Local adaptation optimizes photoprotection strategies in a Neotropical legume tree under drought stress.

Photoprotection is a plant functional mechanism to prevent photooxidative damage by excess light. This is most important when carbon assimilation is limited by drought, and as such, it entails a trade-off between carbon assimilation vs stress avoidance. The ecological adaptation of plants to local water availability can lead to different photoprotective strategies. To test this, we used different provenances of Caesalpinia spinosa (Mol.) Kuntze (commonly known as 'tara') along a precipitation gradient. Tara is a Neotropical legume tree with high ecological and commercial value, found in dry tropical forests, which are increasingly threatened by climate change. Morphological and physiological responses of tara provenances were analysed under three different treatments of drought and leaflet immobilization, i.e., light stress, in a common garden greenhouse experiment. Tara quickly responded to drought by reducing stomatal conductance, evapotranspiration, photochemical efficiency, carbon assimilation and growth, while increasing structural and chemical photoprotection (leaflet angle and pigments for thermal dissipation). Leaflet closure was an efficient photoprotection strategy with overall physiological benefits for seedlings as it diminished the evaporative demand and avoided photodamage, but also entailed costs by reducing net carbon assimilation opportunities. These responses depended on seed origin, with seedlings from the most xeric locations showing the highest dehydration tolerance, suggesting local adaptation and highlighting the value of different strategies under distinct environments. This plasticity in its response to environmental stress allows tara to thrive in locations with contrasting water availability. Our findings increase the understanding of the factors controlling the functional ecology of tara in response to drought, which can be leveraged to improve forecasts of changes in its distribution range, and for planning restoration projects with this keystone tree species.

Unusual positional effects on flower sex in an andromonoecious tree: Resource competition, architectural constraints, or inhibition by the apical flower?

PREMISE OF THE STUDY: Two, nonmutually exclusive, mechanisms-competition for resources and architectural constraints-have been proposed to explain the proximal to distal decline in flower size, mass, and/or femaleness in indeterminate, elongate inflorescences. Whether these mechanisms also explain unusual positional effects such as distal to proximal declines of floral performance in determinate inflorescences, is understudied. METHODS: We tested the relative influence of these mechanisms in the andromonoecious wild olive tree, where hermaphroditic flowers occur mainly on apical and the most proximal positions in determinate inflorescences. We experimentally increased the availability of resources for the inflorescences by removing half of the inflorescences per twig or reduced resource availability by removing leaves. We also removed the apical flower to test its inhibitory effect on subapical flowers. KEY RESULTS: The apical flower had the highest probability of being hermaphroditic. Further down, however, the probability of finding a hermaphroditic flower decreased from the base to the tip of the inflorescences. An experimental increase of resources increased the probability of finding hermaphroditic flowers at each position, and vice versa. Removal of the apical flower increased the probability of producing hermaphroditic flowers in proximal positions but not in subapical positions. CONCLUSIONS: These results indicate an interaction between resource competition and architectural constraints in influencing the arrangement of the hermaphroditic and male flowers within the inflorescences of the wild olive tree. Subapical flowers did not seem to be hormonally suppressed by apical flowers. The study of these unusual positional effects is needed for a general understanding about the functional implications of inflorescence architecture.

The expression of light-related leaf functional traits depends on the location of individual leaves within the crown of isolated Olea europaea trees.

BACKGROUND: The spatial arrangement and expression of foliar syndromes within tree crowns can reflect the coupling between crown form and function in a given environment. Isolated trees subjected to high irradiance and concomitant stress may adjust leaf phenotypes to cope with environmental gradients that are heterogeneous in space and time within the tree crown. The distinct expression of leaf phenotypes among crown positions could lead to complementary patterns in light interception at the crown scale. METHODS: We quantified eight light-related leaf traits across 12 crown positions of ten isolated Olea europaea trees in the field. Specifically, we investigated whether the phenotypic expression of foliar traits differed among crown sectors and layers and five periods of the day from sunrise to sunset. We investigated the consequences in terms of the exposed area of the leaves at the tree scale during a single day. KEY RESULTS: All traits differed among crown positions except the length-to-width ratio of the leaves. We found a strong complementarity in the patterns of the potential exposed area of the leaves among day periods as a result of a non-random distribution of leaf angles across the crown. Leaf exposure at the outer layer was below 60 % of the displayed surface, reaching maximum interception during morning periods. Daily interception increased towards the inner layer, achieving consecutive maximization from east to west positions within the crown, matching the sun's trajectory. CONCLUSIONS: The expression of leaf traits within isolated trees of O. europaea varies continuously through the crown in a gradient of leaf morphotypes and leaf angles depending on the exposure and location of individual leaves. The distribution of light-related traits within the crown and the complementarity in the potential exposure patterns of the leaves during the day challenges the assumption of low trait variability within individuals.

The effects of phenotypic plasticity and local adaptation on forecasts of species range shifts under climate change.

Species are the unit of analysis in many global change and conservation biology studies; however, species are not uniform entities but are composed of different, sometimes locally adapted, populations differing in plasticity. We examined how intraspecific variation in thermal niches and phenotypic plasticity will affect species distributions in a warming climate. We first developed a conceptual model linking plasticity and niche breadth, providing five alternative intraspecific scenarios that are consistent with existing literature. Secondly, we used ecological niche-modeling techniques to quantify the impact of each intraspecific scenario on the distribution of a virtual species across a geographically realistic setting. Finally, we performed an analogous modeling exercise using real data on the climatic niches of different tree provenances. We show that when population differentiation is accounted for and dispersal is restricted, forecasts of species range shifts under climate change are even more pessimistic than those using the conventional assumption of homogeneously high plasticity across a species' range. Suitable population-level data are not available for most species so identifying general patterns of population differentiation could fill this gap. However, the literature review revealed contrasting patterns among species, urging greater levels of integration among empirical, modeling and theoretical research on intraspecific phenotypic variation.

Leaf-trait responses to irrigation of the endemic fog-oasis tree Myrcianthes ferreyrae: can a fog specialist benefit from regular watering?

Myrcianthes ferreyrae is an endemic, endangered species, with a small number of individuals located only in hyperarid, fog-oases known as lomas along the Peruvian desert in southern Peru, where fog is the main source of water. Following centuries of severe deforestation, reforestation with this native species was conducted in the Atiquipa lomas, Arequipa-Perú. On five slopes, five 2-year-old seedlings were irrigated monthly with water trapped by raschel-mesh fog collectors, supplementing natural rainfall with 0, 20, 40, 60 and 80 mm month(-1) from February to August 2008. We measured plant growth, increment in basal diameter, height and five leaf traits: leaf mass area (LMA), leaf carbon isotope composition (δ(13)C), nitrogen per leaf area, total leaf carbon and stomatal density; which are indicative of the physiological changes resulting from increased water supply. Plant growth rates, estimated from the variation of either shoot basal diameter or maximum height, were highly correlated with total biomass. Only LMA and δ(13)C were higher in irrigated than in control plants, but we found no further differences among irrigation treatments. This threshold response suggests an on-off strategy fitted to exploit pulses of fog water, which are always limited in magnitude in comparison with natural rain. The absence of a differential response to increased water supply is in agreement with the low phenotypic plasticity expected in plants from very stressful environments. Our results have practical implications for reforestation projects, since irrigating with 20 mm per month is sufficient to achieve the full growth capacity of this species.

Forest restoration in a fog oasis: evidence indicates need for cultural awareness in constructing the reference.

BACKGROUND: In the Peruvian Coastal Desert, an archipelago of fog oases, locally called lomas, are centers of biodiversity and of past human activity. Fog interception by a tree canopy, dominated by the legume tree tara (Caesalpinia spinosa), enables the occurrence in the Atiquipa lomas (southern Peru) of an environmental island with a diverse flora and high productivity. Although this forest provides essential services to the local population, it has suffered 90% anthropogenic reduction in area. Restoration efforts are now getting under way, including discussion as to the most appropriate reference ecosystem to use. METHODOLOGY/PRINCIPAL FINDINGS: Genetic diversity of tara was studied in the Atiquipa population and over a wide geographical and ecological range. Neither exclusive plastid haplotypes to loma formations nor clear geographical structuring of the genetic diversity was found. Photosynthetic performance and growth of seedlings naturally recruited in remnant patches of loma forest were compared with those of seedlings recruited or planted in the adjacent deforested area. Despite the greater water and nitrogen availability under tree canopy, growth of forest seedlings did not differ from that of those recruited into the deforested area, and was lower than that of planted seedlings. Tara seedlings exhibited tight stomatal control of photosynthesis, and a structural photoprotection by leaflet closure. These drought-avoiding mechanisms did not optimize seedling performance under the conditions produced by forest interception of fog moisture. CONCLUSIONS/SIGNIFICANCE: Both weak geographic partitioning of genetic variation and lack of physiological specialization of seedlings to the forest water regime strongly suggest that tara was introduced to lomas by humans. Therefore, the most diverse fragment of lomas is the result of landscape management and resource use by pre-Columbian cultures. We argue that an appropriate reference ecosystem for ecological restoration of lomas should include sustainable agroforestry practices that emulate the outcomes of ancient uses.

Alternative methods for sampling and preservation of photosynthetic pigments and tocopherols in plant material from remote locations.

Current methods for the study of pigments involve freezing in liquid nitrogen and storage at -80 degrees C or lyophilization until HPLC analysis. These requirements greatly restrict ecophysiological research in remote areas where such resources are hardly available. We aimed to overcome such limitations by developing several techniques not requiring freezing or lyophilization. Two species with contrasting foliar characteristics (Olea europaea and Taraxacum officinale) were chosen. Seven preservation methods were designed, optimized and tested in a field trial. These protocols were compared with a control immediately frozen after collection. Pigments and tocopherols were analysed by HPLC. Main artefacts were chlorophyll epimerization or phaeophytinization, carotenoid isomerization, altered de-epoxidation index and tocopherol degradation. Among all methods, sample desiccation in silica gel provides robust samples (pigment composition was unaffected by storage time or temperature) and almost unaltered pigment profiles, except for a shift in epoxidation state. Although liquid nitrogen freezing and subsequent lyophilization or freezer storage were preferred, when these facilities are either not available or not suitable for long-distance transport, desiccation with silica gel, passive extraction in acetone and/or storage of fresh samples in water vapour saturated atmospheres enable a complete pigment characterization. Silica gel is advisable for long-term sample conservation.

Dark induction of the photoprotective xanthophyll cycle in response to dehydration.

Some plants tolerate tissue dehydration. Dehydration conditions suppress photosynthesis, exacerbating photooxidative stress. In this study, fern samples were collected from the field, desiccated in darkness, and subsequently re-watered. During dark dehydration, zeaxanthin (Z) was formed and maximal photochemical efficiency of PS II was strongly reduced. Rehydration in the dark reversed these effects. Violaxanthin de-epoxidase was responsible for the dark formation of Z as illustrated by its complete inhibition by DTT. Nonetheless, its activity was not affected by nigericin, indicating that Z formation in the dark could be a process independent of the transmembrane pH-gradient into the thylakoids. Synthesis de novo of Z was rejected after blocking carotenogenesis with norfluorazon. Dark formation of Z was also observed in dehydrating leaves of desiccation-intolerant plants, which seems to indicate that this is a phenomenon scattered among different taxa within the plant kingdom. Plants may trigger this mechanism during dehydration, for chlorophyll protection during desiccation, and for faster acclimation when rehydrating conditions return. Violaxanthin de-epoxidation to form Z is typically a light-dependent process, but the formation induced solely by dehydration might represent an anticipatory mechanism for preventing early morning photodamage in desiccation-tolerant plants such as the fern Ceterach officinarum.

Phenotypic plasticity and integration across the canopy of Olea europaea subsp. guanchica (Oleaceae) in populations with different wind exposures.

Woody plants, as sessile and long-lived organisms, are expected to have effective mechanisms for dealing with recurrent environmental stresses. In the present study, we hypothesized that phenotypic plasticity (the ability to express alternative phenotypes) and integration (covariation among functionally related traits) are elicited in plants under stressful wind speed conditions. We investigated the within-crown variation of nine vegetative traits of a tree species (Olea europaea subsp. guanchica) in six populations that represented a gradient of wind speed exposures. Wind-exposed twigs in outer-canopy layers had smaller leaves; thinner, lighter, and shorter internodes; and a larger internode cross-sectional area to leaf area ratio. Comparison between field and greenhouse trials revealed that field differences among populations were mediated by phenotypic plasticity. Outer-canopy twigs expressed plastic responses in populations exposed to high wind speeds, whereas inner-canopy twigs displayed high phenotypic convergence among populations. In addition, phenotypic integration increased with wind exposure (outer canopy > inner canopy > greenhouse) and was consequently affected by canopy openness. We conclude that exposure to wind above a certain speed threshold in this woody species elicits a plastic response that is associated with increased integration among traits and involves mechanical and hydraulic rearrangements in more exposed parts of the trees.

Field patterns of leaf plasticity in adults of the long-lived evergreen Quercus coccifera.

BACKGROUND AND AIMS: Quercus coccifera, as a long-lived sprouter, responds plastically to environmental variation. In this study, the role of foliar plasticity as a mechanism of habitat selection and modification within the canopy and across contrasted habitats was characterized. An examination was made of the differential contribution of inner and outer canopy layers to the crown plasticity expressed in the field by adult individuals and its dependence on environmental and genetic factors. METHODS: Within-crown variation in eight foliar traits was examined in nine populations dominated by Q. coccifera. The difference between mean trait values at the inner and outer canopy layers was used as a proxy for crown plasticity to light. Correlations between geographic distances, environmental differences (climatic and edaphic) and phenotypic divergence (means and plasticities) were assessed by partial Mantel tests. A subset of field measurements was compared with data from a previous common garden experiment. KEY RESULTS: Phenotypic adjustment of sun leaves contributed significantly to the field variation in crown plasticity. Plasticity in leaf angle, lobation, xanthophyll cycle pigments and beta-carotene content was expressed in sun and shade leaves concurrently and in opposite directions. Phenotypic plasticity was more strongly correlated with environmental variation than mean trait values. Populations of taller plants with larger, thinner (higher specific leaf area) and less spiny leaves exhibited greater plasticity. In these populations, the midday light environment was more uniform at the inner than at the outer canopy layers. Field and common garden data ranked populations in the same order of plasticity. CONCLUSIONS: The expression of leaf plasticity resulted in a phenotypic differentiation that suggests a mechanism of habitat selection through division of labour across canopy layers. Signs of plasticity-mediated habitat modification were found only in the most plastic populations. Intracanopy plasticity was sensitive to environmental variation but also exhibited a strong genetic component.

Extensive gene flow blurs phylogeographic but not phylogenetic signal in Olea europaea L.

Genetic structure and evolutionary patterns of the wild olive tree (Olea europaea L.) were investigated with AFLP fingerprinting data at three geographic levels: (a) phylogenetic relationships of the six currently recognized subspecies in Eurasia and Africa; (b) lineage identification in subsp. europaea of the Mediterranean basin; and (c) phylogeography in the western Mediterranean. Two statistical approaches (Bayesian inference and analysis of molecular variance) were used to analyse the AFLP fingerprints. To determine the congruency and transferability of results across studies previous RAPD and ISSR data were analysed in a similar manner. Comparisons proved that qualitative results were mostly congruent but quantitative values differed, depending on the method of analysis. Neighbour-Joining analysis of AFLP phenotypes supported current classification of subspecies. At a Mediterranean scale no clear cut phylogeographic pattern was recovered, likely due to extensive gene flow between populations of subsp. europaea. Gene flow estimates calculated with conventional F-statistics showed that reproductive barriers separated neither populations nor lineages of O. europaea. Genetic divergence between eastern and western parts of the Mediterranean basin was observed only when geographical and population information were incorporated into the analyses through hierarchical analysis of molecular variance (AMOVA). Within the western Mediterranean, the highest genetic diversity was found in two regions: on both sides of the Strait of Gibraltar and in the Balearic archipelago. Additionally, long-lasting isolation of the northern-most populations of the Iberian Peninsula appeared to be responsible for a significant divergence.

Mechanisms underlying the amelioration of O3-induced damage by elevated atmospheric concentrations of CO2.

There is growing evidence that rising atmospheric CO2 concentrations will reduce or prevent reductions in the growth and productivity of C3 crops attributable to ozone (O3) pollution. In this study, the role of pollutant exclusion in mediating this response was investigated through growth chamber-based investigations on leaves 4 and 7 of spring wheat (Triticum aestivum cv. Hanno). In the core experiments, plants were raised at two atmospheric CO2 concentrations (ambient [350 micro l l(-1)] or elevated CO2 [700 micro l l(-1)] under two O3 regimes (charcoal/Purafil-filtered air [<5 nl l(-1) O3] or ozone-enriched air [75 nl l(-1) 7 h d(-1)]). A subsequent experiment used an additional O3 treatment where the goal was to achieve equivalent daily O3 uptake over the life-span of leaves 4 and 7 under ambient and CO2-enriched conditions, through daily adjustment of exposures based on measured shifts in stomatal conductance. Plant growth and net CO2 assimilation were stimulated by CO2-enrichment and reduced by exposure to O3. However, the impacts of O3 decreased with plant age (i.e. leaf 7 was more resistant to O3 injury than leaf 4); a finding consistent with ontogenic shifts in the tolerance of plant tissue and/or acclimation to O3-induced oxidative stress. In the combined treatment, elevated CO2 protected against the adverse effects of O3 and reduced cumulative O3 uptake (calculated from measurements of stomatal conductance) by c. 10% and 35% over the life-span of leaves 4 and 7, respectively. Analysis of the relationship between O3 uptake and the decline in the maximum in vivo rate of Rubisco carboxylation (Vcmax) revealed the protection afforded by CO2-enrichment to be due, to a large extent, to the exclusion of the pollutant from the leaf interior (as a consequence of the decline in stomatal conductance triggered by CO2-enrichment), but there was evidence (especially from flux-response relationships constructed for leaf 4) that CO2-enrichment resulted in additional effects that alleviated the impacts of ozone-induced oxidative stress on photosynthesis.

Plasticity, instability and canalization: is the phenotypic variation in seedlings of sclerophyll oaks consistent with the environmental unpredictability of Mediterranean ecosystems?

• Evergreen oaks from the Mediterranean basin exhibit a conservative resource-use strategy based on a reduced expression of phenotypic variation (i.e. canalization). We hypothesized that genetic variation across closely related species is more canalized than the response to environmental variation. • Seedlings of Quercus ilex and Q. coccifera, two important oak species from the Mediterranean basin that belong to the same subgenus and section, were grown in contrasted light and nutrient environments following a factorial design. Phenotypic variation was explored in a total of 75 variables including photosynthetic capacity, nutrient allocation, allometric relationships and crown architecture. • Path analysis showed that phenotypic variation was not significantly affected by differences between species but by those between and within environments, which are argued to be primarily linked to phenotypic plasticity and developmental instability, respectively. This finding is interpreted as evidence of genetic canalization across species. • The similar importance of plasticity and instability as sources of phenotypic variation and the high degree of genetic canalization are consistent with the expected role of the environmental unpredictability of Mediterranean ecosystems in shaping the developmental patterns of these two species.

Low leaf-level response to light and nutrients in Mediterranean evergreen oaks: a conservative resource-use strategy?

We have explored leaf-level plastic response to light and nutrients of Quercus ilex and Q. coccifera, two closely related Mediterranean evergreen sclerophylls, in a factorial experiment with seedlings. Leaf phenotypic plasticity, assessed by a relative index (PI = (maximum value - minimum)/maximum) in combination with the significance of the difference among means, was studied in 37 morphological and physiological variables. Light had significant effects on most variables relating to photosynthetic pigments, chlorophyll fluorescence and gas exchange, whereas nutrient treatment had a significant effect in only 10% of the variables. Chlorophyll content was higher in the shade whereas carotenoid content and nonphotochemical quenching increased with light. Nutrient limitations increased the xanthophyll-cycle pool but only at high light intensities, and the same interaction between light and nutrients was observed for lutein. Predawn photochemical efficiency of PSII was not affected by either light or nutrients, although midday photochemical efficiency of PSII was lower at high light intensities. Photosynthetic light compensation point and dark respiration on an area basis decreased with light, but photosynthetic capacity on a dry mass basis and photochemical quenching were higher in low light, which translated into a higher nitrogen use efficiency in the shade. We expected Q. ilex, the species of the widest ecological distribution, to be more plastic than Q. coccifera, but differences were minor: Q. ilex exhibited a significant response to light in 13% more of the variables than Q. coccifera, but mean PI was very similar in the two species. Both species tolerated full sunlight and moderate shade, but exhibited a reduced capacity to enhance photosynthetic utilization of high irradiance. When compared with evergreen shrubs from the tropical rainforest, leaf responsiveness of the two evergreen oaks was low. We suggest that the low leaf-level responsiveness found here is part of a conservative resource use strategy, which seems to be adaptive for evergreen woody plants in Mediterranean-type ecosystems.

Potential effects of rising tropospheric concentrations of CO2 and O3 on green-algal lichens.

Pormelia sulcata Taylor was used as a model to examine the effects of elevated CO2 and/or O3 on green algal lichens. Thalli were exposed for 30 d in duplicate controlled-environment chambers to two atmospheric concentrations of CO2 ('ambient' [350µmol mol-1 ] and 'elevated' [700µmol mol-1 ] 24 h d-1 ) and two O3 regimes ('non-polluted' air [CF, < 5 nmol mol-1 ] and 'polluted' air [15 nmol mol-1 overnight rising to a midday maximum of 75 nmol mol-1 ]), in a factorial design. Elevated CO2 , or elevated O3 depressed the light saturated rate of CO2 , assimilation Asat ) measured at ambient CO2 , by 30% and 18%, respectively. However, despite this effect ultrastructure) studies revealed increased lipid storage in cells of the photobiont in response to CO2 -enrichment. Simultaneous exposure to elevated O3 reduced CO2 -induced lipid accumulation and reduced Asat in an additive manner. Gold-antibody labelling revealed that the decline in photosynthetic capacity induced by elevated CO2 and/or O3 was accompanied by a parallel decrease in the concentration of Rubiscoa in the algal pyrenoid (r= 0.93). Interestingly, differences in the amount of Rubisco protein were not correlated with changes in pyrenoid volume. Measurements of in vivo chlorophyll-fluorescence induction kinetics showed that the decline in Asat induced by elevated CO2 , and/or O2 , was not associated with significant changes in the photochemical efficiency of photosystem (PS) II. Although the experimental conditions inevitably imposed some stress on the thalli, revealed as a significant decline in the efficiency of PS II photochemistry, and enhanced starch accumulation in the photobiont over the fornication period, the study shows that the green-algal lichen symbiosis might be influenced by future changes in atmospheric composition. Photosynthetic capacity, measured at ambient CO2 , was found to be reduced after a controlled 30 d exposure to elevated CO2 , and/or O3 and this effect was associated with a parallel decline in the amount of Rubisco in the pyrenoid of algal chloroplasts.