PHYSICAL AND ULTRASTRUCTURAL BASIS OF BLUE LEAF IRIDESCENCE IN TWO NEOTROPICAL FERNS

Iridescent blue leaf coloration in two neotropical ferns, Danaea nodosa (L.) Sm. (Marattiaceae) and Trichomanes elegans L. C. Rich. (Hymenophyllaceae), is caused by thin film constructive interference. The ultrastructural basis for the film in D. nodosa is multiple layers of cellulose microfibrils in the adaxial cell walls of the adaxial epidermis. The apparent helicoidal arrangement of the fibrils is analogous to similar color production in arthropods. In T. elegans the blue-green coloration is caused by the remarkably uniform thickness and arrangement of grana in specialized chloroplasts adjacent to the adaxial wall of the adaxial epidermis. The selective advantage of this color production, if any, is unknown but apparently different from that previously studied in Selaginella.     

Blue light inhibits stomatal development in soybean isolines containing kaempferol-3-O-2G-glycosyl-gentiobioside (K9), a unique flavonoid glycoside

Stomata have a fundamental role in controlling plant photosynthesis and transpiration, but very little is known about factors controlling stomatal differentiation and development. Lines of soybean that contain a specific flavonol glycoside, kaempferol‐3‐O‐2‐glycosyl‐gentiobioside (K9), as well as greatly reduced stomatal density, especially on the adaxial epidermis, have been identified. The specific effects of blue light photoreceptors on stomatal development in K9 lines and their isoline pairs containing no K9 were studied. Low irradiances of blue light (7% of total photosynthetically active radiation) added to high irradiances from low‐pressure sodium lamps strongly inhibited stomatal development on the adaxial epidermis of K9 lines, but not in isoline pairs differing putatively in only one gene and lacking K9. Overall, blue light slightly increased stomatal density on the abaxial epidermis in all isolines, demonstrating differential regulation of stomatal development in the upper and lower epidermis. Blue light also caused an increase in leaf area in all isolines, indicating that changes in stomatal density were not the non‐specific result of alterations in leaf area. Morphological studies revealed that the blue light‐induced reduction in stomatal density in K9 lines was due to reduced stomatal initiation as well as aborted or abnormal stomatal development. As the phytochrome photostationary state was kept constant, the results indicate that one or more blue light receptors are involved in the control of stomatal development. This system should be useful for the study of mechanisms controlling stomatal development, even if the photo‐inhibitory response is unique to K9 lines.     

Relationship between stomatal conductance and light intensity in leaves of Zea mays L., derived from experiments using the mesophyll as shade

Attached leaves of Zea mays were illuminated with monochromatic light, with either the upper or the lower epidermis facing the light source. The mesophyll absorbed between 99.5 and 99.6% of the red or blue light used. An inversion of the light direction therefore caused a 200- to 250-fold change in the quantum flux into each epidermis. This variation in quantum flux did not affect stomatal conductance. Stomatal conductance was however correlated with intercellular CO₂ concentration, c_i, and the relationship between stomatal conductance and c_i appeared also to remain the same if changes in c_i were brought about by changes in atmospheric CO₂ concentration instead of light. A close inspection of the data showed that stomata of the upper (adaxial) epidermis exhibited a small increase in conductance (<0.1 cm s^-1) in response to blue light that was superimposed on the dominating response to c_i.     

The Evolutionary Decoupling of Behavioral and Color Cues in a Multicomponent Signal in Two Sceloporus Lizards

The study of multicomponent signals in the context of social systems has generated interesting results demonstrating that complex signals are used in many communication systems. The multicomponent signal in the majority of Sceloporus lizards consists of a color signal (blue abdominal coloration) and the behavioral display of the color signal (fullshow behavior). In a small number of species, males have lost the color signal. We staged outdoor trials between conspecific males in two closely related Sceloporus lizards that differ in the presence of blue abdominal coloration. In the species with the evolutionary loss of blue abdominal coloration (Sceloporus virgatus), fullshow behavior is present but reduced compared with that of the species with male abdominal coloration (Sceloporus undulatus consobrinus). In comparison, the mean rates of other behaviors that do not display abdominal skin (push‐up, head‐bob) did not differ between these species. We also found that S. virgatus males were more likely to show a neutral response following the first fullshow during the 60‐min trial. While, S. u. consobrinus males were more likely to respond to the first fullshow with an aggressive response. Thus, in this case, the color signal and the behavioral signal are evolutionarily decoupled because in S. virgatus the loss of the color signal is not coincident with the loss of the behavioral signal.     

Blue lipophorin from Podisus maculiventris

A chromoprotein responsible for the blue coloration of the hemolymph in the spined soldier bug, Podisus maculiventris (Say), was isolated and identified as lipophorin. With the exception of its blue color the lipoprotein shares similar molecular characteristics with the hemolymph lipophorins of other Hemipterans and insects of several different orders. Its ability to carry a blue chromophore, biliverdin IX γ, adds a new feature to this multifunctional lipoprotein. © 1992 Wiley-Liss, Inc.     

Influence of Social Factors on Seasonal Variations in Plasma Testosterone Levels of Microcebus murinus

Blue tail coloration in hatchling skinks (Eumeces fasciatus and E. laticeps) appears to be an antipredatory adaptation that distracts attention away from the body to the tail. The tail itself serves as a decoy that may be autotomized as a final defense against capture. The effectiveness of intact tails in deflecting attacks from the body was 50% against scarlet kingsnakes in the experimental conditions used. Brightness rather than hue presumably accounts for the higher attack frequency on blue than black tails in this study, but the blue color may have evolved in response to avian predation. Repeated predation without ill effects by several predators allows rejection of the hypothesis that the blue tail is aposematic for the predators tested. The hypothesis that blue tails provide stimuli inhibiting aggression or predation by adult male conspecifics is untenable for E. laticeps because adult males readily eat intact hatchlings. Although this study provides no statistical evidence that blue tail coloration inhibits attack by female E. laticeps on hatchlings, the trend of predation rates on blue- and black-tailed hatchlings is in the direction predicted for inhibition.     

Densification of cyanobacteria from a lake leading to production of β‐cyclocitral and related volatile organic compounds and species change

The purpose of the present study was to demonstrate that the lysis with the blue color formation was caused by densification of the cyanobacteria, and related events of the species change in the cyanobacteria were induced by the resulting volatile organic compounds (VOCs), particularly β‐cyclocitral. In order to obtain a high cell density of cyanobacteria in the laboratory, a concentration technique (graduated cylinder method) using the buoyancy of the gas vesicles was successfully used. The collected scum contained mainly Dolichospermum spp. and Microcystis, and the dispersed cyanobacteria were concentrated in the surface layer after several hours and the concentration ratio became approximately 10. The concentrated cyanobacteria were gradually lysed, while some of the cyanobacteria sank to the bottom, which finally died and disappeared. This method has the additional advantage that it is possible to visualize the entire lysis process. During the concentration process, β‐cyclocitral and its oxidation products together with β‐ionone were significantly detected. Because β‐cyclocitral was easily oxidized to the corresponding carboxylic acid, the pH of the water in the graduated cylinder decreased to approximately 6. Under favorable conditions, lysis with the blue color from phycocyanin could be observed due to the acid stress. Overall, the results of the present study were consistent with the hypothesis that VOCs were produced when the cyanobacteria are highly dense, and that the lysis with the blue color formation occurs due to the higher density.     

葛(豆科)叶的解剖学研究

利用光学显微镜和扫描电镜观察了葛(Pueraria lobata)叶的解剖学特征。结果表明,葛叶片的上、下表皮都只有一层表皮细胞,上表皮比下表皮厚。上、下表皮都有腺毛和非腺毛。气孔主要分布在下表皮,下表皮的气孔密度为(261±17)mm-2,上表皮只有(6±3)mm-2。叶肉由两层栅栏组织细胞和一层海绵组织细胞构成。叶肉细胞中有丰富的叶绿体。在栅栏组织和海绵组织之间有一层平行于叶脉的薄壁细胞。叶脉中含有大量的草酸钙晶体。葛叶的这些形态特征与其喜阳、耐旱的特点相适应。     

Warning Signal Efficacy: Assessing the Effects of Color,Iridescence, and Time of Day in the Field

Warning coloration deters predators from attacking distasteful or toxic prey. Signal features that influence warning color effectiveness are not well understood, and in particular, we know very little about how effective short‐wavelength and iridescent colors are as warning color elements in nature and how warning signal effectiveness changes throughout the day. We tested the effect of these factors on predation risk in nature using specimens of the distasteful pipevine swallowtail butterfly, Battus philenor. B. philenor adults display both iridescent blue and diffusely reflecting orange components in their warning signal. We painted B. philenor wings to create five different model types: all‐black, only‐iridescent‐blue, only‐orange, iridescent‐blue‐and‐orange (intact signal), and matte‐blue‐and‐orange. We placed 25 models in each of 14 replicate field sites for 72 h and checked for attacks three times each day. Model type affected the likelihood of attack; only‐orange models were, the only model attacked significantly less than the all‐black model. Iridescence did not enhance or decrease warning signal effectiveness in our experiment because matte‐blue‐and‐orange models were attacked at the same rate as iridescent‐blue‐and‐orange models. Time of day did not differentially affect model type. Video recordings of attacks revealed that insectivorous birds were responsible. The results of this experiment, when taken with previous work, indicate that the response to blue warning coloration is likely dependent on predator experience and context, but that iridescence per se does not affect warning signals in a natural context.     

What's in a band? The function of the color and banding pattern of the Banded Swallowtail

Butterflies have evolved a diversity of color patterns, but the ecological functions for most of these patterns are still poorly understood. The Banded Swallowtail butterfly, Papilio demolion demolion, is a mostly black butterfly with a greenish‐blue band that traverses the wings. The function of this wing pattern remains unknown. Here, we examined the morphology of black and green‐blue colored scales, and how the color and banding pattern affects predation risk in the wild. The protective benefits of the transversal band and of its green‐blue color were tested via the use of paper model replicas of the Banded Swallowtail with variations in band shape and band color in a full factorial design. A variant model where the continuous transversal green‐blue band was shifted and made discontinuous tested the protective benefit of the transversal band, while grayscale variants of the wildtype and distorted band models assessed the protective benefit of the green‐blue color. Paper models of the variants and the wildtype were placed simultaneously in the field with live baits. Wildtype models were the least preyed upon compared with all other variants, while gray models with distorted bands suffered the greatest predation. The color and the continuous band of the Banded Swallowtail hence confer antipredator qualities. We propose that the shape of the band hinders detection of the butterfly's true shape through coincident disruptive coloration; while the green color of the band prevents detection of the butterfly from its background via differential blending. Differential blending is aided by the green‐blue color being due to pigments rather than via structural coloration. Both green and black scales have identical structures, and the scales follow the Bauplan of pigmented scales documented in other Papilio butterflies.     

Plumage color as a status signal in male–male interaction in the red-flanked bushrobin, Tarsiger cyanurus

Recent studies have suggested that structural-based coloration is an honest signal of male genetic and/or conditional quality in sexual selection. However, whether structural coloration functions in intrasexual competition is unknown. We examined whether plumage color functions as a status signal during intrasexual interactions in the red-flanked bushrobin Tarsiger cyanurus; adult males have many blue plumes as structural coloration whereas yearling males and females are olive brown with few blue plumages. Blue males did not always dominate olive-brown males. The number of interactions did not differ with the colors of the two birds involved. The interactions of a blue male and an olive-brown male were less aggressive than those of two blue or of two olive-brown males. In this study, we found that structural plumage coloration may serve as a signal of aggressive intent and lower the escalation level of an aggressive interaction in a manner consistent with hypotheses regarding the evolution of delayed plumage maturation.     

A vacuolar iron transporter in tulip, TgVit1, is responsible for blue coloration in petal cells through iron accumulation

Blue color in flowers is due mainly to anthocyanins, and a considerable part of blue coloration can be attributed to metal-complexed anthocyanins. However, the mechanism of metal ion transport into vacuoles and subsequent flower color development has yet to be fully explored. Previously, we studied the mechanism of blue color development specifically at the bottom of the inner perianth in purple tulip petals of Tulipa gesneriana cv. Murasakizuisho. We found that differences in iron content were associated with the development of blue- and purple-colored cells. Here, we identify a vacuolar iron transporter in T. gesneriana ( TgVit1 ), and characterize the localization and function of this transporter protein in tulip petals. The amino acid sequence of TgVit1 is 85% similar that of the Arabidopsis thaliana vacuolar iron transporter AtVIT1, and also showed similarity to the AtVIT1 homolog in yeast, Ca²⁺-sensitive cross-complementer 1 (CCC1). The gene TgVit1 was expressed exclusively in blue-colored epidermal cells, and protein levels increased with increasing mRNA expression and blue coloration. Transient expression experiments revealed that TgVit1 localizes to the vacuolar membrane, and is responsible for the development of the blue color in purple cells. Expression of TgVit1 in yeast rescued the growth defect of ccc1 mutant cells in the presence of high concentrations of FeSO₄. Our results indicate that TgVit1 plays an essential role in blue coloration as a vacuolar iron transporter in tulip petals. These results suggest a new role for involvement of a vacuolar iron transporter in blue flower color development.     

Attractive blue‐green egg coloration and cuckoo−host coevolution

Recent evidence suggests that blue‐green coloration of bird eggshells may be related to female and/or egg phenotypic quality, and that such colour may affect parental effort and therefore the nutritional environment of developing nestlings. Here we suggest that these relationships and the signal function of eggshell coloration would affect the outcome of coevolution between avian brood parasites and their hosts in at least three different non‐exclusive evolutionary pathways. First, by laying blue‐green coloured eggs, cuckoo females may exploit possible sensory biases of their hosts, constraining the evolution of parasitic egg recognition, and thus avoid rejection. Second, because of the relatively high costs of laying blue eggs, cuckoo females may be limited in their ability to mimic costly blue‐green eggs of their hosts because cuckoo females lay many more eggs than their hosts. Furthermore, costs associated with foreign egg recognition errors would be relatively higher for hosts laying blue eggs. Third, cuckoos may use coloration of host eggs for selecting individuals or specific hosts of appropriate phenotypic quality (i.e. parental abilities). We here explored some predictions emerging from the above scenarios and found partial support for two of them by studying egg coloration of European cuckoos (Cuculus canorus) and that of their 25 main hosts, as well as parasitism and rejection rate of hosts. Cuckoo hosts parasitized with more blue, green, and ultraviolet cuckoo eggs, or those laying more blue‐green eggs, were more prone to accept experimental parasitism with artificial cuckoo eggs. In addition, coloration of cuckoo eggs is more variable when parasitizing hosts laying bluer‐greener eggs, even after controlling for the effect of host egg coloration (i.e. degree of egg matching). Globally, our results are consistent with the proposed hypothesis that host egg traits that are related to phenotypic quality of hosts, such as egg coloration, may have important implications for the coevolutionary interaction between hosts and brood parasites. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106 , 154–168.     

Species‐dependent microarchitectural traits of iridescent scales in the triad taxa of Ornithoptera birdwing butterflies

Ornithoptera birdwing butterflies have blue, green, or orange iridescent scales in different species or subspecies. To understand the species‐ or subspecies‐dependent scale color differences, we performed comparative morphometric analyses of iridescent scales from three closely related taxa: O. priamus priamus (green), O. priamus urvillianus (blue), and O. croesus (orange). The three types of Ornithoptera wings exhibited reversible color changes to longer wavelengths with different kinetics upon immersion in methanol, suggesting that their color differences are at least partly based on differences in the size of air cavities made by nanostructures. Cover scales of all three color types were visually semi‐transparent glass scales that exhibited color when placed on a dark background. The dorsoventral differences in coloration were observed in single scales, suggesting the optical importance of scale surfaces. Scanning electron microscopy of cover scales in cross section revealed that all color types exhibited finely sculpted tapered ridges and thick, irregular basal multilayers containing tandemly clustered granular objects and air cavities. Scale thickness, ridge height, and multilayer thickness were significantly different among the three color types, and granular object size was significantly different between orange scales and blue and green scales. We conclude that each of the three taxa of Ornithoptera butterflies possesses unique quantitative size values on tapered ridges and irregular multilayers with granular objects and air cavities to express unique structural color. These species‐ or subspecies‐dependent structural colors might have evolved via quantitative shifts in these microarchitectural traits rather than via changes in the basic developmental or architectural plan for color expression.     

The evolution of black plumage from blue in Australian fairy‐wrens (Maluridae): genetic and structural evidence

Genetic variation in the melanocortin‐1 receptor (MC1R) locus is responsible for color variation, particularly melanism, in many groups of vertebrates. Fairy‐wrens, Maluridae, are a family of Australian and New Guinean passerines with several instances of dramatic shifts in plumage coloration, both intra‐ and inter‐specifically. A number of these color changes are from bright blue to black plumage. In this study, we examined sequence variation at the MC1R locus in most genera and species of fairy‐wrens. Our primary focus was subspecies of the white‐winged fairy‐wren Malurus leucopterus in which two subspecies, each endemic to islands off the western Australian coast, are black while the mainland subspecies is blue. We found fourteen variable amino acid residues within M. leucopterus, but at only one position were alleles perfectly correlated with plumage color. Comparison with other fairy‐wren species showed that the blue mainland subspecies, not the black island subspecies, had a unique genotype. Examination of MC1R protein sequence variation across our sample of fairy‐wrens revealed no correlation between plumage color and sequence in this group. We thus conclude that amino acid changes in the MC1R locus are not directly responsible for the black plumage of the island subspecies of M. leucopterus. Our examination of the nanostructure of feathers from both black and blue subspecies of M. leucopterus and other black and blue fairy‐wren species clarifies the evolution of black plumage in this family. Our data indicate that the black white‐winged fairy‐wrens evolved from blue ancestors because vestiges of the nanostructure required for the production of blue coloration exist within their black feathers. Based on our phylogeographic analysis of M. leucopterus, in which the two black subspecies do not appear to be each other's closest relatives, we infer that there have been two independent evolutionary transitions from blue to black plumage. A third potential transition from blue to black appears to have occurred in a sister clade.     

Abaxial and adaxial stomata from Pima cotton (Gossypium barbadense L.) differ in their pigment content and sensitivity to light quality

Sensitivity to light quality and pigment composition were analysed and compared in abaxial and adaxial stomata of Gossypium barbadense L. (Pima cotton). In most plants, abaxial (lower) stomatal conductances are higher than adaxial (upper) ones, and stomatal opening is more sensitive to blue light than to red. In greenhouse-grown Pima cotton, abaxial stomatal conductances were two to three times higher than adaxial ones. In contrast, adaxial stomatal conductances were 1·5 to two times higher than abaxial ones in leaves from growth chamber-grown plants. To establish whether light quality was a factor in the regulation of the relationship between abaxial and adaxial stomatal conductances, growth-chamber-grown plants were exposed to solar radiation outdoors and to increased red light in the growth chamber. In both cases, the ratios of adaxial to abaxial stomatal conductance reverted to those typical of greenhouse plants. We investigated the hypothesis that adaxial stomata are more sensitive to blue light and abaxial stomata are more sensitive to red light. Measurements of stomatal apertures in mechanically isolated epidermal peels from growth chamber and greenhouse plants showed that adaxial stomata opened more under blue light than under red light, while abaxial stomata had the opposite response. Using HPLC, we quantified the chlorophylls and carotenoids extracted from isolated adaxial and abaxial guard cells. All pigments analysed were more abundant in the adaxial than in the abaxial guard cells. Antheraxanthin and β-carotene contents were 2·3 times higher in adaxial than in abaxial guard cells, comparing with ad/ab ratios of 1·5–1·9 for the other pigments. We conclude that adaxial and abaxial stomata from Pima cotton have a differential sensitivity to light quality and their distinct responses are correlated with different pigment content.     

No Evidence that Male Choice Contributes to the Maintenance of a Shared, Sex-Limited Trait in Mimetic and Non-mimetic Female Tiger Swallowtail Butterflies, Papilio glaucus

In animal species that have morphological polymorphisms maintained by unique or divergent selection pressures, understanding the preservation of shared traits is important for identifying the factors that are influencing overall evolutionary processes. In the Eastern tiger swallowtail butterfly, Papilio glaucus, females are dimorphic. One morph (‘dark-morph’) is mostly black and mimics the toxic pipevine swallowtail, Battus philenor. These females have large amounts of blue coloration on the dorsal hind wings that enhances their mimetic resemblance. Conversely, the alternate female type (‘yellow-morph’) is similar to males in coloration with the exception of extensive dorsal blue coloration, comparable to dark-morph females. Such coloration is almost completely absent in males. We examined dorsal blue coloration in P. glaucus to determine if mimetic resemblance in dark morphs is predominantly responsible for the maintenance of dorsal blue color in both female types, or whether mate recognition and/or sexual selection by males has a stronger influence on this trait. We measured the relative amount and variance of dorsal and ventral blue coloration in females of both color morphs, as well as males. We also compared these measurements to similar ones taken in the sister species, P. canadensis (which does not exhibit female dimorphism). Lastly, we investigated mate recognition and preferences of wild males. Our results suggest that mimetic resemblance may be more important than sexual selection for sustaining dorsal blue coloration in dark-morph females and that yellow-morphs could have elevated levels of blue due to currently unknown genetic associations. Although trait correlation between sexes is common, intrasexual trait correlation in a sex-limited, polymorphic species has not been frequently observed.     

Inhibition of choline oxidase by quinoid dyes

Choline oxidase catalyzes the oxidation of choline to glycine-betaine, with betaine-aldehyde as intermediate and molecular oxygen as primary electron acceptor. This study reports on the inhibitory effects of triarylmethanes (cationic malachite green; neutral leukomalachite green), phenoxazines (cationic, meldola blue and nile blue; neutral nile red) and a structurally-related phenothiazine (methylene blue) on choline oxidase, assayed at 25°C in 50 mM MOPS buffer, pH 7, using choline as substrate. Methylene B acted as a competitive inhibitor with K_i = 74 ± 7.2 μM, pointing to the choline–binding site of the enzyme as a target site. Nile B caused noncompetitive inhibition of enzyme activity with K_i = 20 ± 4.5 μM. In contrast to methylene B and nile B, malachite G and meldola B caused complex, nonlinear inhibition of choline oxidase, with estimated K_i values in the micromolar range. The difference in kinetic pattern was ascribed to the differential ability of the dyes to interact (and interfere) with the flavin cofactor, generating different perturbations in the steady-state balance of the catalytic process.     

Guard cells on adaxial and abaxial epidermes of <Emphasis Type="Italic">Erythrina corallodendron</Emphasis> sepals

This study investigated guard cells on the adaxial and the abaxial epidermes during Erythrina corallodendron sepal development. On the adaxial epidermis, the morphology of guard cells was highly variable and changes in aperture induced by abscisic acid (ABA) were observed in 9.1 % stomata, while on the abaxial epidermis 86.7 % stomata responded to ABA. On the adaxial epidermis, stomata did not close even when guard cell pressure potential was reduced to zero by plasmolysis, even if fluorescein diacetate revealed that guard cells were alive. It was concluded that guard cells on the adaxial and the abaxial epidermes of sepals sensed environmental conditions differently, maybe due to different guard cell wall elasticity.