George Karabourniotis

UV-B PROTECTIVE POTENTIAL AND FLAVONOID CONTENT OF LEAF HAIRS OF QUERCUS ILEX

Flavonoids of non-glandular leaf hairs from Quercus ilex were analysed. The main compounds were acylated kaempferol glycosides. Acylation shifted the absorption peak into the ultraviolet-B region of the spectrum in which intact trichome layers absorbed strongly. Ultraviolet-B radiation caused a considerable reduction of photosystem II photochemical efficiency only in dehaired leaves. It is suggested that leaf hairs, besides other roles, may function as an effective filter against the harmful ultraviolet-B radiation.

Beneficial effects of enhanced UV-B radiation under field conditions: improvement of needle water relations and survival capacity of Pinus pinea L. seedlings during the dry Mediterranean summer

The possible mechanism(s) by which supplemental UV-B radiation alleviates the adverse effects of summer drought in Mediterranean pines (Petropoulou et al. 1995) were investigated with seedlings of Pinus pinea. Plants received ambient or ambient plus supplemental UV-B radiation (biologically equivalent to a 15% ozone depletion over Patras, 38.3° N, 29.1° E) and natural precipitation or additional irrigation. Treatments started on 1 February, 1994 and lasted up to the end of the dry period (29 September). In well-watered plants, UV-B radiation had no influence on photosystem II photochemical efficiency and biomass accumulation. Water stressed plants suffered from needle loss and reduced photosystem II photochemical efficiency during the summer. These symptoms, however, were less pronounced in plants receiving supplemental UV-B radiation, resulting in higher total biomass at plant harvest. Laboratory tests showed that enhanced UV-B radiation did not improve the tolerance of photosystem II against drought, high light, high temperature and oxidative stress. Enhanced UV-B radiation, however, improved the water economy of water stressed plants, as judged by measurements of needle relative water content. In addition, it caused an almost two-fold increase of cuticle thickness. No such UV-B radiation effects were observed in well-watered pines. The results indicate that the combination of water stress and UV-B radiation may trigger specific responses, enabling the plants to avoid excessive water loss and, thereby, maintain a more efficient photosynthetic apparatus during the summer. The extent of this apparently positive UV-B radiation effect would depend on the amount of summer precipitation. Abbreviations: DW – dry weight, Fv/Fm – ratio of variable to maximum fluorescence, A 300 – absorbance at 300 nm, PAR – photosynthetically active radiation, PS II – photosystem II, RWC – relative water content, TCA – trichloroacetic acid, UV-BBE – biologically effective ultraviolet-B radiation

Trichome density and its UV-B protective potential are affected by shading and leaf position on the canopy

Abstract In Olea europaea trichome density and UV-B absorbing compounds of leaf hairs and the lamina proper of leaves located in south-facing, north-facing and the internal of the canopy were positively correlated to the UV-B midday instant irradiance measured in September at these three different positions of the canopy. The correlation between these three parameters and the receiving photosynthetically active radiation (PAR), however, was weaker. In Quercus ilex, trichome density and its UV-B absorbing capacity were considerably higher in the exposed, south-facing leaves, compared to the deeply shaded ones; the UV-B absorbing capacity of the de-haired lamina, however, was the same. In the broad-leaved, alpine rosette of Verbascum speciosum, one could distinguish two areas on the leaves, one exposed and one shaded by the superimposed lamina. Although trichome density and the UV-B absorbing compounds of the de-haired leaf were the same in the two areas, the UV-B absorbing capacity of hairs was considerably increased in the exposed region. In V. speciosum, exposure induced also qualitative changes in the UV-B absorbance profile, apparently due to the formation of new flavonoid compounds absorbing maximally at 345–350 nm. In all other cases, the differences were mainly quantitative. The results support the postulate of a function of leaf hairs as a UV-B radiation screen and suggest that trichome density and/or its UV-B absorbing capacity may depend on irradiance during leaf development.

The Relationship between Anatomy and Photosynthetic Performance of Heterobaric Leaves

Heterobaric leaves show heterogeneous pigmentation due to the occurrence of a network of transparent areas that are created from the bundle sheaths extensions (BSEs). Image analysis showed that the percentage of photosynthetically active leaf area (Ap) of the heterobaric leaves of 31 plant species was species dependent, ranging from 91% in Malva sylvestris to only 48% inGynerium sp. Although a significant portion of the leaf surface does not correspond to photosynthetic tissue, the photosynthetic capacity of these leaves, expressed per unit of projected area (Pmax), was not considerably affected by the size of their transparent leaf area (At). This means that the photosynthetic capacity expressed per Ap(P*max) should increase with At. Moreover, the expression of P*max could be allowing the interpretation of the photosynthetic performance in relation to some critical anatomical traits. The P*max, irrespective of plant species, correlated with the specific leaf transparent volume (λt), as well as with the transparent leaf area complexity factor (CFAt), parameters indicating the volume per unit leaf area and length/density of the transparent tissues, respectively. Moreover, both parameters increased exponentially with leaf thickness, suggesting an essential functional role of BSEs mainly in thick leaves. The results of the present study suggest that although the Ap of an heterobaric leaf is reduced, the photosynthetic performance of each areole is increased, possibly due to the light transferring capacity of BSEs. This mechanism may allow a significant increase in leaf thickness and a consequent increase of the photosynthetic capacity per unit (projected) area, offering adaptive advantages in xerothermic environments.

Leaf hairs of Olea europeae protect underlying tissues against ultraviolet-B radiation damage

Abstract The photochemical efficiency of photosystem II, as measured by chlorophyll fluorescence induction, was not affected in de-haired olive leaves kept in the dark or intact leaves irradiated with a moderate (3.75 W m −2 ) ultraviolet-B (UV-B) intensity. In de-haired, UV-B-irradiated leaves, however, the ratio of variable to maximum ( F v / F m ) chlorophyll fluorescence declined significantly and irreversibly. Reduction in F v / F m was associated with an increase in instantaneous ( F 0 ) and a decrease in maximum ( F m ) fluorescence, indicating perturbation by the UV-B exposure of more than one photosynthetic site. Extensive epidermal browning in dehaired, UV-B irradiated leaves was also observed, indicating possible damage to cell membranes. The results strengthen the hypothesis that leaf hairs protect the underlying tissues against UV-B radiation damage.

New Insights into the Properties of Pubescent Surfaces: Peach Fruit as a Model

The surface of peach (Prunus persica ‘Calrico’) is covered by a dense indumentum, which may serve various protective purposes. With the aim of relating structure to function, the chemical composition, morphology, and hydrophobicity of the peach skin was assessed as a model for a pubescent plant surface. Distinct physicochemical features were observed for trichomes versus isolated cuticles. Peach cuticles were composed of 53% cutan, 27% waxes, 23% cutin, and 1% hydroxycinnamic acid derivatives (mainly ferulic and p-coumaric acids). Trichomes were covered by a thin cuticular layer containing 15% waxes and 19% cutin and were filled by polysaccharide material (63%) containing hydroxycinnamic acid derivatives and flavonoids. The surface free energy, polarity, and work of adhesion of intact and shaved peach surfaces were calculated from contact angle measurements of water, glycerol, and diiodomethane. The removal of the trichomes from the surface increased polarity from 3.8% (intact surface) to 23.6% and decreased the total surface free energy chiefly due to a decrease on its nonpolar component. The extraction of waxes and the removal of trichomes led to higher fruit dehydration rates. However, trichomes were found to have a higher water sorption capacity as compared with isolated cuticles. The results show that the peach surface is composed of two different materials that establish a polarity gradient: the trichome network, which has a higher surface free energy and a higher dispersive component, and the cuticle underneath, which has a lower surface free energy and higher surface polarity. The significance of the data concerning water-plant surface interactions is discussed within a physiological context.

Polyphenol deposition in leaf hairs of Olea europaea (Oleaceae) and Quercus ilex (Fagaceae).

The subcellular localization (cytoplasm, vacuoles, cell walls) of polyphenol compounds during the development of the multicellular nonglandular leaf hairs of Olea europaea (scales) and Quercus ilex (stellates), was investigated. Hairs of all developmental stages were treated with specific inducers of polyphenol fluorescence, and the bright yellow-green fluorescence of individual hairs was monitored with epifluorescence microscopy. During the early ontogenetic stages, bright fluorescence was emitted from the cytoplasm of the cells composing the multicellular shield of the scales of O. europaea. Transmission electron micrographs of the same stages showed that these cells possessed poor vacuolation and thin cell walls. The nucleus of these cells may be protected against ultraviolet-B radiation damage. The progressive vacuolation that occurred during maturation was followed by a shifting of the bright green-yellow fluorescence from the perinuclear region and the cytoplasm to the cell walls. The same trends were observed during the development of the nonglandular stellate hairs of Quercus ilex, in which maturation was also accompanied by a considerable secondary thickening of the cell walls. Despite the differences in morphology, high concentrations of polyphenol compounds are initially located mainly in the cytoplasm of the developing nonglandular hairs, and their deposition on the cell walls takes place during the secondary cell wall thickening. These structural changes during the development of the leaf hairs make them a very effective barrier against abiotic (uv-B radiation) and probably biotic (pathogenic) stresses.

Wettability, Polarity, and Water Absorption of Holm Oak Leaves: Effect of Leaf Side and Age

The highly pubescent abaxial side of holm oak leaves is unwettable and water repellent, while the adaxial side is wettable and can take up water, which may be an adaptation to growing under Mediterranean conditions. Plant trichomes play important protective functions and may have a major influence on leaf surface wettability. With the aim of gaining insight into trichome structure, composition, and function in relation to water-plant surface interactions, we analyzed the adaxial and abaxial leaf surface of holm oak (Quercus ilex) as a model. By measuring the leaf water potential 24 h after the deposition of water drops onto abaxial and adaxial surfaces, evidence for water penetration through the upper leaf side was gained in young and mature leaves. The structure and chemical composition of the abaxial (always present) and adaxial (occurring only in young leaves) trichomes were analyzed by various microscopic and analytical procedures. The adaxial surfaces were wettable and had a high degree of water drop adhesion in contrast to the highly unwettable and water-repellent abaxial holm oak leaf sides. The surface free energy and solubility parameter decreased with leaf age, with higher values determined for the adaxial sides. All holm oak leaf trichomes were covered with a cuticle. The abaxial trichomes were composed of 8% soluble waxes, 49% cutin, and 43% polysaccharides. For the adaxial side, it is concluded that trichomes and the scars after trichome shedding contribute to water uptake, while the abaxial leaf side is highly hydrophobic due to its high degree of pubescence and different trichome structure, composition, and density. Results are interpreted in terms of water-plant surface interactions, plant surface physical chemistry, and plant ecophysiology.

“Carbon gain vs. water saving, growth vs. defence”: Two dilemmas with soluble phenolics as a joker

Despite that phenolics are considered as a major weapon against herbivores and pathogens, the primal reason for their evolution may have been the imperative necessity for their UV-absorbing and antioxidant properties in order for plants to compensate for the adverse terrestrial conditions. In dry climates the choice concerning the first dilemma (carbon gain vs. water saving) needs the appropriate structural and metabolic modulations, which protect against stresses such as high UV and visible radiation or drought, but reduce photosynthesis and increase oxidative pressure. Thus, when water saving is chosen, priority is given to protection (including phenolic synthesis), instead of carbon gain and hence growth. At the global level, the different choices by the individual species are expressed by an interspecific negative relationship between total phenolics and photosynthesis. On the other hand, the accumulation of phenolics in water saving plants offers additional defensive functions because these multifunctional compounds can also act as pro-oxidant, antifeeding or toxic factors. Therefore phenolics, as biochemical jokers, can give the answer to both dilemmas: water saving involves high concentrations of phenolics which also offer high level of defence.

Should structure-function relations be considered separately for homobaric vs. heterobaric leaves?

Tree and shrub species can be differentiated into two major groups based on their substantially different leaf anatomy: heterobaric and homobaric. In contrast to homobaric leaves, heterobaric leaves have bundle sheath extensions (BSEs) that create transparent regions on their lamina. Recent studies have shown that BSEs transfer visible light to internal mesophyll layers, thus affecting the photosynthetic performance of heterobaric leaves. Whether the two leaf types also differ in other functional and structural traits has not been addressed, nor have any structure-function relations. Here, we measured key anatomical and physiological parameters and tested their relationships in 30 species with different leaf types. Heterobaric leaves were thinner with lower leaf mass per area, had higher nitrogen concentration per mass, were (13)C-enriched, and achieved comparable photosynthetic capacity per area but had higher photosynthetic capacity per mass compared to homobaric leaves. Relations between leaf construction cost, nitrogen concentration, and photosynthesis followed the general pattern of the leaf economic spectrum, but differed between homobaric and heterobaric leaves. We suggest that the mechanisms controlling these relations differ between the two leaf types, presumably due to their distinct anatomy.