced O2 evolution and chlorophyll fluorescence in sunflower leaves.

Oxygen evolution was measured in sunflower leaves in steady-state and during multiple-turnover pulses (MTP) of different light (630 nm LED plus far-red light) intensity and duration. In parallel, Chl fluorescence yields F(0) (minimum), F(s) (steady-state), and F(m) (pulse-saturated), as well as fluorescence induction during MTPs were recorded. Extra O(2) evolution was measured in response to a saturating single-turnover Xe flash (STF) applied immediately subsequently to the actinic light in the steady-state and to each MTP.

Oxygen evolution and chlorophyll fluorescence from multiple turnover light pulses: charge recombination in photosystem II in sunflower leaves.

Oxygen evolution and Chl fluorescence induction were measured during multiple turnover light pulses (MTP) of 630-nm wavelength, intensities from 250 to 8,000 μmol quanta m(-2) s(-1) and duration from 0.3 to 200 ms in sunflower leaves at 22 °C. The ambient O(2) concentration was 10-30 ppm and MTP were applied after pre-illumination under far-red light (FRL), which oxidized plastoquinone (PQ) and randomized S-states because of the partial excitation of PSII.

Oxygen evolution from single- and multiple-turnover light pulses: temporal kinetics of electron transport through PSII in sunflower leaves.

Oxygen evolution per single-turnover flash (STF) or multiple-turnover pulse (MTP) was measured with a zirconium O(2) analyzer from sunflower leaves at 22 °C. STF were generated by Xe arc lamp, MTP by red LED light of up to 18000 μmol quanta m(-2) s(-1). Ambient O(2) concentration was 10-30 ppm, STF and MTP were superimposed on far-red background light in order to oxidize plastoquinone (PQ) and randomize S-states.

The size of the lumenal proton pool in leaves during induction and steady-state photosynthesis.

This report describes a new method to measure the chloroplastic lumenal proton pool in leaves (tobacco and sunflower). The method is based on measurement of CO(2) outbursts from leaves caused by the shift in the CO(2) + H(2)O ↔ HCO(3)(-) + H(+) equilibrium in the chloroplast stroma as protons return from the lumen after darkening. Protons did not accumulate in the lumen to a significant extent when photosynthesis was light-limited, but a large pool of >100 μmol H(+) m(-2) accumulated in the lumen as photosynthesis became light-saturated.

Induction of a longer term component of isoprene release in darkened aspen leaves: origin and regulation under different environmental conditions.

After darkening, isoprene emission continues for 20 to 30 min following biphasic kinetics. The initial dark release of isoprene (postillumination emission), for 200 to 300 s, occurs mainly at the expense of its immediate substrate, dimethylallyldiphosphate (DMADP), but the origin and controls of the secondary burst of isoprene release (dark-induced emission) between approximately 300 and 1,500 s, are not entirely understood. We used a fast-response gas-exchange system to characterize the controls of dark-induced isoprene emission by light, temperature, and CO(2) and oxygen concentrations preceding leaf darkening and the effects of short light pulses and changing gas concentrations during dark-induced isoprene release in hybrid aspen (Populus tremula × Populus tremuloides).

Temperature response of isoprene emission in vivo reflects a combined effect of substrate limitations and isoprene synthase activity: a kinetic analysis.

The responses of isoprene emission rate to temperature are characterized by complex time-dependent behaviors that are currently not entirely understood. To gain insight into the temperature dependencies of isoprene emission, we studied steady-state and transient responses of isoprene emission from hybrid aspen (Populus tremula × Populus tremuloides) leaves using a fast-response gas-exchange system coupled to a proton-transfer reaction mass spectrometer. A method based on postillumination isoprene release after rapid temperature transients was developed to determine the rate constant of isoprene synthase (IspS), the pool size of its substrate dimethylallyldiphosphate (DMADP), and to separate the component processes of the temperature dependence of isoprene emission.

Equilibrium or disequilibrium? A dual-wavelength investigation of photosystem I donors.

Oxidation of photosystem I (PSI) donors under far-red light (FRL), slow re-reduction by stromal reductants and fast re-reduction in the dark subsequent to illumination by white light (WL) were recorded in leaves of several C(3) plants at 810 and 950 nm. During the re-reduction from stromal reductants the mutual interdependence of the two signals followed the theoretical relationship calculated assuming redox equilibrium between plastocyanin (PC) and P700, with the equilibrium constant of 40 +/- 10 (Delta E (m) = 86-99 mV) in most of the measured 24 leaves of nine plant species. The presence of non-oxidizable PC of up to 13% of the whole pool, indicating partial control of electron transport by PC diffusion, was transiently detected during a saturation pulse of white light superimposed on FRL or on low WL.

Fast cyclic electron transport around photosystem I in leaves under far-red light: a proton-uncoupled pathway?

Fast cyclic electron transport (CET) around photosystem I (PS I) was observed in sunflower (Helianthus annuus L.) leaves under intense far-red light (FRL) of up to 200 mumol quanta m(-2) s(-1). The electron transport rate (ETR) through PS I was found from the FRL-dark transmittance change at 810 and 950 nm, which was deconvoluted into redox states and pool sizes of P700, plastocyanin (PC) and cytochrome f (Cyt f).

Evidence that light, carbon dioxide, and oxygen dependencies of leaf isoprene emission are driven by energy status in hybrid aspen.

Leaf isoprene emission scales positively with light intensity, is inhibited by high carbon dioxide (CO(2)) concentrations, and may be enhanced or inhibited by low oxygen (O(2)) concentrations, but the mechanisms of environmental regulation of isoprene emission are still not fully understood. Emission controls by isoprene synthase, availability of carbon intermediates, or energetic cofactors have been suggested previously. In this study, we asked whether the short-term (tens of minutes) environmental control of isoprene synthesis results from alterations in the immediate isoprene precursor dimethylallyldiphosphate (DMADP) pool size, and to what extent DMADP concentrations are affected by the supply of carbon and energetic metabolites.

Postillumination isoprene emission: in vivo measurements of dimethylallyldiphosphate pool size and isoprene synthase kinetics in aspen leaves.

The control of foliar isoprene emission is shared between the activity of isoprene synthase, the terminal enzyme catalyzing isoprene formation from dimethylallyldiphosphate (DMADP), and the pool size of DMADP. Due to limited in vivo information of isoprene synthase kinetic characteristics and DMADP pool sizes, the relative importance of these controls is under debate. In this study, the phenomenon of postillumination isoprene release was employed to develop an in vivo method for estimation of the DMADP pool size and to determine isoprene synthase kinetic characteristics in hybrid aspen (Populus tremula x Populus tremuloides) leaves.

Kinetics of leaf oxygen uptake represent in planta activities of respiratory electron transport and terminal oxidases.

We present, for the first time, the oxygen response kinetics of mitochondrial respiration measured in intact leaves (sunflower and aspen). Low O(2) concentrations in N(2) (9-1500 ppm) were preset in a flow-through gas exchange measurement system, and the decrease in O(2) concentration and the increase in CO(2) concentration as result of leaf respiration were measured by a zirconium cell O(2) analyser and infrared-absorption CO(2) analyser, respectively. The low O(2) concentrations little influenced the rate of CO(2) evolution during the 60-s exposure.

Rates and roles of cyclic and alternative electron flow in potato leaves.

Measurements of 810 nm transmittance changes in leaves, simultaneously with Chl fluorescence, CO(2) uptake and O(2) evolution, were carried out on potato (Solanum tuberosum L.) leaves with altered expression of plastidic NADP-dependent malate dehydrogenase. Electron transport rates were calculated: J(C) from the CO(2) uptake rate considering ribulose-1,5-bisphosphate (RuBP) carboxylation and oxygenation, J(O) from the O(2) evolution rate, J(F) from Chl fluorescence parameters and J(I) from the post-illumination re-reduction speed of PSI donors.