Screening Pigment Research Articles

A critical assessment of the analysis and distributions of scytonemin and related UV screening pigments in sediments

High performance liquid chromatography-mass spectrometry has been used to identify the cyanophyta-derived ultraviolet screening pigment scytonemin and its reduced counterpart in an Antarctic lake sediment. The formation of an artefact during the analysis has been demonstrated, enabling the recovery of improved estimates of the scytonemin and reduced scytonemin signals. The overall pigment composition records a primary producer community comprising both oxygenic and anoxygenic photoautotrophs, with variations in the abundance of scytonemin relative to the chlorophylls and carotenoids that are consistent with changes in response to variations in the light regime over time.

Sequential reduction of UV-B radiation in the field alters the pigmentation of an Antarctic leafy liverwort

UV-B radiation (280–315 nm), incident on the leafy liverwort Cephaloziella varians growing at Rothera Point on the Western Antarctic Peninsula, was manipulated in late austral spring 1998 by screens consisting of a novel combination of Plexiglas panes and polyester sheets. The screens transmitted approximately 79, 68, 48, 41 and 31% of daily UV-B radiation dose. Plants from under and outside the screens were sampled seven times over 35 days. Correlative analyses indicated that concentrations of an anthocyanin-like pigment located in leaf tips were positively associated with UV-B transmission at the last two samplings, that concentrations of total chlorophylls were negatively associated with UV-B transmission at the final sampling, and that those of UV-B screening pigments were positively correlated with transmission at three intermediate samplings. Plants exposed to low UV-B transmission levels were visibly greener than those exposed to high transmission levels after approximately 23 days. The closest relationships between the treatment and concentrations of UV-B screening pigments were associated with increased ambient biologically weighted UV-B dose received by plants in the 5.5 h before each sampling, which coincided with the passage of the ozone hole over Rothera Point. As C. varians emerged from melting snow and ice, concentrations of the anthocyanin-like pigment and chlorophyll respectively increased and decreased faster in plants exposed to high transmission levels of UV-B, relative to those exposed to low UV-B transmission levels.

Vision and visual variation in the peacock blenny

Microspectrophotometry (MSP) revealed the presence of a rod and at least two cone classes (mid‐ and long‐wavelength sensitive) in the intertidal peacock blenny Salaria pavo. Both rhodopsin and porphyropsin based visual pigments were found in all fish, together with high individual variation in the chromophore ratio. The three morphs (females, males and sneaker males) differed in their spectral sensitivities (as measured with the optomotor response) with sneakers having higher sensitivity at long‐wavelengths than either males or females. This long‐wave displacement of peak sensitivity could be due to elevated proportions of porphyropsin visual pigments in the sneakers’ retinae. The lenses of all morphs exhibited a short‐wavelength cut‐off and an unusual layer of carotenoid was found behind the retinal pigment epithelium and in the outer segments of some cones. These screening pigments could serve a photo‐protective role or to improve visual contrast. No short‐wave photoreceptors were located using MSP. This indicates that this cone class may be absent or present at very low numbers in the retina. This is the first in depth study of the visual system of a blenniid fish and indicates potential within‐species visual variation that may be related to the species’ habitat and morph‐specific behavioural requirements.

Effects of UV irradiation on barley and tomato leaves: thermoluminescence as a method to screen the impact of UV radiation on crop plants

The effect of different UV intensities and irradiation times on barley and tomato leaves was investigated by analysis of thermoluminescence (TL) and chlorophyll (chl) fluorescence measurements. Epifluorescence microscopy was used to estimate the epidermal UV transmittance of leaves. In barley a strong supression of TL emission from the S2QB- (B-band) and the S2QA- (Q-band) charge recombination was observed increasing with prolonged UV exposure. Primary barley leaves were more sensitive to UV than secondary leaves. In tomato plants a decrease in the B-band only takes place at very high UV intensities and after prolonged exposure times (4 h). The impact of UV in cotyledons was more pronounced than in pinnate leaves of tomato plants. The strong differences in sensitivity to UV in the investigated barley and tomato variety may be due to different concentrations of UV screening pigments in the epidermal layer as demonstrated by epifluorescence measurements. The results show that TL has the same potential to analyse the sensitivity or tolerance of crop plants to UV irradiation as routine fluorescence techniques. Furthermore, TL is directly monitoring the radical pair states of PSII and can distinguish between UV-induced donor and acceptor site-related damage.

Coexpression of Two Visual Pigments in a Photoreceptor Causes an Abnormally Broad Spectral Sensitivity in the Eye of the ButterflyPapilio xuthus

The compound eye of the butterfly Papilio xuthus consists of three different types of ommatidia, each containing nine photoreceptor cells (R1-R9). We have found previously that the R5-R8 photoreceptors of type II ommatidia coexpress two different mRNAs, encoding opsins of green- and orange-red-absorbing visual pigments (Kitamoto et al., 1998). Do these cells contain two functionally distinct visual pigments? First, we identified the sensitivity spectrum of these photoreceptors by using combined intracellular recording and dye injection. We thus found that the R5-R8 of type II ommatidia have a characteristic sensitivity spectrum extending over an excessively broad spectral range, from the violet to the red region; the photoreceptors are therefore termed broadband photoreceptors. The spectral shape was interpreted with a computational model for type II ommatidia, containing a UV visual pigment in cells R1 and R2, two green visual pigments in cells R3 and R4, a far-UV-absorbing screening pigment (3-hydroxyretinol) in the distal part of the ommatidium, and a red-screening pigment that surrounds the rhabdom. The modeling suggests that both visual pigments in the R5-R8 photoreceptors participate in phototransduction. This work provides the first compelling evidence that multiple visual pigments participate in phototransduction in single invertebrate photoreceptors.

48 The effects of environmental factors on the resumption of photosynthetic activity of cyanobacterial mats following rehydration

Extensive cyanobacterial mats cover the intertidal zone near Guerrero Negro, Baja California Sur. These mats are exposed to extreme desiccation and osmotic stress between tidal flows and rains, and spend most of the time dry and metabolically inactive. Therefore, periods of hydration are extremely important for growth as well as for repair of cellular damage from desiccation and ultraviolet radiation (UVR) accrued when the mat is dry. PAM fluorometry in conjunction with carbon incorporation assays were used to determine the effects of salinity, irradiance and UVR on the recovery of photosynthetic activity in these mats after an extended period of desiccation. The mat used in our study was primary composed of Lyngbya sp. Photosynthetic activity recovery rates (using PAM fluorometry) decreased with increasing salinity. This trend was similar under high and low light intensities, but rates were significantly lower under low light. Alternatively, the carbon incorporation method showed rates increased faster in salinities of 27 and 55 ppt than in salinities of 0 or 75 ppt. The Lyngbya mat also failed to recover photosynthetic potential in the dark. Although these mats recovered faster under high intensity light, the effect of salinity on photosynthesis is more complex. UVR did not affect the recovery of photosynthetic activity, no matter which method was used. This lack of effect is most likely due to the high content of the UVR screening pigment, scytonemin, in the upper layer of the mat.

Twilight orientation to polarised light in the crepuscular dung beetle Scarabaeus zambesianus.

The polarisation pattern of skylight offers many arthropods a reference for visual compass orientation. The dung beetle Scarabaeus zambesianus starts foraging at around sunset. After locating a source of fresh droppings, it forms a ball of dung and rolls it off at high speed to escape competition at and around the dung pile. Using behavioural experiments in the field and in the laboratory, we show that the beetle is able to roll along a straight path by using the polarised light pattern of evening skylight. The receptors used to detect this skylight cue can be found in the ommatidia of the dorsal rim area of the eye, whose structures differ from the regular ommatidia in the rest of the eye. The dorsal rim ommatidia are characterised by rhabdoms with microvilli oriented at only two orthogonal orientations. Together with the finding that the receptors do not twist along the length of the rhabdom, this indicates that the photoreceptors of the dorsal rim area are polarisation sensitive. Large rhabdoms, a reflecting tracheal sheath and a lack of screening pigments make this area of the eye well adapted for polarised light detection at low light levels. The fan-shaped arrangement of receptors over the dorsal rim area was previously believed to be an adaptation to polarised light analysis, but here we argue that it is simply a consequence of the way that the eye is built.

Longitudinal leaf gradients of UV-absorbing screening pigments in barley (Hordeum vulgare).

In vivo leaf characteristics were examined to describe longitudinal gradients of UV-absorbing screening pigments in barley. Chlorophyll fluorescence properties and in vivo absorption spectra (210-750 nm) of leaves were measured from the base to the tip. Barley leaves showed strong longitudinal gradients of chlorophyll, where chlorophyll concentration increased within the first 5-8 cm from the leaf base, and did not significantly change for the remaining part of the leaf. Fluorescence microscopy was used to localize cell wall bound screening pigments different from flavonoids, since flavonoids lack a blue-green fluorescence emission (Lichtenthaler and Schweiger 1998). Measurements of in vivo chlorophyll fluorescence indicated that the ratio of UV-absorbing screening pigments per leaf area increases from the leaf base to the tip. These gradients were confirmed by in vivo absorption spectra. It is demonstrated that leaves in the early stage of development are less protected against UV-radiation than fully developed mature leaf regions. The experiments show that measurements of in vivo chlorophyll fluorescence are ideally suited as a fast non-invasive tool to estimate the epidermal UV-transmittance in different leaf sections.

UV-B radiation arising from stratospheric ozone depletion influences the pigmentation of the Antarctic moss Andreaea regularis.

Changes to the radiative environment arising from stratospheric ozone (O(3)) depletion and subsequent associations between these changes and the pigmentation of the moss Andreaea regularis were measured in late austral spring and early summer 1998 at Rothera Point on the western Antarctic Peninsula (67 degrees S, 68 degrees W). A strong relationship between O(3) column depth and the ratio of UV-B to PAR irradiance ( F(uv-b)/ F(par)) was recorded at ground level ( r(2)=92%, P<0.001). Weaker, but significant, associations between O(3) column depth and ground level unweighted and biologically effective UV-B radiation (UV-B(be)) were also found. Regression analyses indicated that F(uv-b)/ F(par) was the best predictor for concentrations of UV-B screening pigments and total carotenoids extracted from plant tissues. Concentrations of these pigments were loosely ( r(2)= ca. 30%) but significantly ( P<0.01) positively associated with F(uv-b)/ F(par). Concentrations of UV-B screening pigments were also positively associated with irradiances and daily doses of unweighted UV-B and UV-B(be) radiation. The concentrations of chlorophylls a and b were apparently unaffected by O(3) depletion. The data derived from this study suggest that changes to the radiative environment associated with stratospheric O(3) depletion influence the pigmentation of A. regularis. As a corollary, flavonoids are shown to be present in tissues of A. regularis.

The Nymphal Eyes of Parabuthus transvaalicus Purcell, 1899 (Buthidae): An Accessory Lateral Eye in a Scorpion

In P. transvaalicus nymphs, 5 pairs of lateral ocelli each composed of a corneal lens, R-cell units forming a latticed rhabdom, arhabdomeric cells and pigment cells are present. In addition, we found a pair of unpigmented accessory sense organs situated ventroposteriorly to the lateral ocelli in prenymphs as well as in first nymphs. They are composed of primary, rhabdomeric sensory cells, and we infer that they represent a second type of lateral eye. They also comprise sensory units, but lenses and screening pigment are lacking. Their position and cellular architecture corresponds well with that of the “rudimentary” lateral eye of the xiphosuran, Limulus. The occurrence of a bipartite lateral visual system in Chelicerata and Arthropoda is discussed.

Textile Ink Jet Printing with Low Viscosity Pigment Inks

The development of textile pigment ink solutions is a very complex area were currently available systems have been tailored to individual ink jet print head technologies.The present paper will outline developments in the area of novel polymer dispersing agents for pigment inks, which allows the production of textile pigment inks with low viscosity profiles suitable for piezo print heads.These new textile pigment ink jet inks produce ink jet prints, which are comparable to traditional textile pigment screen prints.

The fine structure of the ocelli of Triatoma infestans (Hemiptera: Reduviidae)

The morphology and fine structure of the ocelli of Triatoma infestans have been analyzed by means of light and electron microscopy. The two dorsal ocelli of this species are located behind the compound eyes, looking dorsally and frontally. Externally, the ocelli are marked by the corneal lenses virtually spherical in form and limited internally by a cuticular apodeme. The lens focuses the incoming rays beyond the retina. A single layer of corneagen cells lies below the cuticular lens. The corneagen cells and photoreceptors are arranged in a cup-like fashion beneath the cuticular lens. A distal retinal zone comprises the rhabdoms, which are laterally connected in an hexagonal meshwork. A middle retinal zone comprises the receptor cell segment free of rhabdom, and a proximal zone their axons. In the middle zone, the oviform nuclei and spheroids are located. Screening pigment granules are present within the retinal cell. Spherical mitochondria are homogeneously distributed in the cytoplasm of the cell body. In the axonal zone, mitochondria are found in the peripheral region. Axons from receptor cells extend into the ocellar neuropile at the base of the ocelli, to synapse with second order neurons. The large axons of second order neurons are bundled by glial cells. The ocellar plexus exhibits a high diversity of synaptic unions (i.e. axo-dendritic, axo-axonic, dendro-axonic, and dendro-dendritic).

Photopigment gene expression and rhabdom formation in the crayfish (Procambarus clarkii).

This study examines the expression of the photopigment gene in the developing retina of the freshwater crayfish Procambarus clarkii(Crustacea, Malacostraca, Decapoda). Both sense and anti-sense RNA probes were used for in situ hybridization (ISH) of whole embryos collected at various stages during development. A characteristic of retinal development is the formation of screening pigment in the retinular cells of the retinal ommatidia. This pigmentation is seen as a band that begins at the lateral side of the retinal field and progresses medially. At hatching the retina is approximately 50% pigmented. ISH of whole embryos shows that expression of the photopigment gene by the retinular cells correlates with the extent of the screening pigment band in the retina and with the presence of rhabdoms within the ommatidia. Sections taken through embryos after being hybridized indicate that staining is localized in the cytoplasm of the retinular cells and in the axonal region below the basement membrane. No staining reaction was seen in the rhabdoms of older ommatidia. ISH staining was also seen at the anterior midline of the protocerebrum where extraretinal photoreceptors have been reported. The data presented here show a close correlation of opsin expression within the retinular cells of the ommatidia and the formation of the very early rhabdoms, similar to Drosophila. The results will be discussed in relation to recent studies in Drosophila that suggest rhodopsin plays a role in effecting the organization of the terminal web-like cytoskeleton at the base of the developing rhabdom microvilli.

Response of two Antarctic bryophytes to stratospheric ozone depletion

AbstractWe report a study which measured changes to the radiative environment arising from stratospheric O3 depletion at Rothera Point on the western Antarctic Peninsula (67°S, 68°W) and subsequent associations between these changes and the pigmentation and maximum quantum yield of photochemistry (Fv/Fm) of two Antarctic bryophytes, the liverwort Cephaloziella varians and the moss Sanionia uncinata. We found a strong relationship between O3 column depth and the ratio of UV‐B to PAR irradiance (FUV‐B/FPAR) recorded at ground level. Weaker, but significant, associations were also found between O3 column depth and noon irradiances and daily doses of unweighted and biologically effective UV‐B radiation received at ground level. Regression analyses indicated that FUV‐B/FPAR and daily dose of unweighted UV‐B were best predictors for concentrations of total carotenoids and UV‐B screening pigments extracted from bryophyte tissues. Concentrations of these pigments were loosely but significantly positively associated with O3‐dependent irradiance parameters. HPLC analyses of carotenoids also suggested that both species increased the synthesis of neoxanthin during periods of O3 depletion. Violaxanthin, lutein, zeaxanthin and b,bββ‐carotene concentrations were also apparently influenced by O3 reduction, but not consistently across both bryophyte species. Concentrations of chlorophylls a and b were apparently unaffected by O3 depletion.No direct associations between Fv/Fm and O3‐dependent irradiance parameters were found. However stepwise multiple regression analyses suggested that the production of UV‐B screening pigments conferred protection from elevated FUV‐B/FPAR on Fv/Fm in both species and that carotenoids conferred protection on Fv/Fm in Sanionia. Our data suggest that changes to the radiative environment associated with stratospheric O3 depletion influence the pigmentation of two Antarctic bryophytes, but that Fv/Fm is unaffected, at least in part because of rapid synthesis of protective pigments.

Fine structure of the lateral eyes in the tiny marine Antarctic mite Halacarellus thomasi : an ultrastructural study

The lateral eyes of the tiny marine /-Vntarclic mite, Halacarellus iliomasi, consist of two entities in the same eye capsule. The oplical axes of the two eyes point in different directions and the two eyes are of different sizes- The larger of the two is covered by a circular lens of approximately 13 |im diameter, with considerably greater inner than outer convex curvature. The cover of the smaller eye consists of little else than a circular 10-).m'i-\vide transparent patch of cuticle. Both eyes have an everted retina with few (4-6) retinula cells, and their microvilli, which are oriented perpendicular to the light' path, have diameters of 55 nm. Screening pigments in the form of lipid-like, electron-opaque cell inclusions, measuring 1-2 um in size, are abundant on the proximal side of the retina. A small l-)im-thick muscle is in con­ tact with the eye capsule. On the basis of our structural data and comparisons with other mite species, we con­ clude that form, colour and polarization vision would play no role in the eye of /-/, ihouiasi. More likely, (he function of the eye is to monitor seasonal daylength changes to allow synchronization of reproductive activities.

Evidence for a two pigment visual system in the fiddler crab, Uca thayeri.

Intraocular recordings were made from the eyestalks of dark-adapted fiddler crabs (Uca thayeri) during presentation of monochromatic light flashes of different wavelengths and intensities. Two types of signals were recorded in different experiments: slow potentials (electroretinogram) and fast potentials (spikes). The latter were also recorded in the presence of a continuous green or red adapting light. The resulting visual spectral-sensitivity curves, when fitted to rhodopsin-based visual pigment absorption spectra (from Dartnall nomograms), indicated the presence of two visual pigments, one with an absorption maximum near 430 nm, and the other with a peak absorption between 500 nm and 540 nm. The data also provided evidence for some differential bleaching of the pigments in the presence of a colored adapting light, but most of the adaptation effect was probably due to changes in screening pigment and neural desensitization or inhibition. These two observations suggest that an adequate substrate for color vision may exist in this and other species of fiddler crabs. The electroretinogram and spike-recording methods produced similar visual-sensitivity data, suggesting that latter technique, a much more efficient way of collecting data that is physiologically relevant, may be the method of choice for determining spectral sensitivity in crustaceans.

Colour in the eyes of insects.

Many insect species have darkly coloured eyes, but distinct colours or patterns are frequently featured. A number of exemplary cases of flies and butterflies are discussed to illustrate our present knowledge of the physical basis of eye colours, their functional background, and the implications for insect colour vision. The screening pigments in the pigment cells commonly determine the eye colour. The red screening pigments of fly eyes and the dorsal eye regions of dragonflies allow stray light to photochemically restore photoconverted visual pigments. A similar role is played by yellow pigment granules inside the photoreceptor cells which function as a light-controlling pupil. Most insect eyes contain black screening pigments which prevent stray light to produce background noise in the photoreceptors. The eyes of tabanid flies are marked by strong metallic colours, due to multilayers in the corneal facet lenses. The corneal multilayers in the gold-green eyes of the deer fly Chrysops relictus reduce the lens transmission in the orange-green, thus narrowing the sensitivity spectrum of photoreceptors having a green absorbing rhodopsin. The tapetum in the eyes of butterflies probably enhances the spectral sensitivity of proximal long-wavelength photoreceptors. Pigment granules lining the rhabdom fine-tune the sensitivity spectra.

Spectrophotometric assessment of leaf UV-B absorbing compounds and chemically determined total phenolic levels are strongly correlated

Investigations concerning the two major ecological roles of phenolics use different methods to quantify these compounds (phenolics as antiherbivores, Folin-Ciocalteu chemical assay; phenolics as UV-B screening pigments, UV-B absorbance). Yet, comparisons of the corresponding results are not possible, since an empirical correlation between the two methods is lacking. In the present study, significant regressions between total phenolic levels (chemically determined with the Folin-Ciocalteu method) and leaf UV-B absorbing capacity (assessed from simple absorbance measurements of methanolic extracts at 300 nm) were found in all seven plant species tested, yet interspecies differences in regression equations were evident. Provided that a standard curve between UV-absorbance versus total phenolic levels is established for each test plant, the latter could be predicted from the former. The UV-absorbance method is preferable because it is time-saving, simpler, and less costly. Given the strong regression between the two variables, a comparison of the generalizations reached by the two lines of research using the corresponding methods for phenolic determination is attempted.Key words: UV-B absorbing capacity, phenolics, herbivory, Mediterranean plants.

Ultrastructure and orientation of ommatidia in the dorsal rim area of the locust compound eye

In many insect species, a dorsal rim area (DRA) in the compound eye is adapted to analyze the sky polarization pattern for compass orientation. In the desert locust Schistocerca gregaria, these specializations are particularly striking. The DRA of the locust consists of about 400 ommatidia. The facets have an irregular shape, and pore canals are often present in the corneae. Screening pigment is missing in the region of the dioptric apparatus suggesting large receptive fields. The rhabdoms are shorter, but about four times larger in cross-section than the rhabdoms of ordinary ommatida. Eight retinula cells contribute to the rhabdom. The microvilli of retinula cell 7 and of cells 1, 2, 5, 6, 8 are highly aligned throughout the rhabdom and form two blocks of orthogonal orientation. The microvilli in the minute rhabdomeres of retinula cells 3 and 4, in contrast, show no particular alignment. As in other insect species, microvillar orientations are arranged in a fan-like pattern across the DRA. Photoreceptor axons project to distinct areas in the dorsal lamina and medulla. The morphological specializations in the DRA of the locust eye most likely maximize the polarization sensitivity and suggest that the locust uses this eye region for analysis of the sky polarization pattern.

Reflections on colourful ommatidia of butterfly eyes.

The eye shine of butterflies from a large number of ommatidia was observed with a modified epi-illumination apparatus equipped with an objective lens of large numerical aperture. A few representative cases are presented: the satyrine Bicyclus anynana, the heliconian Heliconius melpomene, the small white Pieris rapae and the small copper Lycaena phlaeas. The colour of the eye shine is determined mainly by the reflectance spectrum of the tapetal mirror and the transmittance spectrum of the photoreceptor screening pigments, if present near the light-guiding rhabdom. Reflectance spectra measured from individual ommatidia show that tapetum and screening pigments are co-expressed in fixed combinations, thus determining different ommatidial classes. The classes are distributed in an irregular pattern that can be rapidly assessed with the novel epi-illumination apparatus. Many butterfly species appear to have red-reflecting ommatidia, which is interpreted to indicate the presence of red-sensitive photoreceptors.