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Govindjee

Publications and source records attributed to Govindjee.

At least 55 records · Page 3Linked to original sources
PubMed1988

Characteristics of Five New Photoautotrophic Suspension Cultures Including Two Amaranthus Species and a Cotton Strain Growing on Ambient CO(2) Levels.

Suspension cultures of cotton (Gossypium hirsutum), Amaranthus cruentus, A. powellii, Datura innoxia, and a Nicotiana tabacum-N. glutinosa fusion hybrid were adapted to grow photoautotrophically under continuous light. The cotton strain grew with an atmosphere of ambient CO(2) (about 0.06 to 0.07% in the culture room) while the other strains required elevated CO(2) levels (5%). Photoautotrophy was indicated by the requirement for CO(2) and for light for growth. The strains grew with doubling times near 14 days and had from 50 to 600 micrograms of chlorophyll per gram of fresh weight. The cells grew in small to moderate sized clumps with cell sizes from 40 to 70 micrometers (diameter). Like most photoautotrophic cultures described so far the ribulose 1,5-bisphosphate carboxylase (RuBPcase) activity levels were well below those of mature leaves. The phosphoenolpyruvate carboxylase levels were not elevated in the C(4)Amaranthus species. The cells showed high dark respiration rates and had lower net CO(2) fixation under high O(2) conditions. Dark CO(2) fixation rates ranged from near 10 to 30% of that in light. Fluorescence emission spectra measurements show that the cell antenna pigments systems of the four strains examined are similar to that of chloroplasts of green plants. The cotton strain which was capable of growth under ambient CO(2) conditions showed the unique properties of a high RuBPcase activation level in ambient CO(2) and a stable ability to show net CO(2) fixation in 21% O(2) conditions.

Journal Article↗
PubMed1986

Bicarbonate, not CO2, is the species required for the stimulation of Photosystem II electron transport.

Evidence is presented that the bicarbonate ion (HCO3-), not CO2, H2CO3 or CO32-, is the species that stimulates electron transport in Photosystem II from spinach (Spinacia oleracea). Advantage was taken of the pH dependence of the ratio of HCO3- to CO2 at equilibrium in order to vary effectively the concentration of one species while holding the other constant. The Hill reaction was stimulated in direct proportion with the equilibrium HCO3- concentration, but it was independent of the equilibrium CO2 concentration. The other two carbonic species, H2CO3 and CO32-, are also shown to have no direct involvement. It is suggested that HCO3- is the species which binds to the effector site.

2,6-Dichloroindophenol↗
PubMed1985

Molecular mechanism of water oxidation in photosynthesis based on the functioning of manganese in two different environments.

We present a model of photosynthetic water oxidation that utilizes the property of higher-valent Mn ions in two different environments and the characteristic function of redox-active ligands to explain all known aspects of electron transfer from H(2)O to Z, the electron donor to P680, the photosystem II reaction center chlorophyll a. There are two major features of this model. (i) The four functional Mn atoms are divided into two groups of two Mn each: [Mn] complexes in a hydrophobic cavity in the intrinsic 34-kDa protein; and (Mn) complexes on the hydrophilic surface of the extrinsic 33-kDa protein. The oxidation of H(2)O is carried out by two [Mn] complexes, and the protons are transferred from a [Mn] complex to a (Mn) complex along the hydrogen bond between their respective ligand H(2)O molecules. (ii) Each of the two [Mn] ions binds one redox-active ligand (RAL), such as a quinone (alternatively, an aromatic amino acid residue). Electron transfer occurs from the reduced RAL to the oxidized Z. When the experimental data concerning atomic structure of the water-oxidizing center (WOC), electron transfer between the WOC and Z, the electronic structure of the WOC, the proton-release pattern, and the effect of Cl(-) are compared with the predictions of the model, satisfactory qualitative and, in many instances, quantitative agreements are obtained. In particular, this model clarifies the origin of the observed absorption-difference spectra, which have the same pattern in all S-state transitions, and of the effect of Cl(-)-depletion on the S states.

Journal Article↗
PubMed1984

NMR study of chloride ion interactions with thylakoid membranes.

The role of Cl(-) in photosynthetic O(2) evolution has been investigated by observing the (35)Cl NMR linewidth under a variety of conditions in aqueous suspensions of chloroplasts, primarily for the halophytes Avicennia germinans, Avicennia marina, and Aster tripolium but also for spinach. The line broadening shows there is weak, ionic binding of Cl(-) to thylakoids, the bound Cl(-) exchanging rapidly (>>10(4) sec(-1)) with free Cl(-) in solution. The binding is necessary for O(2) evolution to occur. Michaelis-Menten constants obtained from the Cl(-) dependence of the O(2) evolution rate are approximately 15-70 mM for the halophytes compared with 0.6 mM for spinach (0.5 mM with Br(-)). There appear to be two types of Cl(-) binding sites in halophytes, of which the stronger is the activator, at lower [Cl(-)], of O(2) evolution. The (35)Cl line broadening includes a nonspecific interaction, which becomes apparent at high Cl(-) concentrations (>/=0.5 M).

Journal Article↗
PubMed1984

Some plant leaves have orientation-dependent EPR and NMR spectra.

Proton nuclear magnetic resonance ((1)H NMR) spectra of leaves from 50 plant species were obtained at a spectrometer frequency of 470 MHz. Water present in leaf samples gives rise to characteristic spectral patterns. Most species show only one broad (1)H NMR peak; however, the leaves of some plants display complex, orientation-dependent spectra in which a common three-line pattern is discerned. The pattern varies with the angle between the leaf surface and the external magnetic field. Proton relaxation measurements show the presence of at least two water compartments in the leaves. The compartments are responsible for different components of the spectral pattern. EPR spectra, obtained at 35 GHz and at a temperature of -180 degrees C, of plant leaf sections are dominated by the strong signals of manganous ions. We find that most plant leaves have isotropic Mn(2+) EPR spectra. However, in some species (including ones that exhibit orientation-dependent (1)H NMR spectra) we detect orientation-dependent intensities in the forbidden lines; the spectra indicate that Mn(2+) ions occupy binding sites with axial or lower symmetry on nonrandomly oriented membranes. Both the NMR and the EPR results suggest that the chloroplasts of some plants are preferentially aligned with respect to the leaf surface.

Journal Article↗
PubMed1984

Charge accumulation and photochemistry in leaves studied by thermoluminescence and delayed light emission.

A major breakthrough in our understanding of how plants oxidize water to molecular O(2) was the discovery by P. Joliot and co-workers that the O(2) yield per flash, in a series of light flashes, oscillates with a periodicity of 4. This led to the concept by B. Kok and co-workers that these reactions involve accumulation of four positive charges in independent "O(2)-evolving centers," which undergo a series of changes in their redox state (the so-called S states). In the present paper, we have applied optical techniques (such as thermoluminescence and delayed light emission, both discovered by W. Arnold and co-workers) to monitor charge storage on the O(2)-evolving system in leaves from higher plants. We observed a period of four oscillations in both thermoluminescence and delayed light emission, with maxima on flashes 2 and 6, establishing a relationship with the charge accumulation process in photosynthesis. These measurements provided additional new information: the deactivation of the "O(2)-evolving centers," which cannot be measured by the O(2) method in the leaves, is in the 20- to 30-s range; and in the dark-adapted leaves, the secondary bound plastoquinone molecule (the so-called secondary electron acceptor Q(B)) is in equal concentration in its reduced and oxidized forms. The origin of thermoluminescence and delayed light emission, in terms of the recombination of charges on the O(2)-evolving and plastoquinone sides, is also discussed.

Journal Article↗
PubMed1983

Energetics of photosynthetic glow peaks.

By postulating temperature-dependent equilibria between two or more electron carriers acting as traps for electrons or holes, it is possible to modify the Randall-Wilkins theory of thermoluminescence so as to explain the abnormally large apparent activation energies and apparent frequency factors observed in photosynthetic glow curves when fitted by unmodified Randall-Wilkins theory. The equilibria serve to inhibit the formation of the light-emitting excited state by withholding the needed precursor state. When the inhibition is released at higher temperature by shift of equilibrium with temperature, the rise of the glow peak can be much faster than would result from Arrhenius behavior based on the true activation energy and so appears to correspond to a higher activation energy accompanied by a larger frequency factor. From another viewpoint, the enthalpy changes, DeltaH, of the equilibria tend to add to the activation energy. Similarly the entropy changes, DeltaS, of the equilibria tend to add to the entropy of activation, giving the large apparent frequency factors. The positive values of DeltaS needed would correspond to entropy decreases in the forward early electron transport. A comparison of the glow peaks obtained by different workers is also presented.

Journal Article↗
PubMed1982

Effects of Cations and Abscisic Acid on Chlorophyll a Fluorescence in Guard Cells of Vicia faba.

The effects of cations and abscisic acid on chloroplast activity in guard cells of Vicia faba were investigated by analysis of the transient of chlorophyll a fluorescence. When epidermal strips containing guard cells as the only living cells were incubated in water and illuminated with strong light, chlorophyll a fluorescence rose rapidly to a high intensity and then declined slowly to a stationary level. The rate of this decline was enhanced by K(+) or Na(+), and the effect of these cations was greater when added with phosphate than with chloride as the anion. Ca(2+) suppressed the enhancement by Na(+) and, to a lesser extent, that by K(+). Abscisic acid also suppressed the enhancement by K(+) and Na(+). Since the fluorescence decline reflects the increase of intrathylakoid H(+) concentration necessary for photophosphorylation, the acceleration of the decline by K(+) (or Na(+) in the absence of Ca(2+)) implicates chloroplast activity in ion accumulation by guard cells in the light. The differential effects of phosphate and chloride suggest that chloroplast activity may be involved in malate formation in guard cells in the light.

Journal Article↗
PubMed1981

On the active species in bicarbonate stimulation of Hill reaction in thylakoid membranes.

At 5 degree C addition of CO2 or HCO3- to CO (HCO3-)-depleted thylakoids (containing 100 mM formate) initiates, within 10 s, the activation of the Hill reaction in light. In contrast to HCO3- addition, where there is a lag of 6-8 s, the activation by CO2 addition is almost instantaneous. With CO2, prior addition of carbonic anhydrase produces a lag of about 6 s that approaches the lag observed by the HCO3- addition. These data suggest that CO2 is an active species involved in stimulating the Hill reaction. Binding of CO2 (HCO3-) to a component on the external side of the thylakoid membranes is supported by the present study.

Bicarbonates↗
PubMed1981

Evidence for a close spatial location of the binding sites for CO2 and for photosystem II inhibitors.

1. CO2-depletion of thylakoid membranes results in a decrease of binding affinity of the Photosystem II (PS II) inhibitor atrazine. The inhibitory efficiency of atrazine, expressed as I50-concentration (50% inhibition) of 2,6-dichlorophenolindophenol reduction, is the same in CO2-depleted as well as in control thylakoids. This shows that CO2-depletion results in a complete inactivation of a part of the total number of electron transport chains. 2. A major site of action of CO2, which had previously been located between the two electron acceptor quinone molecule B (or R) and Photosystem II inhibitor atrazine as suggested by the following observations: (a) CO2-depletion results in a shift of the binding constant (kappa b) of [14C]atrazine to thylakoid membranes indicating a decreased affinity of atrazine to membrane; (b) trypsin treatment, which is known to modify the Photosystem II complex at the level of B, strongly diminishes CO2 stimulation of electron transport reactions in CO2-depleted membranes; and (c) thylakoids from atrazine-resistant plants, which contain a Photosystem II complex modified at the inhibitor binding site, show an altered CO2-stimulation of electron flow. 3. CO2-depletion does not produce structural changes in enzyme complexes involved in Photosystem II function of thylakoid membranes, as shown by freeze-fracture studies using electron microscopy.

Atrazine↗
PubMed1980

Effects of bulk pH and of monovalent and divalent cations on chlorophyll a fluorescence and electron transport in pea thylakoids.

Millimolar concentrations of monovalent cations enhance and divalent cations impede the redistribution (spill-over) of electronic excitation energy from Photosystem (PS) II to PS I in cation-depleted (sucrose-washed) thylakoids; this concept is based on chlorophyll a fluorescence and electron transport measurements over a narrow pH range around 7. We have tested the above concept in pea thylakoids over the pH range 5 to 9 by parallel measurements of various chlorophyll a fluorescence parameters (spectra, transients, and lifetimes at 77 K and 293 K, and polarization at 293 K) and of the rates of partial reactions of PSI and II. Our results provide the following information. (1) Mg2+ enhancement of fluorescence is maximum between 680 and 690 nm and minimum between 710 and 720 nm. (2) The optimum conditions for the observation of the Mg2+-induced enhancement of fluorescence are: wavelength of emission, 685 nm; concentratin of Mg2+, 10 mM, and pH, approximately 7.5. (3) Mg2+ decreases the efficiency of excitation redistribution from PS II to PS I over the pH range 6 to 9. (4) The antagonistic effects between Na+ and Mg2+ hold simultaneously for both the fluorescence intensity and lifetime, at physiological temperatures, only within the pH range 6 to 8. (5) Mg2+ enhances the light-limited electron transport rate through PS II in the pH range 5.4 to 8.2 and decreases that through PS I at pH 7.1 and 8.2. The % increase in PS II is, however, about twice the % decrease in PS I.

Cations↗
PubMed1979

Conformation and activity of chloroplast coupling factor exposed to low chemical potential of water in cells.

(1) Photophosphorylation, Ca2+-ATPase and Mg2+-ATPase activities of isolated chloroplasts were inhibited 55--65% when the chemical potential of water was decreased by dehydrating leaves to water potentials (psi w) of --25 bars before isolation of the plastids. The inhibition could be reversed in vivo by rehydrating the leaves. (2) These losses in activity were reflected in coupling factor (CF1) isolated from the leaves, since CF1 from leaves with low psi w had less Ca2+-ATPase activity than control CF1 and did not recouple phosphorylation in CF1-deficient chloroplasts. In contrast, CF1 from leaves having high psi w only partially recoupled phosphorylation by CF1-deficient chloroplasts from leaves havig low psi w. This indicated that low psi w affected chloroplast membranes as well as CF1 itself. (3) Coupling factor from leaves having low psi w had the same number of subunits, and the same electrophoretic mobility, and could be obtained with the same yields as CF1 from control leaves. However, direct measurements of fluorescence polarization, ultraviolet absorption, and circular dichroism showed that CF1 from leaves having low psi w differed from control CF1. The CF1 from leaves having low psi w also had decreased ability to bind fluorescent nucleotides (epsilon-ATP and epsilon-ADP). (4) Exposure of isolated CF1 to low psi w in vitro by preincubation in sucrose-containing media inhibited the Ca2+-ATPase activity of the protein in subsequent assays without sucrose. Inclusion of 5 or 10 mM Mg2+ in the preincubation medium markedly inhibited Ca2+-ATPase activity. (5) These results show that CF1 undergoes changes in cells which alter its phosphorylating ability. Since low cell psi w changed the spectroscopic properties but not other protein properties of CF1, the changes were most likely caused by altered confurn, photophosphorylation. The inhibition of ATPase activity in CF1 in vitro at low psi w and high ion concentration mimicked the change in activity seen in vivo.

Adenosine Triphosphatases↗
PubMed1978

Nuclear magnetic relaxation by the manganese in aqueous suspensions of chloroplasts.

Proton and oxygen-17 NMR relaxation rate (T1-1 and T2-1) data are presented for aqueous suspensions of dark-adapted chloroplasts. It is concluded from the dependence of the proton relaxation rates (PRR) upon Mn concentration that T1-1 and T2-1 are determined largely by the loosely bound Mn present in the chloroplast membranes. The frequency and temperature dependences of PRR are characteristic of Mn(II). The effects of oxidants (e.g., ferricyanide) and reductants (e.g., tetraphenylboron) on the PRR indicate that only about one-third to one-fourth of the loosely bound Mn is present in the dark-adapted chloroplasts as Mn(II), the remainder being in a higher oxidation state(s), probably Mn(III). The frequency dependence of the PRR for the chloroplast suspensions was fitted by a simplified form of the Solomon-Bloembergen-Morgan equations, and the following parameters were obtained: tauS = (1.1 +/- 0.1) X 10(-8) S; tauM = (2.2 +/- 0.2) X 10(-8) S; and B = (0.9 +/- 0.09) X 10(19). The oxygen-17 T1 and T2 data for suspensions before and after treatment with a detergent are consistent with the location of the manganese in the interior of the thylakoids. An analysis of the relaxation rates shows that the average lifetime of a water molecule inside a thylakoid is greater than 1 ms.

Chloroplasts↗