Cytoplasmic accumulation of flavonoids in flower petals and its relevance to yellow flower colouration

It is widely accepted that the mix of flavonoids in the cell vacuole is the source of flavonoid based petal colour, and that analysis of the petal extract reveals the nature and relative levels of vacuolar flavonoid pigments. However, it has recently been established with lisianthus flowers that some petal flavonoids can be excluded from the vacuolar mix through deposition in the cell wall or through complexation with proteins inside the vacuole, and that these flavonoids are not readily extractable. The present work demonstrates that flavonoids can also be compartmented within the cell cytoplasm. Using adaxial epidermal peels from the petals of lisianthus (Eustoma grandiflorum), Lathyrus chrysanthus and Dianthus caryophyllus, light and laser scanning confocal microscopy studies revealed a significant concentration of petal flavonoids in the cell cytoplasm of some tissues. With lisianthus, flavonoid analyses of isolated protoplasts and vacuoles were used to establish that ca 14% of petal flavonoids are located in the cytoplasm (cf. 30% in the cell wall and 56% in the vacuole). The cytoplasmic flavonoids are predominantly acylated glycosides (cf. non-acylated in the cell wall). Flavonoid aggregation on a cytoplasmic protein substrate provides a rational mechanism to account for how colourless flavonoid glycosides can produce yellow colouration in petals, and perhaps also in other plant parts. High vacuolar concentrations of such flavonoids are shown to be insufficient.     

The functional significance of colouration in cetaceans

Cetaceans show many of the classic mammalian colouration patterns, such as uniform colouration, countershading, and prominent patches of colour, all within one relatively small taxon. We collated all the functional hypotheses for cetacean colouration that have been put forward in the literature and systematically tested them using comparative phylogenetic analyses. We found that countershading is a mechanism by which smaller cetacean species may avoid being seen by their prey. We discovered that prominent markings are associated with group living, fast swimming, and ostentatious behaviour at the surface, suggesting that they function in intraspecific communication. White markings on several parts of the body seem to be involved in the capture of fish, squid, and krill. Therefore, several different selection pressures have shaped the great diversity of skin colouration seen in extant cetaceans, although background matching, disruptive colouration and interspecific communication do not appear to be involved.     

Anthocyanic vacuolar inclusions (AVIs) selectively bind acylated anthocyanins in Vitis vinifera L. (grapevine) suspension culture

Anthocyanic vacuolar inclusions (AVIs) appear as dark red-to-purple spheres of various sizes in vacuoles of grapevine (Vitis vinifera L.) cell suspension culture due to their interaction with anthocyanins. AVIs were purified and the bound anthocyanins extracted and analysed by HPLC from two lines of V. vinifera isolated from the same callus accumulating anthocyanin in the dark, yet varying in their anthocyanin profiles and accumulation. An intermediate-pigmented line (FU-1) with a 1.3:1 ratio of acylated:non-acylated anthocyanins, a colour value of 0.84 units and cyanidin and peonidin as the dominant species was compared with a high-pigmented line (FU-2) with a 1.2:1 ratio of acylated:non-acylated anthocyanins, a colour value of 3.72 units and malvidin predominating. The profile of AVI-bound anthocyanins showed an increase in acylated anthocyanins in both lines of approx. 28–29%, with no apparent preference for anthocyanin species. This resulted in a ratio of acylated:non-acylated anthocyanins of 6.2:1 for FU-1 and 4.9:1 for FU-2. The reasons for the selectivity of the AVIs for acylated (specifically p-coumaroylated) species compared with the whole cell profile are discussed.     

Characterisation of flower colouration in 30 Rhododendron species via anthocyanin and flavonol identification and quantitative traits

• Floral colour is a key reproductive character, often associated with environmental adaptation, and subject to human intervention. A large number of Rhododendron species differ widely in flower colour, providing a good model for flower colouration. The chromatic features and anthocyanin compositions of 30 species from seven subgenera were systematically analysed.
• The Royal Horticultural Society Colour Chart and CIE L*a*b* system were employed to describe and investigate flower colours. The UPLC‐PDA/ESI‐MSⁿ system was used to identify and quantify anthocyanins in petal extracts.
• The flower colours of 30 Rhododendron species were categorised into four groups – red, purplish pink, purple and white. Seven anthocyanins were identified and quantified in petals: delphinidin, cyanidin and malvidin 3‐O‐arabinoside‐5‐O‐glucosides, cyanidin 3,5‐di‐O‐glucoside, 3‐O‐galactoside and 3‐O‐arabinoside, and delphinidin 3‐O‐glucoside. The red‐flowered species mainly contained cyanidin monoglycosides and had much higher total anthocyanin content than purplish pink‐ and purple‐flowered species. Purplish pink‐ and purple‐flowered species had similar anthocyanin types and content. The chromatic differences were significant among groups, except the purplish pink and purple groups. Statistical analysis showed that Cy3Gal and Cy3Arb are characteristic for red‐flowered species, and Mv3Arb5G and Dp3Arb5G play important roles in purple colouration; their contents were major components that greatly affected the chromatic parameters. In total, 21 flavonol derivates were identified. However, total flavonol content and co‐pigmentation index showed no significant difference or correlation among/with colour groups, suggesting that flavonols might not play a major role in colouration.
• These results enhance our knowledge of the biochemical basis of flower colouration in Rhododendron species, and provide a foundation for genetic variation studies and aid in breeding cultivars with novel flower colours.

     

Genes encoding the vacuolar Na+/H+ exchanger and flower coloration

Vacuolar pH plays an important role in flower coloration: an increase in the vacuolar pH causes blueing of flower color. In the Japanese morning glory (Ipomoea nil or Pharbitis nil), a shift from reddish-purple buds to blue open flowers correlates with an increase in the vacuolar pH. We describe details of the characterization of a mutant that carries a recessive mutation in the Purple (Pr) gene encoding a vacuolar Na+/H+ exchanger termed InNHX1. The genome of I. nil carries one copy of the Pr (or InNHX1) gene and its pseudogene, and it showed functional complementation to the yeast nhx1 mutation. The mutant of I. nil, called purple (pr), showed a partial increase in the vacuolar pH during flower-opening and its reddish-purple buds change into purple open flowers. The vacuolar pH in the purple open flowers of the mutant was significantly lower than that in the blue open flowers. The InNHX1 gene is most abundantly expressed in the petals at around 12 h before flower-opening, accompanying the increase in the vacuolar pH for the blue flower coloration. No such massive expression was observed in the petunia flowers. Since the NHX1 genes that promote the transport of Na+ into the vacuoles have been regarded to be involved in salt tolerance by accumulating Na+ in the vacuoles, we can add a new biological role for blue flower coloration in the Japanese morning glory by the vacuolar alkalization.     

New insight into the structures and formation of anthocyanic vacuolar inclusions in flower petals

Background  

Although the biosynthetic pathways for anthocyanins and their regulation have been well studied, the mechanism of anthocyanin accumulation in the cell is still poorly understood. Different models have been proposed to explain the transport of anthocyanins from biosynthetic sites to the central vacuole, but cellular and subcellular information is still lacking for reconciliation of different lines of evidence in various anthocyanin sequestration studies. Here, we used light and electron microscopy to investigate the structures and the formation of anthocyanic vacuolar inclusions (AVIs) in lisianthus (Eustoma grandiflorum) petals.     

A new interpretation of flower guide colouration: Absorption of ultraviolet light enhances colour saturation

In melittophilous plants the colour pattern of the flowers, as perceived by bumblebees, is a gradient of centripetally increasing spectral purity. This pattern serves as a signal for innate flower recognition in naive bumblebees permitting orientation to flowers and landing on flowers. Structures which make up the total signal pattern can include the background (e.g., green leaves), corollas, and stamens or floral guides. How various colour parameters, such as dominant wavelength, intensity, and spectral purity influence the colour signal pattern of flowers is analyzed. The process of strong absorption of ultraviolet light is shown to be a mechanism for the enhancement of spectral purity in flower guides. The importance of other mechanisms is also demonstrated. The presence of a gradient of centripetally increasing spectral purity in floral colour patterns as perceived by a bumblebee's eyes is demonstrated by a comparison of the spectral reflectance in different parts of the flower and a representation of colour loci in the colour triangle.     

Sexual colouration and sperm traits in guppies

The relationships among the area, hue, saturation and brightness of orange colouration and sperm traits in the guppy Poecilia reticulata were investigated. Males with greater areas of orange colouration had significantly larger sperm loads, more motile sperm and longer sperm relative to males with relatively little orange colouration. Males with greater areas of orange colouration did not possess more viable sperm than males with relatively little orange colouration. There was no evidence that any of the sperm traits were related to the hue, saturation or brightness of the orange colouration. These results are discussed in the context of the roles that direct and indirect selection might play in maintaining female preference for male guppies with large areas of orange colouration.     

Spondylolysis: its nature and anthropological significance

Spondylolysis of the lower back is a kind of vertebral fracture that is unique to humans being related to anatomical features such as the lumbar curve which facilitate upright posture and bipedal locomotion in this species. It is virtually never seen in children before the age of walking and has not been observed in adults who were never able to walk. Although apparently produced on occasion by acute trauma, spondylolysis usually develops as a fatigue fracture in response to sustained or repeated stress. The occurrence of spondylolysis varies greatly from population to population with the highest frequencies, approaching 50%, seen in northern Alaskan and Greenlandic Eskimos. Males are affected more frequently than females, and the condition is common in athletes, particularly gymnasts, divers, polevaulters, and American football players. It is also seen among heavy laborers, especially if the job requires lifting. Although generally approached as a pathological condition requiring corrective treatment, spondylolysis, as well as the anterior slippage (olisthesis) that frequently accompanies it, usually produces relatively mild symptoms if any at all. In fact, it many even provide advantages such as greater flexibility in the lower back.     

Do nectar feeders in Andean nature reserves affect flower visitation by hummingbirds?

The interactions among plants and pollinators are crucial for plant reproduction because they structure gene flow among individual plants and among populations of plants. Throughout the Neotropics there has been a strong increase in recent years in the use of artificial nectar feeders in private nature reserves to encourage ecotourism through prolonged observations of hummingbirds. Currently, there is considerable uncertainty whether artificial feeders have a detrimental effect on plant reproduction through competition or a beneficial effect through facilitation. This uncertainty is disconcerting given that nature reserves harbour many rare and endangered plants whose successful reproduction is a conservation goal. To assess whether nectar feeders affected hummingbird visitation to flowering plants, we determined visitation rates in ten flowering species in five Andean nature reserves in Ecuador. We found that visitation rates tended to be higher within 5 m around the feeders than they were at 100 m, 500 m, or 1.5 km distance to the feeders, therefore indicating that nectar feeders tended to facilitate flower visitation at close distance. Because visitation rates at 100 m and 500 m distance from the feeders did not differ from those at 1.5 km, we suggest that feeders do not draw hummingbirds away from flower resources, but if they have an effect on flower visitation at all, they tend to facilitate flower visitation rather than reduce plant reproduction.     

The significance of flower colour change in eight co-occurring shrub species

Flower colour changes from white or yellow to various shades of red at or near the sites of harvestable pollen in Calytrix glutinosa, Grevillea pilulifera, Isopogon dubius and Petrophile biloba , and over most of the flower in Hypocalymma angustifolium, Verticordia chrysantha and V. huegelii and over the pseudanthium in Darwinia citriodora. All bee, wasp, beetle, fly, butterfly and moth visitors select flowers in the white/yellow phase rather than the red or intermediate phase.
Nectar is produced by five species, harvestable pollen by four species and detectable perfume by three species, all of which features are usually absent from the red phase. The timing of the colour change in all species also corresponds to loss of stigma receptivity, completion of pollination and onset of ovule seed) swelling. Six species also undergo minor morphometric changes which discourage visitation. In all species, colour change is non-inducible by pollinators, taking 2–30 days to complete. In three protandrous species, all available pollen may be removed in the first visit, requiring transport of non-self pollen to rewardless flowers during the 10 h period of the yellow phase.
These species are highly floriferous and occur in dense patches. Since only a small proportion of flowers may be receptive at any one time, it is concluded that retention of flower parts essentially serves to enhance long-distance attraction, while colour change maximizes pollination and foraging efficiency.     

Size-dependent colouration balances conspicuous aposematism and camouflage

Colour is an important component of many different defensive strategies, but signal efficacy and detectability will also depend on the size of the coloured structures, and how pattern size interacts with the background. Consequently, size-dependent changes in colouration are common among many different species as juveniles and adults frequently use colour for different purposes in different environmental contexts. A widespread strategy in many species is switching from crypsis to conspicuous aposematic signalling as increasing body size can reduce the efficacy of camouflage, while other antipredator defences may strengthen. Curiously, despite being chemically defended, the gold-striped frog (Lithodytes lineatus, Leptodactylidae) appears to do the opposite, with bright yellow stripes found in smaller individuals, whereas larger frogs exhibit dull brown stripes. Here, we investigated whether size-dependent differences in colour support distinct defensive strategies. We first used visual modelling of potential predators to assess how colour contrast varied among frogs of different sizes. We found that contrast peaked in mid-sized individuals while the largest individuals had the least contrasting patterns. We then used two detection experiments with human participants to evaluate how colour and body size affected overall detectability. These experiments revealed that larger body sizes were easier to detect, but that the colours of smaller frogs were more detectable than those of larger frogs. Taken together our data support the hypothesis that the primary defensive strategy changes from conspicuous aposematism to camouflage with increasing size, implying size-dependent differences in the efficacy of defensive colouration. We discuss our data in relation to theories of size-dependent aposematism and evaluate the evidence for and against a possible size-dependent mimicry complex with sympatric poison frogs (Dendrobatidae).     

Cytomixis, its nature, significance and the cytological consequences

Cytomixis is the widespread natural process of intercellular interaction which is characteristic for vegetative and generative tissues in both normal and pathological conditions. The origin significance and genetic control cytomixis still remain not completely clear. The popularity of view of the pathological nature of cytomixis based on its peculiar plants with genetic instability and impaired homeostasis. In the genetic control of cytomixis seem to be involved meiotic genes which are responsible for segregation and organization of chromosomes. Their activity is modified by environmental factors through signal transduction. It is assumed via cytomixis, from one side, the informational contact can be reached and meiosis and gametogenesis are synchronized, with another, increase of the genetic variety and level of the heterozygosis of microsporocytes. The activity of cytomixis varies over wide limits. The greatest influence on its activity have mutagenesis hybridization and polyploidy. In this context cytomixis can fulfill the function of cell selection which is activated by exceeding of the threshold level of the microsporocyte damages (or genetical disbalance).     

Background colouration influences body colour segregation in mollies

When given a choice of swimming against a black background near an empty black end compartment or swimming against a white background near an empty white compartment, black morph mollies Poecilia latipinna spent significantly more time against the black background. The presence of a shoal (either white or black mollies) in the black compartment increased the preference of the test fish for the black background. However, black molly test fish spent significantly less time against a black background if a black shoal was present in the white compartment than in assays in which a white shoal was in the white compartment. In assays where test fish were given the choice between two black shoals, test fish spent significantly more time near the black shoal on the black background. Similarly, when given the choice between two white shoals, test fish spent significantly more time against the black background. The results suggest that the presence of cryptic backgrounds alter shoalmate choices in fish that assort according to body colouration.