Caution with colour calculations: spectral purity is a poor descriptor of flower colour visibility.

BACKGROUND: The colours of flowers are of key interest to plant and pollination biologists. An increasing number of studies have investigated the importance of saturation of flower colours (often called 'spectral purity' or 'chroma') for visibility to pollinators, but the conceptual, physiological and behavioural foundations for these metrics as well as the calculations used rest on slender foundations. METHODS: We discuss the caveats of colour attributes that are derived from human perception, and in particular spectral purity and chroma, as variables in flower colour analysis. We re-analysed seven published datasets encompassing 774 measured reflectance spectra to test for correlations between colour contrast, spectral purity and chroma. MAIN FINDINGS AND CONCLUSIONS: We identify several concerns with common calculation procedures in animal colour spaces. Studies on animal colour vision provide no ground to assume that any pollinator perceives (or responds to) saturation, chroma or spectral purity in the way humans do. A re-analysis of published datasets revealed that values for colour contrast between flowers and their background are highly correlated with measures for spectral purity and chroma, which invalidates treating these factors as independent variables as is currently commonplace. Strikingly, spectral purity and chroma - both of which are metrics for saturation and are often used synonymously - are not correlated at all. We conclude that alternative, behaviourally validated metrics for the visibility of flowers to pollinators, such as colour contrast and achromatic contrast, are better in understanding the role of flower colour in plant-pollinator signalling.

Evolution of Insect Color Vision: From Spectral Sensitivity to Visual Ecology.

Color vision is widespread among insects but varies among species, depending on the spectral sensitivities and interplay of the participating photoreceptors. The spectral sensitivity of a photoreceptor is principally determined by the absorption spectrum of the expressed visual pigment, but it can be modified by various optical and electrophysiological factors. For example, screening and filtering pigments, rhabdom waveguide properties, retinal structure, and neural processing all influence the perceived color signal. We review the diversity in compound eye structure, visual pigments, photoreceptor physiology, and visual ecology of insects. Based on an overview of the current information about the spectral sensitivities of insect photoreceptors, covering 221 species in 13 insect orders, we discuss the evolution of color vision and highlight present knowledge gaps and promising future research directions in the field.

Atmospheric H2S exposure does not affect stomatal aperture in maize.

MAIN CONCLUSION: Stomatal aperture in maize is not affected by exposure to a subtoxic concentration of atmospheric H2S. At least in maize, H2S, thus, is not a gaseous signal molecule that controls stomatal aperture. Sulfur is an indispensable element for the physiological functioning of plants with hydrogen sulfide (H2S) potentially acting as gasotransmitter in the regulation of stomatal aperture. It is often assumed that H2S is metabolized into cysteine to stimulate stomatal closure. To study the significance of H2S for the regulation of stomatal closure, maize was exposed to a subtoxic atmospheric H2S level in the presence or absence of a sulfate supply to the root. Similar to other plants, maize could use H2S as a sulfur source for growth. Whereas sulfate-deprived plants had a lower biomass than sulfate-sufficient plants, exposure to H2S alleviated this growth reduction. Shoot sulfate, glutathione, and cysteine levels were significantly higher in H2S-fumigated plants compared to non-fumigated plants. Nevertheless, this was not associated with changes in the leaf area, stomatal density, stomatal resistance, and transpiration rate of plants, meaning that H2S exposure did not affect the transpiration rate per stoma. Hence, it did not affect stomatal aperture, indicating that, at least in maize, H2S is not a gaseous signal molecule controlling this aperture.

Conical epidermal cells cause velvety colouration and enhanced patterning in Mandevilla flowers.

The majority of angiosperms have flowers with conical epidermal cells, which are assumed to have various functions, such as enhancing the visual signal to pollinators, but detailed optical studies on how conical epidermal cells determine the flower's visual appearance are scarce. Here we report that conical epidermal cells of Mandevilla sanderi flowers effectively reduce surface gloss and create a velvety appearance. Owing to the reduction in surface gloss, the flower further makes more efficient use of floral pigments and light scattering structures inside the flower. The interior backscattering yields a cosine angular dependence of reflected light, meaning that the flowers approximate near-perfect (Lambertian) diffusers, creating a visual signal that is visible across a wide angular space. Together with the large flowers and the tilted corolla tips, this generates a distinct visual pattern, which may enhance the visibility to pollinators.

Niche differentiation among clones in asexual grass thrips.

Many asexual animal populations comprise a mixture of genetically different lineages, but to what degree this genetic diversity leads to ecological differences remains often unknown. Here, we test whether genetically different clonal lineages of Aptinothrips grass thrips differ in performance on a range of plants used as hosts in natural populations. We find a clear clone-by-plant species interactive effect on reproductive output, meaning that clonal lineages perform differently on different plant species and thus are characterized by disparate ecological niches. This implies that local clonal diversities can be driven and maintained by frequency-dependent selection and that resource heterogeneity can generate diverse clone assemblies.

Vividly coloured poppy flowers due to dense pigmentation and strong scattering in thin petals.

The flowers of poppies (Papaveraceae) exhibit bright colours, despite their thin and floppy petals. We investigated the optical properties of flowers of Papaver rhoeas, P. dubium, Meconopsis cambrica and Argemone polyanthemos using a combined approach of anatomy, spectrophotometry and optical modelling. The petals of Papaver flowers are composed of only three cell layers, an upper and lower epidermal layer, which are densely filled with pigment, and an unpigmented mesophyll layer. Dense pigmentation together with strong scattering structures, composed of serpentine cell walls and air cavities, cause the striking poppy colours. We discuss how various aspects of the optical signal contribute to the flower's visibility to pollinators.

Colour preferences of Tetragonula carbonaria Sm. stingless bees for colour morphs of the Australian native orchid Caladenia carnea.

Innate colour preferences promote the capacity of pollinators to find flowers, although currently there is a paucity of data on how preferences apply to real flowers. The Australian sugarbag bee (Tetragonula carbonaria Sm.) has innate preferences for colours, including UV-absorbing white. Sugarbag bees are pollinators of the terrestrial orchid Caladenia carnea R.Br., which has both white and pink morphs. In laboratory conditions, we tested flower-naïve bees with the white and pink flower morphs revealing a significant preference for the white morph, consistent with experiments using artificial stimuli. In experiments to understand how bees may select food-deceptive orchids following habituation to a particular colour morph, we observed a significant increase in choices towards novel white flowers. We also observed that the presence of a UV-reflecting dorsal sepal signal significantly increased bee choices compared to flowers that had the UV signal blocked. Our findings demonstrate that innate preference testing of insect pollinators with artificial stimuli is replicated in a biologically significant scenario with flowers. The findings also underscore how food-deceptive orchids can receive sufficient pollinator visits to ensure pollination by having different morphs that draw on the innate preferences of bees and their ability to make decisions in a complex ecological setting.

The thermal ecology of flowers.

BACKGROUND: Obtaining an optimal flower temperature can be crucial for plant reproduction because temperature mediates flower growth and development, pollen and ovule viability, and influences pollinator visitation. The thermal ecology of flowers is an exciting, yet understudied field of plant biology. SCOPE: This review focuses on several attributes that modify exogenous heat absorption and retention in flowers. We discuss how flower shape, orientation, heliotropic movements, pubescence, coloration, opening-closing movements and endogenous heating contribute to the thermal balance of flowers. Whenever the data are available, we provide quantitative estimates of how these floral attributes contribute to heating of the flower, and ultimately plant fitness. OUTLOOK: Future research should establish form-function relationships between floral phenotypes and temperature, determine the fitness effects of the floral microclimate, and identify broad ecological correlates with heat capture mechanisms.

Functional significance of the optical properties of flowers for visual signalling.

Background: Flower coloration is a key enabler for pollinator attraction. Floral visual signals comprise several components that are generated by specific anatomical structures and pigmentation, and often have different functions in pollinator attraction. Anatomical studies have advanced our understanding of the optical properties of flowers, and evidence from behavioural experiments has elucidated the biological relevance of different components of floral visual signals, but these two lines of research are often considered independently. Scope: Here, we review current knowledge about different aspects of the floral visual signals, their anatomical and optical properties, and their functional significance in plant-pollinator visual signalling. We discuss common aspects, such as chromatic and achromatic contrast, hue, saturation and brightness, as well as less common types of visual signals, including gloss, fluorescence, polarization and iridescence in the context of salience of floral colour signals and their evolution, and highlight promising avenues for future research.

Pollinator or pedigree: which factors determine the evolution of pollen nutrients?

A prime example of plant-animal interactions is the interaction between plants and pollinators, which typically receive nectar and/or pollen as reward for their pollination service. While nectar provides mostly carbohydrates, pollen represents the main source of protein and lipids for many pollinators. However, the main function of pollen is to carry nutrients for pollen tube growth and thus fertilization. It is unclear whether pollinator attraction exerts a sufficiently strong selective pressure to alter the nutritional profile of pollen, e.g., through increasing its crude protein content or protein-to-lipid ratio, which both strongly affect bee foraging. Pollen nutritional quality may also be merely determined by phylogenetic relatedness, with pollen of closely related plants showing similar nutritional profiles due to shared biosynthetic pathways or floral morphologies. Here, we present a meta-analysis of studies on pollen nutrients to test whether differences in pollen nutrient contents and ratios correlated with plant insect pollinator dependence and/or phylogenetic relatedness. We hypothesized that if pollen nutritional content was affected by pollinator attraction, it should be different (e.g., higher) in highly pollinator-dependent plants, independent of phylogenetic relatedness. We found that crude protein and the protein-to-lipid ratio in pollen strongly correlated with phylogeny. Moreover, pollen protein content was higher in plants depending mostly or exclusively on insect pollination. Pollen nutritional quality thus correlated with both phylogenetic relatedness and pollinator dependency, indicating that, besides producing pollen with sufficient nutrients for reproduction, the nutrient profile of zoophilous plants may have been shaped by their pollinators' nutritional needs.

How to colour a flower: on the optical principles of flower coloration.

The coloration of flowers is due to the wavelength-selective absorption by pigments of light backscattered by structures inside the petals. We investigated the optical properties of flowers using (micro)spectrophotometry and anatomical methods. To assess the contribution of different structures to the overall visual signal of flowers, we used an optical model, where a petal is considered as a stack of differently pigmented and structured layers and we interpreted the visual signals of the model petals with insect vision models. We show that the reflectance depends, in addition to the pigmentation, on the petal's thickness and the inhomogeneity of its interior. We find large between-species differences in floral pigments, pigment concentration and localization, as well as floral interior structure. The fractions of reflected and transmitted light are remarkably similar between the studied species, suggesting common selective pressures of pollinator visual systems. Our optical model highlights that pigment localization crucially determines the efficiency of pigmentary filtering and thereby the chromatic contrast and saturation of the visual signal. The strongest visual signal occurs with deposition of pigments only on the side of viewing. Our systematic approach and optical modelling open new perspectives on the virtues of flower colour.

Coloration of the Chilean Bellflower, Nolana paradoxa, interpreted with a scattering and absorbing layer stack model.

MAIN CONCLUSION: An absorbing-layer-stack model allows quantitative analysis of the light flux in flowers and the resulting reflectance spectra. It provides insight in how plants can optimize their flower coloration for attracting pollinators. The coloration of flowers is due to the combined effect of pigments and light-scattering structures. To interpret flower coloration, we applied an optical model that considers a flower as a stack of layers, where each layer can be treated with the Kubelka-Munk theory for diffusely scattering and absorbing media. We applied our model to the flowers of the Chilean Bellflower, Nolana paradoxa, which have distinctly different-colored adaxial and abaxial sides. We found that the flowers have a pigmented, strongly scattering upper layer, in combination with an unpigmented, moderately reflecting lower layer. The model allowed quantitative interpretation of the reflectance and transmittance spectra measured with an integrating sphere. The absorbance spectrum of the pigment measured with a microspectrophotometer confirmed the spectrum derived by modeling. We discuss how different pigment localizations yield different reflectance spectra. The absorbing layer stack model aids in understanding the various constraints and options for plants to tune their coloration.

Iridescent flowers? Contribution of surface structures to optical signaling.

The color of natural objects depends on how they are structured and pigmented. In flowers, both the surface structure of the petals and the pigments they contain determine coloration. The aim of the present study was to assess the contribution of structural coloration, including iridescence, to overall floral coloration. We studied the reflection characteristics of flower petals of various plant species with an imaging scatterometer, which allows direct visualization of the angle dependence of the reflected light in the hemisphere above the petal. To separate the light reflected by the flower surface from the light backscattered by the components inside (e.g. the vacuoles), we also investigated surface casts. A survey among angiosperms revealed three different types of floral surface structure, each with distinct reflections. Petals with a smooth and very flat surface had mirror-like reflections and petal surfaces with cones yielded diffuse reflections. Petals with striations yielded diffraction patterns when single cells were illuminated. The iridescent signal, however, vanished when illumination similar to that found in natural conditions was applied. Pigmentary rather than structural coloration determines the optical appearance of flowers. Therefore, the hypothesized signaling by flowers with striated surfaces to attract potential pollinators presently seems untenable.

Visual ecology: How glossy colours shine and confuse.

Most colours in nature are matte, but across the tree of life glossiness has evolved numerous times, suggesting that glossiness can be beneficial. Recent research finds that glossiness may confuse observers and protect against predators.

Honesty, reliability, and information content of floral signals.

Plants advertise their presence by displaying attractive flowers, which pollinators use to locate a floral reward. Understanding how floral traits scale with reward status lies at the heart of pollination biology, because it connects the different interests of plants and pollinators. Studies on plant phenotype-reward associations often use different terms and concepts, which limits developing a broader synthesis. Here, we present a framework with definitions of the key aspects of plant phenotype-reward associations and provide measures to quantify them across different species and studies. We first distinguish between cues and signals, which are often used interchangeably, but have different meanings and are subject to different selective pressures. We then define honesty, reliability, and information content of floral cues/signals and provide ways to quantify them. Finally, we discuss the ecological and evolutionary factors that determine flower phenotype-reward associations, how context-dependent and temporally variable they are, and highlight promising research directions.

Mutualisms and (A)symmetry in Plant-Pollinator Interactions.

The majority of flowering plants relies on animal pollinators for sexual reproduction and many animal pollinators rely on floral resources. However, interests of plants and pollinators are often not the same, resulting in an asymmetric relationship that ranges from mutualistic to parasitic interactions. Our understanding of the processes that underlie this asymmetry remains fragmentary. In this Review, we bring together evidence from evolutionary biology, plant chemistry, biomechanics, sensory ecology and behaviour to illustrate that the degree of symmetry often depends on the perspective taken. We also highlight variation in (a)symmetry within and between plant and pollinator species as well as between geographic locations. Through taking different perspectives from the plant and pollinator sides we provide new ground for studies on the maintenance and evolution of animal pollination and on the (a)symmetry in plant-pollinator interactions.

Coloration of Flowers by Flavonoids and Consequences of pH Dependent Absorption.

Flavonoid pigments are key determinants of flower colors. As absorption spectra of flavonoids are known to be severely pH-dependent, cellular pH will play a crucial role in flower coloration. The flavonoids are concentrated in the vacuoles of the flowers' epidermal cells, and thus the pigments' absorption spectra are modulated by the vacuolar pH. Here we study the pH dependence of flavonoid absorption spectra in extracts from flowers of two poppy species Papaver dubium (red) and Meconopsis cambrica (orange), and a white and red Mandevilla sanderi variety. In the red poppy and Mandevilla flowers, absorption spectra of the cyanidin- and pelargonidin-based anthocyanins peak in the blue-green-wavelength range at low pH, but exhibit a distinct bathochromic shift at higher pH. This shift to longer wavelengths is not found for the blue-absorbing nudicaulin derivatives of M. cambrica, which have a similar absorption spectrum at low and high pH. The pH-dependent absorption changes of the white M. sanderi's flavonoid remained restricted to the UV. An analysis of the spectra with logistic functions suggests that the pH-dependent characteristics of the basic states of flavonols and anthocyanins are related. The implications of tuning of pH and pigment absorption spectra for studies on flower color evolution are discussed.

Mapping of Multiple Complementary Sex Determination Loci in a Parasitoid Wasp.

Sex determination has evolved in a variety of ways and can depend on environmental and genetic signals. A widespread form of genetic sex determination is haplodiploidy, where unfertilized, haploid eggs develop into males and fertilized diploid eggs into females. One of the molecular mechanisms underlying haplodiploidy in Hymenoptera, the large insect order comprising ants, bees, and wasps, is complementary sex determination (CSD). In species with CSD, heterozygosity at one or several loci induces female development. Here, we identify the genomic regions putatively underlying multilocus CSD in the parasitoid wasp Lysiphlebus fabarum using restriction-site associated DNA sequencing. By analyzing segregation patterns at polymorphic sites among 331 diploid males and females, we identify up to four CSD candidate regions, all on different chromosomes. None of the candidate regions feature evidence for homology with the csd gene from the honey bee, the only species in which CSD has been characterized, suggesting that CSD in L. fabarum is regulated via a novel molecular mechanism. Moreover, no homology is shared between the candidate loci, in contrast to the idea that multilocus CSD should emerge from duplications of an ancestral single-locus system. Taken together, our results suggest that the molecular mechanisms underlying CSD in Hymenoptera are not conserved between species, raising the question as to whether CSD may have evolved multiple times independently in the group.