Photovoltaic effect in pigment films in contact with an electrolytes. V. Theory of the Becquerel photovoltaic effect in porous films of phthalocyanine]

In accordance with literature and our experimental data a theoretical analysis of the model system metal-porous film of the pigment-electrolyte was carried out at stationary illumination in the regimes of photocurrent and photopotential. The main properties and positions of the model are supported experimentally. Specific behaviour of the photovoltaic system considered resides in the discovered dependence of the transfer mechanism on the value of catode potential of the electrode.     

Reversible inhibition of proton release activity and the anesthetic-induced acid-base equilibrium between the 480 and 570 nm forms of bacteriorhodopsin.

In purple membrane added with general anesthetics, there exists an acid-base equilibrium between two spectral forms of the pigment: bR570 and bR480 (apparent pKa = 7.3). As the purple 570 nm bacteriorhodopsin is reversibly transformed into its red 480 nm form, the proton pumping capability of the pigment reversibly decreases, as indicated by transient proton release measurements and proton translocation action spectra of mixture of both spectral forms. It happens in spite of a complete photochemical activity in bR480 that is mostly characterized by fast deprotonation and slow reprotonation steps and which, under continuous illumination, bleaches with a yield comparable to that of bR570. This modified photochemical activity has a correlated specific photoelectrical counterpart: a faster proton extrusion current and a slower reprotonation current. The relative areas of all photocurrent phases are reduced in bR480, most likely because its photochemistry is accompanied by charge movements for shorter distances than in the native pigment, reflecting a reversible inhibition of the pumping activity.     

Microspectrophotometric evidence for two photointerconvertible states of visual pigment in the barnacle lateral eye

Microspectrophotometrically derived difference spectra from the barnacles Balanus amphitrite and B. eburneus show that a blue illumination after an orange illumination causes a decrease in absorption in the blue region and an increase in absorption in the green-yellow region, with an isosbestic point around 535 nm. Orange-following-blue illumination causes the reverse changes. The dark time between the adapting and measuring lights has no influence on the data. The results confirm previously reported ERP measurements which indicate that the barnacle visual pigment has two photointerconvertible dark-stable states. If one assumes a Dartnall nomogram shape for the two absorption spectra, a best fit to the observed difference spectra is obtained with nomograms peaking at 492 nm and 532 nm, with a peak absorbance ratio around 1.6:1. These two nomograms fit very well the ERP action spectra of metarhodopsin and rhodopsin, respectively, thus indicating that the ERP is a reliable measure of visual-pigment changes in the barnacle. The existence of a photostable blue pigment is demonstrated in B. eburneus and in some of B. amphitrite receptors, and the possible influence of this photostable pigment on the various action spectra measured in the barnacle is discussed.     

Effects of light on pigment development in the echiuran worm, Bonellia viridis

In rhe echiuran Bonellia viridis the green pigment first develops in the gastrula, before the formation of the trochal rings, and the trochophores acquire a deep green pigmentation before hatching. Eggs that were cultured under continuous illumination (2000–4000 lx) did not develop any green pigment, and although the trochophores hatched normally, they lacked pigment entirely. The unpigmented trochophores that were kept under continuous illumination developed into unpigmented males and females. The far-blue (380–400 nm) and red (620–640 nm) regions were more effective in preventing pigment development than the blue, green and yellow regions of the spectrum. Illumination of the eggs before the gastrula stage did not affect the development of pigment when these were subsequently returned to the dark. When unpigmented trochophores, that had been cultured in the light, were placed in the dark they acquired a slight pigmentation, but this was much less intense than that of trochophores that had been cultured in the dark throughout. Pigmented trochophores were more vulnerable to high light intensities than unpigmented ones.     

Insect pupil mechanisms

Summary The light-dependent pigment migration system of dragonfly ocelli was studied by optical, non-invasive techniques. The median ocellus is comprised of two lateral halves, as can be demonstrated in the intact animal since illumination of the receptors in one half of the median ocellus only induces a movement of pigment located in that half. Measurable pigment migration can occur within a few seconds, but its speed and extent depends on light intensity. Dispersal of pigment, which occurs upon light adaptation, proceeds faster than retraction, which occurs upon dark adaptation. Action spectra for pigment movement have been determined inSympetrum andAnax. The spectrum forSympetrum has a prominent UV peak, moderate blue sensitivity, and very low green sensitivity. A similar profile is obtained inAnax, but only after intense orange adaptation which suppresses the green sensitivity. The results conform to the known spectral sensitivities of Libellulid and Aeschnid ocellar receptors. It is concluded that the photoreceptors drive pigment movement through an unknown mechanism. The effect of the migration of pigment is the selective reduction of radiant flux on the retina from luminous sources at high elevations relative to the animal's normal flying posture.This work was supported by grants by the Netherlands Organization for the Advancement of Pure Research (Z.W.O.) (to DGS), by National Eye Institute U.S.P.H.S. EY01140 and EY00785 (to GDB) and EY00777 and EY00040 (to RLC).     

The kinetics of visual pigment systems

Using early receptor potential (ERP) measurements, we show that the bistable pigment in the barnacle photoreceptor behaves according to the conclusions of the preceding article (Hochstein et al., 1978): (1) The populations of both stable states approach their steady-state or saturation values under steady illumination exponentially with the same rate constant; the wavelength dependence of this rate constant is called the relaxation spectrum. — (2) The saturation values are independent of initial population and of light intensity; the wavelength dependence of the saturation population is called the saturation spectrum. — (3) The measured relaxation and saturation spectra agree with those calculated, by the theory of the preceding article, from the experimentally determined transition parameters of the pigment system. — We then demonstrate the applicability of relaxation and saturation measurements to the question of whether a single bi-stable pigment system serves, or two or more separate systems serve, as the origins(s) of the ERP and of other phenomena observed in the barnacle photoreceptor: The prolonged depolarizing afterpotential (PDA) and its depression and prevention (anti-PDA). By showing that the relaxation spectra for these phenomena match one another and that of the ERP, and that the same is true for the saturation spectra, we demonstrate that these phenomena originate from the same single bi-stable pigment system as the ERP.     

Multiple light-induced reactions of cytochromes b and c in Rhodopseudomonas spheroides

Illumination of chromatophore preparations from Rhodopseudomonas spheroides causes the oxidation of a cytochrome c and a slight oxidation of a cytochrome b with a maximum at 560nm. When illuminated in the presence of antimycin A the oxidation of cytochrome c was more pronounced and cytochrome b(560) was reduced; the dark oxidation of cytochrome b(560) was biphasic in the presence of succinate, but not in the presence of NADH, a less effective reductant. Split-beam spectroscopy showed that, in addition to the reduction of cytochrome b(560), another pigment with maxima at 565 and 537nm. was reduced and was more rapidly oxidized in the dark than cytochrome b(560). This pigment, tentatively identified as cytochrome b(565), was also detected in spectra at 77 degrees k, after brief illumination at room temperature; the maxima at 77 degrees k were at 562 and 536nm. In the absence of antimycin A, light caused a transient reduction of cytochrome b(565) and an oxidation of cytochrome b(560). Dark oxidation of b(565) was rapid, even in the presence of antimycin A and succinate. Difference spectra, at 77 degrees k, of ascorbate-reduced minus succinate-reduced chromatophores or of anaerobic succinate-reduced minus aerobic succinate-reduced chromatophores suggested that two cytochromes c were present, with maxima at 547 and 549nm. When chromatophores frozen at 77 degrees k were illuminated both these cytochromes c were oxidized, indicating a close association with the photochemical reaction centre. A scheme involving two reaction centres is proposed to explain these results.     

Photo-induced destruction of DOPA-melanin

Studies of the effect of illumination at different wavelengths revealed a high stability of DOPA-melanin to the visible light. Contrariwise, UV-visible light of high intensity upon prolonged (many hours) illumination caused a significant bleaching of the diluted aqueous solution of the pigment. The absorption spectrum of DOPA-melanin in the UV- and IR-regions did not differ from the initial one; however, the illuminated pigment acquired an ability for fluorescence. Stepwise gel chromatography on Toyopearl-55 and Toyopearl-40 columns resulted in three fractions of photobleached DOPA-melanin which differed in their molecular masses, absorption spectra (UV, IR) and fluorescence. It was concluded that the photoinduced bleaching of DOPA-melanin was mainly due to the pigment depolymerization. A possible mechanism of melanin photodestruction is discussed.     

Photoelectric properties of chlorophyll and carotene solutions in nematic liquid crystal located between semiconducting electrodes

The photopotential and photocurrent generation for chlorophyll a, beta-carotene and a mixture of these pigments dissolved in nematic liquid crystal and located between transparent semiconducting electrodes were measured. Both pigments exhibit photopotential and photocurrent generation. From the photocurrent amplitudes it follows that the efficiency of electron transfer to a semiconducting electrode from beta-carotene is higher than from chlorophyll alpha. The photocurrent amplitude of the pigment mixture is slightly lower than that calculated as a sum of amplitudes of pigments located in separated cells. This difference can be explained by secondary effects, such as competition between carotene and chlorophyll molecules in a process of adsorption on a semiconducting electrode. Therefore it seems that no charge transfer complexes of chlorophyll and carotene are formed in the investigated model system.     

Photodamage to Pigment in the Photosystem Ⅱ Reaction Center D1/D2/Cytochrome b559 Complex

Strong light (800 μmol photons/m2 per s)-induced bleaching of the pigment in the isolated photosystem Ⅱ reaction center (PSII RC) under aerobic conditions (in the absence of electron donors or acceptors) was studied using high-pressure liquid chromatography (HPLC), absorption spectra, 77K fluorescence spectra and resonance Raman spectra. Changes in pigment composition of the PSll RC as determined by HPLC after light treatment were as follows: with increasing illumination time chlorophyll (Chi) a and β-carotene (β-car)content decreased. However, decreases in pheophytin (Pheo) could not be observed because of the mixture of the Pheo formed by degraded chlorophyll possibly. On the basis of absorption spectra, it was determined that, with a short time of illumination, the initial bleaching occurred maximally at 680 nm but that with increasing illumination time there was a blue shift to 678 nm. It was suggested that P680 was destroyed initially, followed by the accessory chlorophyll. The activity of P680 was almost lost after 10 min light treatment. Moreover, the bleaching of Pheo and β-car was observed at the beginning of illumination.After illumination, the fluorescence emission intensity changed and the fluorescence maximum blue shifted,showing that energy transfer was disturbed. Resonance Raman spectra of the PSII RC excited at 488.0 and 514.5 nm showed four main bands, peaking at 1 527 cm-1 (υ1), 1 159 cm-1 (υ2), 1 006 cm-1 (υ3), 966 cm-1 (υ4) for 488.0 nm excitation and 1 525 cm-1 (υ1), 1 159 cm-1 (υ2), 1 007 cm-1 (υ3), 968 cm-1 (υ4) for 514.5 nm excitation.It was confirmed that two spectroscopically different β-car molecules exist in the PSII RC. After light treatment for 20 min, band positions and bandwidths were unchanged. This indicates that carotenoid configuration is not the parameter that regulates photoprotection in the PSII RC.     

Light-driven primary sodium ion transport in halobacterium halobium membranes

Light-induced sodium extrusion from H halobium cell envelope vesicles proceeds largely through an uncoupler-sensitive pathway involving bacteriorhodopsin and a proton/sodium antiporter. Vesicles from bacteriorhodopsin-negative strains also extrude sodium ions during illumination, but this transport is not sensitive to uncouplers and has been proposed to involve a light-energized primary sodium pump. Proton uptake in such vesicles is passive, and under steady-state illumination the large electrical potential (negative inside) is just balanced by a pH difference (acid inside), so that the protonmotive force is near zero. Action spectra indicated that this effect of illumination is attributable to a pigment absorbing near 585 nm (of 568 for bacteriorhodopsin). Bleaching of the vesicles by prolonged illumination with hydroxylamine results in inactivation of the transport; retinal addition causes partial return of the activity. Retinal addition also causes the appearance of an absorption peak at 588 nm, while the absorption of free retinal decreases. The 588 nm pigment is present in very small quantities (0.13 nmole/mg protein), and behaves differently from bacteriorhodopsin in a number of respects. Vesicles can be prepared from bacteriorhodopsin-containing H halobium strains in which primary transport for both protons and sodium can be observed. Both pumps appear to cause the outward transport of the cations. The observations indicate the existence of a second retinal protein, in addition to bacteriorhodopsin, in H halobium, which is associated with primary sodium translocation. The initial proton uptake normally observed during illumination of whole H halobium cells may therefore be a passive flux in response to the primary sodium extrusion.     

Antagonistic Components of the Late Receptor Potential in the Barnacle Photoreceptor Arising from Different Stages of the Pigment Process

The late receptor potential (LRP) recorded in barnacle photoreceptor cells exhibits, at high light levels, a strong dependence on the color of the stimulus and of the preceding adaptation. Most strikingly, red illumination of a cell previously adapted to blue light results in a depolarization which may last for up to 30 min after the light goes off, while blue illumination of a cell previously adapted to red light cuts short this extended depolarization or prevents its induction by a closely following red light. Comparison of the action spectra for the stimulus-coincident LRP and for the extended depolarization and its curtailment with those previously measured for the early receptor potential (ERP) confirms that these phenomena derive from the same bi-stable pigment as the ERP. The stimulus-coincident response and the extended depolarization appear to arise from substantial activation of the stable 532 nm state of the pigment, while activation of the stable 495 state depresses or prevents the extended depolarization and probably also depresses the stimulus-coincident response. Since either process can precede the other, with mutually antagonistic effects, one is not simply the reversal of the other; they must be based on separate mechanisms. Furthermore, comparison with ERP kinetics shows that both processes involve mechanisms additional to the pigment changes, as seen in the ERP. A model is proposed and discussed for the LRP phenomena and their dependences on wavelength, intensity, and duration of illumination based on excitor-inhibitor interactions.     

The carotenoid shift in Rhodopseudomonas sphaeroides. The flash induced change

A mutant, Rhodopseudomonas sphaeroides G1C, having only one major carotenoid, neurosporene, is described. The spectrum of the carotenoid shift in this mutant is analysed and it is concluded that only 7–11% of the pigment is involved under conditions of steady-state illumination and that this pigment undergoes a shift of 7 nm.The spectrum of the carotenoid shift under conditions of multi-flash illumination is examined for changes in shape concordant with a progressive red shift of the pigment with increasing membrane potential; the spectra of the fast change after each of three flashes does not agree well with predictions from a model involving a progressive shift of the pigment, the slow change shows qualitative agreement with such a model but the small size of the signal and the presence of more than one phase makes analysis of this phase more difficult.No separate pool of carotenoid, that might correspond to that postulated to participate in the carotenoid shift, could be identified by fourth derivative analysis of, or curve fitting to, the spectrum of the neurosporene.     

The carotenoid shift in Rhodopseudomonas sphaeroides. The flash induced change.

A mutant, Rhodopseudomonas sphaeroides GIC, having only one major carotenoid, neurosporene, is described. The spectrum of the carotenoid shift in this mutant is analysed and it is concluded that only 7-11% of the pigment is involved under conditions of steady-state illumination and that this pigment undergoes a shift of 7 nm. The spectrum of the carotenoid shift under conditions of multi-flash illumination is examined for changes in shape concordant with a progressive red shift of the pigment with increasing membrane potential; the spectra of the fast change after each of three flashes does not agree well with predictions from a model involving a progressive shift of the pigment, the slow change shows qualitative agreement with such a model but the small size of the signal and the presence of more than one phase makes analysis of this phase more difficult. No separate pool of carotenoid, that might correspond to that postulated to participate in the carotenoid shift, could be identified by fourth derivative analysis of, or curve fitting to, the spectrum of the neurosporene.     

Microspectrophotometric characterization and localization of three visual pigments in the compound eye ofNotonecta glauca L. (Heteroptera)

Summary The absorption maxima ( _max) of the visual pigments in the ommatidia ofNotonecta glauca were found by measuring the difference spectra of single rhabdomeres after alternating illumination with two different adaptation wavelengths. All the peripheral rhabdomeres contain a pigment with an extinction maximum at 560 nm. This pigment is sensitive to red light up to wavelengths > 700 nm. In a given ommatidium in the dorsal region of the eye, the two central rhabdomeres both contain one of two pigments, either a pigment with an absorption maximum in the UV, at 345 nm, or — in neighboring rhabdoms — a pigment with an absorption maximum at 445 nm. In the ventral part of the eye only the pigment absorbing maximally in the UV was found in the central rhabdomeres. The spectral absorption properties of various types of screening-pigment granules were measured.     

Photodamage to Pigment in the Photosystem Ⅱ Reaction Center D1/D2/Cytochrome b559 Complex

Strong light （800μmol photons/m^2 per s）-induced bleaching of the pigment in the isolated photosystem Ⅱ reaction center （PSII RC） under aerobic conditions （in the absence of electron donors or acceptors） was studied using high-pressure liquid chromatography （HPLC）, absorption spectra, 77K fluorescence spectra and resonance Raman spectra. Changes in pigment composition of the PSII RC as determined by HPLC after light treatment were as follows： with Increasing illumination time chlorophyll （Chl） a and β-carotene （β-car） content decreased. However, decreases in pheophytin （Pheo） could not be observed because of the mixture of the Pheo formed by degraded chlorophyll possibly. On the basis of absorption spectra, it was determined that, with a short time of illuminatlon, the initial bleaching occurred maximally at 680 nm but that with Increasing Illumination time there was a blue shift to 678 nm. It was suggested that P680 was destroyed Initially, followed by the accessory chlorophyll. The activity of P680 was almost lost after 10 mln light treatment. Moreover, the bleaching of Pheo and β-car was observed at the beginning of illumination. After Illumination, the fluorescence emission Intensity changed and the fluorescence maximum blue shifted, showing that energy transfer was disturbed. Resonance Raman spectra of the PSII RC excited at 488.0 and 514.5 nm showed four main bands, peaking at 1 527 cm^-1 （υ101）, 1 159 cm^-1 （υ2）, 1 006 cm^-1 （υ3）, 966 cm^-1 （υ4） for 488.0 nm excitation and 1 525 cm^-1 （υ1）, 1 159 cm^-1 （υ2）, 1 007 cm^-1 （υ3）, 968 cm^-1 （υ4） for 514.5 nm excitation. It was confirmed that two spectroscopically different β-car molecules exist In the PSII RC. After light treatment for 20 mln, band positions and bandwidths were unchanged. This indicates that carotenoid configuration Is not the parameter that regulates photoprotectlon in the PSII RC.     

Metarhodopsin control by arrestin,light-filtering screening pigments,and visual pigment turnover in invertebrate microvillar photoreceptors

The visual pigments of most invertebrate photoreceptors have two thermostable photo-interconvertible states, the ground state rhodopsin and photo-activated metarhodopsin, which triggers the phototransduction cascade until it binds arrestin. The ratio of the two states in photoequilibrium is determined by their absorbance spectra and the effective spectral distribution of illumination. Calculations indicate that metarhodopsin levels in fly photoreceptors are maintained below ~35% in normal diurnal environments, due to the combination of a blue-green rhodopsin, an orange-absorbing metarhodopsin and red transparent screening pigments. Slow metarhodopsin degradation and rhodopsin regeneration processes further subserve visual pigment maintenance. In most insect eyes, where the majority of photoreceptors have green-absorbing rhodopsins and blue-absorbing metarhodopsins, natural illuminants are predicted to create metarhodopsin levels greater than 60% at high intensities. However, fast metarhodopsin decay and rhodopsin regeneration also play an important role in controlling metarhodopsin in green receptors, resulting in a high rhodopsin content at low light intensities and a reduced overall visual pigment content in bright light. A simple model for the visual pigment–arrestin cycle is used to illustrate the dependence of the visual pigment population states on light intensity, arrestin levels and pigment turnover.     

Correlating species and spectral diversities using hyperspectral remote sensing in early‐successional fields

Advances in remote sensing technology can help estimate biodiversity at large spatial extents. To assess whether we could use hyperspectral visible near‐infrared (VNIR) spectra to estimate species diversity, we examined the correlations between species diversity and spectral diversity in early‐successional abandoned agricultural fields in the Ridge and Valley ecoregion of north‐central Virginia at the Blandy Experimental Farm. We established plant community plots and collected vegetation surveys and ground‐level hyperspectral data from 350 to 1,025 nm wavelengths. We related spectral diversity (standard deviations across spectra) with species diversity (Shannon–Weiner index) and evaluated whether these correlations differed among spectral regions throughout the visible and near‐infrared wavelength regions, and across different spectral transformation techniques. We found positive correlations in the visible regions using band depth data, positive correlations in the near‐infrared region using first derivatives of spectra, and weak to no correlations in the red‐edge region using either of the two spectral transformation techniques. To investigate the role of pigment variability in these correlations, we estimated chlorophyll, carotenoid, and anthocyanin concentrations of five dominant species in the plots using spectral vegetation indices. Although interspecific variability in pigment levels exceeded intraspecific variability, chlorophyll was more varied within species than carotenoids and anthocyanins, contributing to the lack of correlation between species diversity and spectral diversity in the red‐edge region. Interspecific differences in pigment levels, however, made it possible to differentiate these species remotely, contributing to the species‐spectral diversity correlations. VNIR spectra can be used to estimate species diversity, but the relationships depend on the spectral region examined and the spectral transformation technique used.     

Photoelectric conversion by bacteriorhodopsin in charged synthetic membranes.

Photoelectroactivity of oriented purple membrane layers attached to an ion exchange film has been investigated. The action spectrum of the photocurrent followed the absorption spectrum of bacteriorhodopsin. The intactness of structure and function of bacteriorhodopsin was demonstrated by studies of absorption and photocycle kinetics. The direction of the photocurrent suggests that the extracellular surface of purple membrane is more positive. Photocurrents as high as 20 microA cm-2 were obtained in some preparations. The dependence of steady-state photocurrents on intensity of illumination and temperature was also studied. The initial rate of build-up of photocurrent depends linearly on the intensity of illumination while the off rate does not exhibit any dependence on the intensity of illumination. With rise in temperature an increase in the steady state photocurrent has been observed. This dependence was found to be linear when increase of the photocurrent due to proton translocation alone was considered.