Influence of increased nutrient availability on biogenic volatile organic compound (BVOC) emissions and leaf anatomy of subarctic dwarf shrubs under climate warming and increased cloudiness.

BACKGROUND AND AIMS: Climate change is subjecting subarctic ecosystems to elevated temperature, increased nutrient availability and reduced light availability (due to increasing cloud cover). This may affect subarctic vegetation by altering the emissions of biogenic volatile organic compounds (BVOCs) and leaf anatomy. We investigated the effects of increased nutrient availability on BVOC emissions and leaf anatomy of three subarctic dwarf shrub species, Empetrum hermaphroditum, Cassiope tetragona and Betula nana, and if increased nutrient availability modifies the responses to warming and shading. METHODS: Measurements of BVOCs were performed in situ in long-term field experiments in the Subarctic using a dynamic enclosure system and collection of BVOCs into adsorbent cartridges analysed by gas chromatography-mass spectrometry. Leaf anatomy was studied using light microscopy and scanning electron microscopy. KEY RESULTS: Increased nutrient availability increased monoterpene emission rates and altered the emission profile of B. nana, and increased sesquiterpene and oxygenated monoterpene emissions of C. tetragona. Increased nutrient availability increased leaf tissue thicknesses of B. nana and C. tetragona, while it caused thinner epidermis and the highest fraction of functional (intact) glandular trichomes for E. hermaphroditum. Increased nutrient availability and warming synergistically increased mesophyll intercellular space of B. nana and glandular trichome density of C. tetragona, while treatments combining increased nutrient availability and shading had an opposite effect in C. tetragona. CONCLUSIONS: Increased nutrient availability may enhance the protection capacity against biotic and abiotic stresses (especially heat and drought) in subarctic shrubs under future warming conditions as opposed to increased cloudiness, which could lead to decreased resistance. The study emphasizes the importance of changes in nutrient availability in the Subarctic, which can interact with climate warming and increased cloudiness effects.

Alpine dwarf shrubs show high proportions of nonfunctional xylem: Visualization and quantification of species-specific patterns.

Xylem conductive capacity is a key determinant of plant hydraulic function and intimately linked to photosynthesis and productivity, but can be impeded by temporary or permanent conduit dysfunctions. Here we show that persistent xylem dysfunctions in unstressed plants are frequent in Alpine dwarf shrubs and occur in various but species-specific cross-sectional patterns. Combined synchrotron micro-computed tomography (micro-CT) imaging, xylem staining, and flow measurements in saturated samples of six widespread Ericaceae species evidence a high proportion (19%-50%) of hydraulically nonfunctional xylem areas in the absence of drought stress, with regular distribution of dysfunctions between or within growth rings. Dysfunctions were only partly reversible and reduced the specific hydraulic conductivity to 1.38 to 3.57 ×10-4 m2 s-1 MPa-1 . Decommission of inner growth rings was clearly related to stem age and a higher vulnerability to cavitation of older rings, while the high proportion of nonfunctional conduits in each annual ring needs further investigations. The lower the xylem fraction contributing to the transport function, the higher was the hydraulic efficiency of conducting xylem areas. Improved understanding of the functional lifespan of xylem elements and the prevalence and nature of dysfunctions is critical to correctly assess structure-function relationships and whole-plant hydraulic strategies.

Strategies in a metallophyte species to cope with manganese excess.

The effect of exposure to high Mn concentration was studied in a metallophyte species, Erica andevalensis, using hydroponic cultures with a range of Mn concentrations (0.06, 100, 300, 500, and 700 mg L-1). At harvest, biomass production, element uptake, and biochemical indicators of metal stress (leaf pigments, organic acids, amino acids, phenols, and activities of catalase, peroxidase, superoxide dismutase) were determined in leaves and roots. Increasing Mn concentrations led to a decrease in biomass accumulation, and tip leaves chlorosis was the only toxicity symptom detected. In a similar way, photosynthetic pigments (chlorophylls a and b, and carotenoids) were affected by high Mn levels. Among organic acids, malate and oxalate contents in roots showed a significant increase at the highest Mn concentration, while in leaves, Mn led to an increasing trend in citrate and malate contents. An increase of Mn also induced an increase in superoxide dismutase activity in roots and catalase activity in leaves. As well, significant changes in free amino acids were induced by Mn concentrations higher than 300 mg L-1, especially in roots. No significant changes in phenolic compounds were observed in the leaves, but root phenolics were significantly increased by increasing Mn concentrations in treatments. When Fe supply was increased 10 and 20 times (7-14 mg Fe L-1 as Fe-EDDHA) in the nutrient solutions at the highest Mn concentration (700 mg Mn L-1), it led to significant increases in photosynthetic pigments and biomass accumulation. Manganese was mostly accumulated in the roots, and the species was essentially a Mn excluder. However, considering the high leaf Mn concentration recorded without toxicity symptoms, E. andevalensis might be rated as a Mn-tolerant species.

Post-pollination barriers enable coexistence of pollinator-sharing ornithophilous Erica species.

Some evolutionary radiations produce a number of closely-related species that continue to coexist. In such plant systems, when pre-pollination barriers are weak, relatively strong post-pollination reproductive barriers are required to maintain species boundaries. Even when post-pollination barriers are in place, however, reproductive interference and pollinator dependence may strengthen selection for pre-pollination barriers. We assessed whether coexistence of species from the unusually speciose Erica genus in the fynbos biome, South Africa, is enabled through pre-pollination or post-pollination barriers. We also tested for reproductive interference and pollinator dependence. We investigated this in natural populations of three bird-pollinated Erica species (Erica plukenetii, E. curviflora and E. coccinea), which form part of a large guild of congeneric species that co-flower and share a single pollinator species (Orange-breasted Sunbird Anthobaphes violacea). At least two of the three pre-pollination barriers assessed (distribution ranges, flowering phenology and flower morphology) were weak in each species pair. Hand-pollination experiments revealed that seed set from heterospecific pollination (average 8%) was significantly lower than seed set from outcross pollination (average 50%), supporting the hypothesis that species boundaries are maintained through post-pollination barriers. Reproductive interference, assessed in one population by applying outcross pollen three hours after applying heterospecific pollen, significantly reduced seed set compared to outcross pollen alone. This may drive selection for traits that enhance pre-pollination barriers, particularly given that two of the three species were self-sterile, and therefore pollinator dependent. This study suggests that post-pollination reproductive barriers could facilitate the coexistence of congeneric species, in a recent radiation with weak pre-pollination reproductive barriers.

Phytostabilization potential of Erica australis L. and Nerium oleander L.: a comparative study in the Riotinto mining area (SW Spain).

Phytostabilization is a green, cost-effective technique for mine rehabilitation and ecological restoration. In this study, the phytostabilization capacity of Erica australis L. and Nerium oleander L. was assessed in the climatic and geochemical context of the Riotinto mining district, southwestern Spain, where both plant species colonize harsh substrates of mine wastes and contaminated river banks. In addition to tolerating extreme acidic conditions (up to pH 3.36 for E. australis), both species were found to grow on substrates very poor in bioavailable nutrients (e.g., N and P) and highly enriched with potentially phytotoxic elements (e.g., Cu, Cd, Pb, S). The selective root absorption of essential elements and the sequestration of potentially toxic elements in the root cortex are the main adaptations that allow the studied species to cope in very limiting edaphic environments. Being capable of a tight elemental homeostatic control and tolerating extreme acidic conditions, E. australis is the best candidate for use in phytostabilization programs, ideally to promote early stages of colonization, improve physical and chemical conditions of substrates and favor the establishing of less tolerant species, such as N. oleander.

Simulated projections of boreal forest peatland ecosystem productivity are sensitive to observed seasonality in leaf physiology†.

We quantified seasonal CO2 assimilation capacities for seven dominant vascular species in a wet boreal forest peatland then applied data to a land surface model parametrized to the site (ELM-SPRUCE) to test if seasonality in photosynthetic parameters results in differences in simulated plant responses to elevated CO2 and temperature. We collected seasonal leaf-level gas exchange, nutrient content and stand allometric data from the field-layer community (i.e., Maianthemum trifolium (L.) Sloboda), understory shrubs (Rhododendron groenlandicum (Oeder) Kron and Judd, Chamaedaphne calyculata (L.) Moench., Kalmia polifolia Wangenh. and Vaccinium angustifolium Alton.) and overstory trees (Picea mariana (Mill.) B.S.P. and Larix laricina (Du Roi) K. Koch). We found significant interspecific seasonal differences in specific leaf area, nitrogen content (by area; Na) and photosynthetic parameters (i.e., maximum rates of Rubisco carboxylation (Vcmax25°C), electron transport (Jmax25°C) and dark respiration (Rd25°C)), but minimal correlation between foliar Na and Vcmax25°C, Jmax25°C or Rd25°C, which illustrates that nitrogen alone is not a good correlate for physiological processes such as Rubisco activity that can change seasonally in this system. ELM-SPRUCE was sensitive to the introduction of observed interspecific seasonality in Vcmax25°C, Jmax25°C and Rd25°C, leading to simulated enhancement of net primary production (NPP) using seasonally dynamic parameters as compared with use of static parameters. This pattern was particularly pronounced under simulations with higher temperature and elevated CO2, suggesting a key hypothesis to address with future empirical or observational studies as climate changes. Inclusion of species-specific seasonal photosynthetic parameters should improve estimates of boreal ecosystem-level NPP, especially if impacts of seasonal physiological ontogeny can be separated from seasonal thermal acclimation.

Social wasps, crickets and cockroaches contribute to pollination of the holoparasitic plant Mitrastemon yamamotoi (Mitrastemonaceae) in southern Japan.

Mitrastemon yamamotoi is completely embedded within the tissues of its hosts, except during the reproductive stage, when aboveground parts emerge from host tissues. Its highly modified appearance has attracted attention of many botanists, but very little is known about the reproductive system. Floral visitors to M. yamamotoi were observed in southern Japan. Pollination experiments were conducted to determine the plant's self-compatibility and pollen limitation, as well as the contribution of diurnal and nocturnal visitors to fruit set and outcrossing. Mitrastemon yamamotoi is mainly pollinated by social wasps, but previously unnoticed pollinators (i.e. crickets and cockroaches) are also important, based on visitation frequency and pollen loads. Results of the pollination experiments suggest that nocturnal visitors, such as crickets and cockroaches, contribute to geitonogamous pollination, whereas diurnal visitors, such as social wasps, facilitate outcrossing. The unexpected pollinator assemblage of M. yamamotoi might be influenced by multiple factors, including the highly modified flowers that are produced close to the ground in dark understorey environments, the species' winter-flowering habit and the location of the study site (i.e. near the northern limit of the species' range). Considering that M. yamamotoi occurs widely in subtropical and tropical forests in Asia, additional studies are needed to assess pollinator assemblages of M. yamamotoi at other locations.

Phenotypic plasticity of floral volatiles in response to increasing drought stress.

BACKGROUND AND AIMS: Flowers emit a wide range of volatile compounds which can be critically important to interactions with pollinators or herbivores. Yet most studies of how the environment influences plant volatiles focus on leaf emissions, with little known about abiotic sources of variation in floral volatiles. Understanding phenotypic plasticity in floral volatile emissions has become increasingly important with globally increasing temperatures and changes in drought frequency and severity. Here quantitative relationships of floral volatile emissions to soil water content were analysed. METHODS: Plants of the sub-alpine herb Ipomopsis aggregata and hybrids with its closest congener were subjected to a progressive dry down, mimicking the range of soil moistures experienced in the field. Floral volatiles and leaf gas exchange were measured at four time points during the drought. KEY RESULTS: As the soil dried, floral volatile emissions increased overall and changed in composition, from more 1,3-octadiene and benzyl alcohol to higher representation of some terpenes. Emissions of individual compounds were not linearly related to volumetric water content in the soil. The dominant compound, the monoterpene α-pinene, made up the highest percentage of the scent mixture when soil moisture was intermediate. In contrast, emission of the sesquiterpene (E,E)-α-farnesene accelerated as the drought became more intense. Changes in floral volatiles did not track the time course of changes in photosynthetic rate or stomatal conductance. CONCLUSIONS: This study shows responses of specific floral volatile organic compounds to soil moisture. The non-linear responses furthermore suggest that extreme droughts may have impacts that are not predictable from milder droughts. Floral volatiles are likely to change seasonally with early summer droughts in the Rocky Mountains, as well as over years as snowmelt becomes progressively earlier. Changes in water availability may have impacts on plant-animal interactions that are mediated through non-linear changes in floral volatiles.

Dispensing Pollen via Catapult: Explosive Pollen Release in Mountain Laurel (Kalmia latifolia).

The astonishing amount of floral diversity has inspired countless assumptions about the function of diverse forms and their adaptive significance, yet many of these hypothesized functions are untested. We investigated an often-repeated adaptive hypothesis about how an extreme floral form functions. In this study, we conducted four investigations to understand the adaptive function of explosive pollination in Kalmia latifolia, the mountain laurel. We first performed a kinematic analysis of anther movement. Second, we constructed a heat map of pollen trajectories in three-dimensional space. Third, we conducted field observations of pollinators and their behaviors while visiting K. latifolia. Finally, we conducted a pollination experiment to investigate the importance of pollinators for fertilization success. Our results suggest that insect visitation dramatically improves fertilization success and that bees are the primary pollinators that trigger explosive pollen release.

Xylogenesis reveals the genesis and ecological signal of IADFs in Pinus pinea L. and Arbutus unedo L.

Background and Aims: Mediterranean trees have patterns of cambial activity with one or more pauses per year, leading to intra-annual density fluctuations (IADFs) in tree rings. We analysed xylogenesis (January 2015-January 2016) in Pinus pinea L. and Arbutus unedo L., co-occurring at a site on Mt. Vesuvius (southern Italy), to identify the cambial productivity and timing of IADF formation. Methods: Dendrochronological methods and quantitative wood anatomy were applied and enabled IADF identification and classification. Key Results: We showed that cambium in P. pinea was productive throughout the calendar year. From January to March 2015, post-cambial (enlarging) earlywood-like tracheids were observed, which were similar to transition tracheids. The beginning of the tree ring was therefore not marked by a sharp boundary between latewood of the previous year and the new xylem produced. True earlywood tracheids were formed in April. L-IADFs were formed in autumn, with earlywood-like cells in latewood. In A. unedo, a double pause in cell production was observed, in summer and winter, leading to L-IADFs in autumn as well. Moreover, the formation of more than one IADF was observed in A. unedo. Conclusions: Despite having completely different wood formation models and different life strategies, the production of earlywood, latewood and IADF cells was strongly controlled by climatic factors in the two species. Such cambial production patterns need to be taken into account in dendroecological studies to interpret climatic signals in wood from Mediterranean trees.

Phyto-toponyms of Arbutus unedo L. and their distribution in Sardinia (Italy).

The study shows the results of an inventory of place names connected to Arbutus unedo L., a Mediterranean species, widespread throughout Sardinia. The main aim was to compare the past distribution of place names, referring to the strawberry tree, to the current distribution of the species on the island. In addition, we investigated the meaning and the diversity of these local place names in the various communities. The result was a collection of 432 phyto-toponyms. 248 of them were used for an analysis of their distribution in the habitats, indicated on the Map of the Nature System in Sardinia, defined on the basis of the current vegetation typology. The persistence of the species in the various habitats was either confirmed or negated with in site investigations and interviews. 47.5% of municipalities have place names related to the strawberry tree. Of the 248 phyto-toponyms, 127 fall in the habitats where the species currently persists proving a correspondence between their regional distribution and the current distribution of the species. The remaining 121 phyto-toponyms fall in habitats where the strawberry tree is currently absent. Most of them are found in man-made habitats where man has transformed the forest cover which previously included the strawberry tree. This study also contributes to promoting and conserving the linguistic heritage of local communities.

On the brink: the highly reduced plastomes of nonphotosynthetic Ericaceae.

Ericaceae (the heather family) is a large and diverse group of plants that forms elaborate symbiotic relationships with mycorrhizal fungi, and includes several nonphotosynthetic lineages. Using an extensive sample of fully mycoheterotrophic (MH) species, we explored inter- and intraspecific variation as well as selective constraints acting on the plastomes of these unusual plants. The plastomes of seven MH genera were analysed in a phylogenetic context with two geographically disparate individuals sequenced for Allotropa, Monotropa, and Pityopus. The plastomes of nonphotosynthetic Ericaceae are highly reduced in size (c. 33-41 kbp) and content, having lost all photosynthesis-related genes, and are reduced to encoding housekeeping genes as well as a protease subunit (clpP)-like and acetyl-CoA carboxylase subunit D (accD)-like open reading frames. Despite an increase in the rate of their nucleotide substitutions, the remaining protein-coding genes are typically under purifying selection in full MHs. We also identified ribosomal proteins under relaxed or neutral selection. These plastomes also exhibit striking structural rearrangements. Intraspecific variation within MH Ericaceae ranges from a few differences (Allotropa) to extensive population divergences (Monotropa, Hypopitys), which indicates that cryptic speciation may be occurring in several lineages. The pattern of gene loss within fully MH Ericaceae plastomes suggests an advanced state of degradation.

Corema album: unbiased dioecy in a competitive environment.

Corema album is a dioecious coastal shrub. Dioecious plants growing in these resource-limited habitats may present spatial segregation of the sexes (SSS) or demographic biases because of the different reproductive effort between sexes. In these environments facilitation is a more common interaction between plants than competition. To assess factors determining the distribution of C. album male and female plants, we investigated the influence of habitat type (sand dunes and coastal woodlands), assessed the occurrence of SSS or demographic biases and also a possible role of these shrubs as nurse plants. We selected three C. album populations with the two habitat types. All C. album individuals were sexed, mapped and measured in three plots (20 m × 20 m) per population/habitat type. Presence and abundance of all plant species were recorded under five female and five male C. album plants as well as in equivalent open ground area in each of 15 plots. According to Ripley's K function result, C. album did not display SSS. Generalised linear mixed models (GLMM) show that differences in plant size were not related to plant sex. Plant inventory correspondence analysis showed that species composition and abundance were influenced by habitat type, population and the presence of a C. album individual, but not by its sex. GLMM indicated a detrimental effect of C. album on the co-occurring plants. Our results show that sexual dimorphism has allowed C. album to adapt to the environment avoiding SSS or significant demographic bias, suggesting a positive outlook for its conservation.

Identification and expression analysis of chitinase genes in parasitic plant Monotropa hypopitys.

Genes encoding six chitinases, five of which belong to classes I (MhCHI3 and MhCHI4), IV (MhCHI1), V (MhCHI5), and VII (MhCHI2), were identified in the transcriptome of the parasitic mixoheterotrophic plant Monotropa hypopitys. The transcription level of MhCHI5 and MhCHI1 was low; however, in the leaves (bracts) and roots it was higher than in flowers. MhCHI4 transcripts were detected primarily in the flowers and were almost absent in the roots, whereas the expression level of MhCHI3 was relatively high in all organs but maximum in the leaves (bracts).

Specificity of fungal associations of Pyroleae and Monotropa hypopitys during germination and seedling development.

Mycoheterotrophic plants obtain organic carbon from associated mycorrhizal fungi, fully or partially. Angiosperms with this form of nutrition possess exceptionally small 'dust seeds' which after germination develop 'seedlings' that remain subterranean for several years, fully dependent on fungi for supply of carbon. Mycoheterotrophs which as adults have photosynthesis thus develop from full to partial mycoheterotrophy, or autotrophy, during ontogeny. Mycoheterotrophic plants may represent a gradient of variation in a parasitism-mutualism continuum, both among and within species. Previous studies on plant-fungal associations in mycoheterotrophs have focused on either germination or the adult life stages of the plant. Much less is known about the fungal associations during development of the subterranean seedlings. We investigated germination and seedling development and the diversity of fungi associated with germinating seeds and subterranean seedlings (juveniles) in five Monotropoideae (Ericaceae) species, the full mycoheterotroph Monotropa hypopitys and the putatively partial mycoheterotrophs Pyrola chlorantha, P. rotundifolia, Moneses uniflora and Chimaphila umbellata. Seedlings retrieved from seed sowing experiments in the field were used to examine diversity of fungal associates, using pyrosequencing analysis of ITS2 region for fungal identification. The investigated species varied with regard to germination, seedling development and diversity of associated fungi during juvenile ontogeny. Results suggest that fungal host specificity increases during juvenile ontogeny, most pronounced in the fully mycoheterotrophic species, but a narrowing of fungal associates was found also in two partially mycoheterotrophic species. We suggest that variation in specificity of associated fungi during seedling ontogeny in mycoheterotrophs represents ongoing evolution along a parasitism-mutualism continuum.

The light response of mesophyll conductance is controlled by structure across leaf profiles.

Mesophyll conductance to CO2 (gm ) may respond to light either through regulated dynamic mechanisms or due to anatomical and structural factors. At low light, some layers of cells in the leaf cross-section approach photocompensation and contribute minimally to bulk leaf photosynthesis and little to whole leaf gm (gm,leaf ). Thus, the bulk gm,leaf will appear to respond to light despite being based upon cells having an anatomically fixed mesophyll conductance. Such behaviour was observed in species with contrasting leaf structure using the variable J or stable isotope method of measuring gm,leaf . A species with bifacial structure, Arbutus × 'Marina', and an isobilateral species, Triticum durum L., had contrasting responses of gm,leaf upon varying adaxial or abaxial illumination. Anatomical observations, when coupled with the proposed model of gm,leaf to photosynthetic photon flux density (PPFD) response, successfully represented the observed gas exchange data. The theoretical and observed evidence that gm,leaf apparently responds to light has large implications for how gm,leaf values are interpreted, particularly limitation analyses, and indicates the importance of measuring gm under full light saturation. Responses of gm,leaf to the environment should be treated as an emergent property of a distributed 3D structure, and not solely a leaf area-based phenomenon.

Leaf anatomy, BVOC emission and CO2 exchange of arctic plants following snow addition and summer warming.

BACKGROUND AND AIMS: Climate change in the Arctic is projected to increase temperature, precipitation and snowfall. This may alter leaf anatomy and gas exchange either directly or indirectly. Our aim was to assess whether increased snow depth and warming modify leaf anatomy and affect biogenic volatile organic compound (BVOC) emissions and CO2 exchange of the widespread arctic shrubs Betula nana and Empetrum nigrum ssp. hermaphroditum METHODS: Measurements were conducted in a full-factorial field experiment in Central West Greenland, with passive summer warming by open-top chambers and snow addition using snow fences. Leaf anatomy was assessed using light microscopy and scanning electron microscopy. BVOC emissions were measured using a dynamic enclosure system and collection of BVOCs into adsorbent cartridges analysed by gas chromatography-mass spectrometry. Carbon dioxide exchange was measured using an infrared gas analyser. KEY RESULTS: Despite a later snowmelt and reduced photosynthesis for B. nana especially, no apparent delays in the BVOC emissions were observed in response to snow addition. Only a few effects of the treatments were seen for the BVOC emissions, with sesquiterpenes being the most responsive compound group. Snow addition affected leaf anatomy by increasing the glandular trichome density in B. nana and modifying the mesophyll of E. hermaphroditum The open-top chambers thickened the epidermis of B. nana, while increasing the glandular trichome density and reducing the palisade:spongy mesophyll ratio in E. hermaphroditum CONCLUSIONS: Leaf anatomy was modified by both treatments already after the first winter and we suggest links between leaf anatomy, CO2 exchange and BVOC emissions. While warming is likely to reduce soil moisture, melt water from a deeper snow pack alleviates water stress in the early growing season. The study emphasizes the ecological importance of changes in winter precipitation in the Arctic, which can interact with climate-warming effects.

Drought responses, phenotypic plasticity and survival of Mediterranean species in two different microclimatic sites.

Climate models predict a further drying of the Mediterranean summer. One way for plant species to persist during such climate changes is through acclimation. Here, we determine the extent to which trait plasticity in response to drought differs between species and between sites, and address the question whether there is a trade-off between drought survival and phenotypic plasticity. Throughout the summer we measured physiological traits (photosynthesis - Amax , stomatal conductance - gs , transpiration - E, leaf water potential - ψl) and structural traits (specific leaf area - SLA, leaf density - LD, leaf dry matter content - LDMC, leaf relative water content - LRWC) of leaves of eight woody species in two sites with slightly different microclimate (north- versus south-facing slopes) in southern Spain. Plant recovery and survival was estimated after the summer drought period. We found high trait variability between species. In most variables, phenotypic plasticity was lower in the drier site. Phenotypic plasticity of SLA and LDMC correlated negatively with drought survival, which suggests a trade-off between them. On the other hand, high phenotypic plasticity of SLA and LDMC was positively related to traits associated with rapid recovery and growth after the drought period. Although phenotypic plasticity is generally seen as favourable during stress conditions, here it seemed beneficial for favourable conditions. We propose that in environments with fluctuating drought periods there can be a trade-off between drought survival and growth during favourable conditions. When climate become drier, species with high drought survival but low phenotypic plasticity might be selected for.

Germination sensitivity to water stress in four shrubby species across the Mediterranean Basin.

Mediterranean shrublands are generally water-limited and fire-driven ecosystems. Seed-based post-fire regeneration may be affected by varying rainfall patterns, depending on species sensitivity to germinate under water stress. In our study, we considered the germination response to water stress in four species from several sites across the Mediterranean Basin. Seeds of species with a hard coat (Cistus monspeliensis, C. salviifolius, Cistaceae, Calicotome villosa, Fabaceae) or soft coat (Erica arborea, Ericaceae), which were exposed or not to a heat shock and smoke (fire cues), were made to germinate under water stress. Final germination percentage, germination speed and viability of seeds were recorded. Germination was modelled using hydrotime analysis and correlated to the water balance characteristics of seed provenance. Water stress was found to decrease final germination in the three hard-seeded species, as well as reduce germination speed. Moreover, an interaction between fire cues and water stress was found, whereby fire cues increased sensitivity to water stress. Seed viability after germination under water stress also declined in two hard-seeded species. Conversely, E. arborea showed little sensitivity to water stress, independent of fire cues. Germination responses varied among populations of all species, and hydrotime parameters were not correlated to site water balance, except in E. arborea when not exposed to fire cues. In conclusion, the species studied differed in germination sensitivity to water stress; furthermore, fire cues increased this sensitivity in the three hard-seeded species, but not in E. arborea. Moreover, populations within species consistently differed among themselves, but these differences could only be related to the provenance locality in E. arborea in seeds not exposed to fire cues.

Nectar foragers contribute to the pollination of buzz-pollinated plant species.

PREMISE OF THE STUDY: Pollination performance may depend on the type of floral resource (pollen or nectar) foraged by visitors. In buzz-pollinated plant species, the poricidal anthers release pollen during active pollen collection that induces flower vibrations. These buzz-pollinated species generally do not produce nectar. Nevertheless, several Ericaceae are buzz-pollinated and produce nectar. We estimated the relative effectiveness of visitors according to the type of resource collected, nectar or pollen (buzzing). METHODS: We compared the relative performance of pollen removal, transport, and deposition (effectiveness) of the main insect visitors on four ericaceous species: three buzz-pollinated species with different pore sizes, Erica tetralix, Vaccinium myrtillus, and V. vitis-idaea; and one non-buzz-pollinated species, Calluna vulgaris. KEY RESULTS: Bumblebees were the main pollinators for the three buzz-pollinated species, whereas hoverflies were the main pollinators for the non-buzz-pollinated generalist C. vulgaris. For the studied plant species, we observed no difference in pollination effectiveness among bumblebee species. Buzzing bumblebees were the most effective visitors for pollination per flower visit for the two Vaccinium species, whereas nectar foragers were the most effective visitors for pollination of E. tetralix. In the case of Vaccinium myrtillus, nectar foragers contributed the most to pollination success because they were more abundant than pollen foragers. CONCLUSIONS: We showed that consideration of the resource collected by visitors and their behavior is necessary to compare their relative performance. The combination of visitation rate and effectiveness per visit reveals that nectar foragers make a substantial contribution to pollination of the buzz-pollinated ericaceous species.