Disentangling leaf structural and material properties in relationship to their anatomical and chemical compositional traits in oaks (Quercus L.).

BACKGROUND AND AIMS: The existence of sclerophyllous plants has been considered an adaptive strategy against different environmental stresses. Given that it literally means 'hard-leaved', it is essential to quantify the leaf mechanical properties to understand sclerophylly. However, the relative importance of each leaf trait for mechanical properties is not yet well established. METHODS: Genus Quercus is an excellent system to shed light on this because it minimizes phylogenetic variation while having a wide variation in sclerophylly. We measured leaf anatomical traits and cell wall composition, analysing their relationship with leaf mass per area and leaf mechanical properties in a set of 25 oak species. KEY RESULTS: The upper epidermis outer wall makes a strong and direct contribution to the leaf mechanical strength. Moreover, cellulose plays a crucial role in increasing leaf strength and toughness. The principal component analysis plot based on leaf trait values clearly separates Quercus species into two groups corresponding to evergreen and deciduous species. CONCLUSIONS: Sclerophyllous Quercus species are tougher and stronger owing to their thicker epidermis outer wall and/or higher cellulose concentration. Furthermore, section Ilex species share common traits, although they occupy different climates. In addition, evergreen species living in mediterranean-type climates share common leaf traits irrespective of their different phylogenetic origin.

Cell-level anatomical characteristics explain high mesophyll conductance and photosynthetic capacity in sclerophyllous Mediterranean oaks.

Leaf mass per area (LMA) has been suggested to negatively affect the mesophyll conductance to CO2 (gm ), which is the most limiting factor for area-based photosynthesis (AN ) in many Mediterranean sclerophyll species. However, despite their high LMA, these species have similar AN to plants from other biomes. Variations in other leaf anatomical traits, such as mesophyll and chloroplast surface area exposed to intercellular air space (Sm /S and Sc /S), may offset the restrictions imposed by high LMA in gm and AN in these species. Seven sclerophyllous Mediterranean oaks from Europe/North Africa and North America with contrasting LMA were compared in terms of morphological, anatomical and photosynthetic traits. Mediterranean oaks showed specific differences in AN that go beyond the common morphological leaf traits reported for these species (reduced leaf area and thick leaves). These variations resulted mainly from the differences in gm , the most limiting factor for carbon assimilation in these species. Species with higher AN showed increased Sc /S, which implies increased gm without changes in stomatal conductance. The occurrence of this anatomical adaptation at the cell level allowed evergreen oaks to reach AN values comparable to congeneric deciduous species despite their higher LMA.

Evolutionary divergence of leaf width and its correlates.

UNLABELLED: • PREMISE OF THE STUDY: The question why leaf dimensions vary so much between species has long puzzled ecologists. Presumably, variation arises from selective forces acting on leaf function but which selective forces and which leaf functions? This investigation assesses the consistency of divergence in plant traits and habitat variables in association with leaf width divergence in the flora of NSW, Australia.• METHODS: More than 80 traits and habitat variables were measured for 25 independent evolutionary divergence events (PICs). Each PIC was represented by two related plant species that had diverged substantially in leaf width. Outgroup species provided indications of the direction of divergence. Most PICs were within genus, so divergences represent relatively recent evolutionary events.• KEY RESULTS: No plant traits or habitat variables were 100% consistently associated with a divergence in leaf width, and surprisingly few diverged in a consistent direction significantly more than what might be expected by chance. This surprising lack of consistent divergence with leaf width contrasted with the result that many of these traits and habitat variables were correlated with leaf width across all species in our data set and in line with correlations reported from other studies. Subcategorizing PICs according to the probable direction of leaf width divergence did not improve consistency.• CONCLUSIONS: These results indicate that evolutionarily recent leaf width divergence events are not tightly tied to divergences in other leaf traits or in environmental situations, despite the broad correlations that have been observed across many species. Rather, cross species correlations are underpinned by earlier divergence events in the phylogeny.

Early evidence of xeromorphy in angiosperms: stomatal encryption in a new eocene species of Banksia (Proteaceae) from Western Australia.

UNLABELLED: • PREMISE OF THE STUDY: Globally, the origins of xeromorphic traits in modern angiosperm lineages are obscure but are thought to be linked to the early Neogene onset of seasonally arid climates. Stomatal encryption is a xeromorphic trait that is prominent in Banksia, an archetypal genus centered in one of the world's most diverse ecosystems, the ancient infertile landscape of Mediterranean-climate southwestern Australia.• METHODS: We describe Banksia paleocrypta, a sclerophyllous species with encrypted stomata from silcretes of the Walebing and Kojonup regions of southwestern Australia dated as Late Eocene.• KEY RESULTS: Banksia paleocrypta shows evidence of foliar xeromorphy ∼20 Ma before the widely accepted timing for the onset of aridity in Australia. Species of Banksia subgenus Banksia with very similar leaves are extant in southwestern Australia. The conditions required for silcrete formation infer fluctuating water tables and climatic seasonality in southwestern Australia in the Eocene, and seasonality is supported by the paucity of angiosperm closed-forest elements among the fossil taxa preserved with B. paleocrypta. However, climates in the region during the Eocene are unlikely to have experienced seasons as hot and dry as present-day summers.• CONCLUSIONS: The presence of B. paleocrypta within the center of diversity of subgenus Banksia in edaphically ancient southwestern Australia is consistent with the continuous presence of this lineage in the region for ≥40 Ma, a testament to the success of increasingly xeromorphic traits in Banksia over an interval in which numerous other lineages became extinct.

Diversity of Gall-Inducing Insects Associated With a Widely Distributed Tropical Tree Species: Testing the Environmental Stress Hypothesis.

Abiotic factors can affect plant performance and cause stress, which in turn affects plant-herbivore interactions. The Environmental Stress Hypothesis (ESH) predicts that gall-inducing insect diversity will be greater on host plants that grow in stressful habitats. We tested this hypothesis, considering both historical and ecological scales, using the plant Copaifera langsdorffii Desf. (Fabaceae) as a model because it has a wide geographic distribution and is a super-host of gall-inducing insects. According to the ESH, we predicted that 1) on a historical scale, the diversity of gall-inducing insects will be higher in habitats with greater environmental stress and 2) on an ecological scale, gall-inducing insect diversity will be greater on plants that possess greater levels of foliar sclerophylly. We sampled gall-inducing insects on plants of C. langsdorffii in five sites with different levels of water and soil nutrient availability and separated from each other by a distance of up to 470 km. The composition, richness, and abundance of gall-inducing insects varied among study sites. Plants located in more stressful habitats had higher levels of foliar sclerophylly; but richness and abundance of gall-inducing insects were not affected by host plant sclerophylly. Habitat stress was a good predictor of gall-inducing insect diversity on a regional scale, thus corroborating the first prediction of the ESH. No relationship was found between plant sclerophylly and gall-inducing insect diversity within habitats. Therefore, on a local scale, we did not find support for our second prediction related to the ESH.

Acclimation of Bryophytes to Sun Conditions, in Comparison to Shade Conditions, Is Influenced by Both Photosynthetic and Ultraviolet Radiations.

We studied the acclimation modalities of bryophytes to sun and shade under ambient or close-to-ambient conditions, measuring variables usually influenced by photosynthetically active (PAR) and ultraviolet (UV) radiations. Our aim was to elucidate to what extent the responses to changing radiations were influenced by PAR and UV wavelengths. For this aim, we used three taxonomically and structurally different species: the thalloid liverwort Marchantia polymorpha subsp. polymorpha, the leafy liverwort Jungermannia exsertifolia subsp. cordifolia, and the moss Fontinalis antipyretica. In the field, liverworts were more radiation-responsive than the moss, and the thalloid liverwort was more responsive than the leafy liverwort. Sun plants of M. polymorpha showed, in comparison to shade plants, higher sclerophylly, lower Chl a + b contents, higher Chl a/b ratios, higher (antheraxanthin + zeaxanthin)/(violaxanthin + antheraxanthin + zeaxanthin) ratios (xanthophyll index), lower F v/F m values, higher contents of methanol-soluble vacuolar UV-absorbing compounds (soluble UVACs), higher values of the ratio between the contents of methanol-insoluble cell wall-bound UVACs (insoluble UVACs) and soluble UVACs, higher contents of soluble luteolin and apigenin derivatives and riccionidin A, and higher contents of insoluble p-coumaric and ferulic acids. Overall, these responses reduced light absorption, alleviated overexcitation, increased photoprotection through non-photochemical energy dissipation, increased UV protection through UV screening and antioxidant capacity, and denoted photoinhibition. J. exsertifolia showed moderate differences between sun and shade plants, while responses of F. antipyretica were rather diffuse. The increase in the xanthophyll index was the most consistent response to sun conditions, occurring in the three species studied. The responses of soluble UVACs were generally clearer than those of insoluble UVACs, probably because insoluble UVACs are relatively immobilized in the cell wall. These modalities of radiation acclimation were reliably summarized by principal components analysis. Using the most radiation-responsive species in the field (M. polymorpha), we found, under close-to-ambient greenhouse conditions, that sclerophylly and Chl a + b content were only influenced by PAR, F v/F m, and luteolin and apigenin derivatives were only determined by UV, and xanthophyll index was influenced by both radiation types. Thus, responses of bryophytes to radiation can be better interpreted considering the influence of both PAR and UV radiation.

Leaf traits in Chilean matorral: sclerophylly within, among, and beyond matorral, and its environmental determinants.

Studies of leaf traits often focus on tradeoffs between growth and resource conservation, but little is known about variation in the mechanical traits that influence resource conservation. This study investigates how leaf mechanical traits vary across matorral vegetation in central Chile, how they correlate with environmental factors, and how these trends compare at a broader geographic scale. Leaf toughness, strength, stiffness, and associated traits were measured in five matorral types in central Chile, and relationships with soil N and P and climate variables were assessed. Trends with soil and climate were then analyzed across shrubland and woodland in Chile, Western Australia, and New Caledonia. Chilean species varied in leaf mechanics and associated traits, both within and among matorral types, with more species in sclerophyll matorral having strong, tough, and stiff leaves than in arid and littoral matorral. Overall, leaves with high leaf dry mass per area were stiffer, tougher, stronger, thicker, denser, with more fiber, lignin, phenolics and fiber per unit protein and less protein: tannin activity and N and P per mass, forming a broad sclerophylly syndrome. Mechanical traits of matorral species were not correlated with soil N or P, or predictably with climate variables, except flexural stiffness (EI W) which correlated positively with annual reference evapotranspiration (ET 0). However, soil P made strong independent contributions to variation in leaf mechanics across shrublands and woodlands of Chile, Western Australia, and New Caledonia, either separately (strength) or together with ET 0 (toughness) explaining 46-90% of variation. Hence ET 0 was predictive of EI W in Chilean matorral, whereas soil P was highly predictive of variation in leaf strength, and combined with ET 0 was highly predictive of toughness, at a broader geographic scale. The biological basis of these relationships, however, may be complex.

Different strategies of drought resistance in three Mediterranean sclerophyllous trees growing in the same environmental conditions.

Measurements have been made of leaf conductance to water vapour, relative water content and water potential in Olea oleaster Hoffmgg et Link, Ceratonia siliqua L. and Laurus nobilis L., three evergreen sclerophyllous trees growing in Sicily at sea level. Measurements were made hourly in May and September 1986. Although all the three species are regarded as components of a homogeneous group (sclerophylls) and they all showed a high degree of sclerophylly, the strategies they adopted to withstand drought were completely different. Olea oleaster behaved as a 'drought-tolerating' species. Drought was 'avoided' by Ceratonia siliqua by a 'water-spending' strategy and by Laurus nobilis by a 'water-saving' strategy combined with the capability of recovering even minimal water losses by dropping leaf water potential drastically. Such differences in drought resistance are discussed in terms of differences in the bulk elastic modulus of leaf cells and wood anatomy.

Characterizing sclerophylly: the mechanical properties of a diverse range of leaf types.

• Although sclerophylly is defined by textural properties, its adaptive significance has been debated without a strong base of mechanical data. We measured a wide range of mechanical properties across a diverse range of species and leaf forms, including highly scleromorphic leaves, and compared these with sclerophylly indices to determine the mechanical properties of sclerophylls. • Fracture and flexure tests were used to determine leaf strength, toughness (work to fracture) and flexural stiffness ('structural' properties), and specific strength, specific toughness and Young's modulus of elasticity ('material' properties, i.e. normalized per unit leaf thickness). • Leaves varied considerably in all properties tested, and in the way they combined various 'structural' and 'material' properties. However, on average, highly scleromorphic leaves were stronger, tougher and stiffer than soft leaves. 'Structural' properties correlated more strongly with sclerophylly than 'material' properties, and the ratio of stiffness to strength and toughness increased in sclerophyllous species. • Of the structural properties, strength, toughness and flexural stiffness each made substantial independent contributions to the variation in sclerophylly indices, but the best individual explanators were flexural stiffness and strength, with the best predictive model being a combination of these two properties. This model should now be tested on leaves from contrasting environments.

Leaves at low versus high rainfall: coordination of structure, lifespan and physiology.

• Across species, leaf lifespan (LL) tends to be correlated with leaf mass per area (LMA). Previously we found that Australian perennial species from low-rainfall sites had c . 40% shorter LL at a given LMA than high-rainfall species. • Here we relate indices of leaf strength (work to shear, W shear , and tissue toughness) to LL and LMA across the same suite of species. W shear is the work required to cut a leaf with a blade; W shear divided by leaf thickness gives tissue toughness. • Low- and high-rainfall species did not differ in their LL at a given W shear , but dry-site species had lower W shear at a given LMA, leading to the observed LL - LMA shift with rainfall. These patterns were driven by 50% lower tissue toughness in dry-site species. • The lower toughness was linked with high leaf N concentration, which is known to enhance water conservation during photosynthesis in low-rainfall species. Our results suggest that a significant cost of this strategy is reduced LL for a given investment in leaf tissue (LMA).

Characterising sclerophylly: some mechanical properties of leaves from heath and forest.

Although sclerophylly is widespread through the world and is often the dominant leaf-form in mediterranean climates, the mechanical properties of sclerophyllous leaves are poorly understood. The term "sclerophyllous" means hard-leaved, but biologists also use terms such as tough, stiff and leathery to describe sclerophyllous leaves. The latter term has no precise definition that allows quantification. However, each of the former terms is well-defined in materials engineering, although they may be difficult or sometimes inappropriate to measure in leaves because of their size, shape or composite and anisotropic nature. Two of the most appropriate and practically applicable mechanical properties of sclerophyllous leaves are "strength" and "toughness", which in this study were applied using punching, tearing and shearing tests to 19 species of tree and shrub at Wilson's Promontory, Australia. The results of these tests were compared with leaf specific mass (LSM) and a sclerophylly index derived from botanists' ranks. Principal components analysis was used to reduce the set of mechanical properties to major axes of variation. Component 1 correlated strongly with the botanists' ranks. Overall, leaves ranked as sclerophyllous by botanists were both tough and strong in terms of punching and tearing tests. In addition, tough and strong leaves typically had high toughness and strength per unit leaf thickness. There was also a significant correlation between component 1 and LSM. Although more detailed surveys are required, we argue that sclerophylly should be defined in terms of properties that have precise meanings and are measurable, such as toughness and strength, and that relate directly to mechanical properties as implicit in the term.

Leaf specific mass confounds leaf density and thickness.

We explored the relationship between leaf specific mass (LSM) and its two components, leaf density and thickness. These were assessed on the leaves of (a) the moderately sclerophyllous tree Arbutus menziesii distributed along a natural nutrient/moisture gradient in California, (b) eight sclerophyllous shrub species on four substrates in south-western Australia, and (c) seedlings of two morphologically contrasting Hakea species grown under varying soil nutrient, moisture and light regimes in a glasshouse experiment. Leaf area, mass, LSM, density and thickness varied greatly between leaves on the same plant, different species, and with different nutrient, moisture and light regimes. In some cases, variations in LSM were due to changes in leaf density in particular or thickness or both, while in others, density and thickness varied without a net effect on LSM. At lower nutrient or moisture availabilities or at higher light irradiances, leaves tended to be smaller, with higher LSM, density and thickness. Under increased stress, the thickness (diameter) of needle leaves decreased despite an increase in LSM. We concluded that, while LSM is a useful measure of sclerophylly, its separation into leaf density and thickness may be more appropriate as they often vary independently and appear to be more responsive to environmental gradients than LSM.