Transcriptomic analysis of flower opening response to relatively low temperatures in Osmanthus fragrans.

BACKGROUND: Sweet osmanthus (Osmanthus fragrans Lour.) is one of the top ten traditional ornamental flowers in China. The flowering time of once-flowering cultivars in O. fragrans is greatly affected by the relatively low temperature, but there are few reports on its molecular mechanism to date. A hypothesis had been raised that genes related with flower opening might be up-regulated in response to relatively low temperature in O. fragrans. Thus, our work was aimed to explore the underlying molecular mechanism of flower opening regulated by relatively low temperature in O. fragrans. RESULTS: The cell size of adaxial and abaxial petal epidermal cells and ultrastructural morphology of petal cells at different developmental stages were observed. The cell size of adaxial and abaxial petal epidermal cells increased gradually with the process of flower opening. Then the transcriptomic sequencing was employed to analyze the differentially expressed genes (DEGs) under different number of days' treatments with relatively low temperatures (19 °C) or 23 °C. Analysis of DEGs in Gene Ontology analysis showed that "metabolic process", "cellular process", "binding", "catalytic activity", "cell", "cell part", "membrane", "membrane part", "single-organism process", and "organelle" were highly enriched. In KEGG analysis, "metabolic pathways", "biosynthesis of secondary metabolites", "plant-pathogen interaction", "starch and sucrose metabolism", and "plant hormone signal transduction" were the top five pathways containing the greatest number of DEGs. The DEGs involved in cell wall metabolism, phytohormone signal transduction pathways, and eight kinds of transcription factors were analyzed in depth. CONCLUSIONS: Several unigenes involved in cell wall metabolism, phytohormone signal transduction pathway, and transcription factors with highly variable expression levels between different temperature treatments may be involved in petal cell expansion during flower opening process in response to the relatively low temperature. These results could improve our understanding of the molecular mechanism of relatively-low-temperature-regulated flower opening of O. fragrans, provide practical information for the prediction and regulation of flowering time in O. fragrans, and ultimately pave the way for genetic modification in O. fragrans.

Functional androdioecy in the ornamental shrub Osmanthus delavayi (Oleaceae).

Androdioecy is one of the rarest sexual systems among plants, characterized by males co-occurring with hermaphrodites. Osmanthus delavayi (Oleaceae), an ornamental shrub from southern China, is known to have both male and hermaphrodite individuals, but little is known regarding the breeding system of this species and whether it is functionally androdioecious, and how this potentially evolved. In this study, we explore the characteristics of the breeding system of O. delavayi through the study of phenology, sex ratio, floral organ morphology, pollen number, stigma receptivity, artificial pollination, pollinators, and gene flow within and between populations, while also discussing the evolution and maintenance of androdioecy within the genus. The proportion of males was less than 0.5 and the out-crossing index (OCI) was 5. Morphological androdioecy was observed, with hermaphrodite flowers having fertile pistils, while male flowers had degenerated pistils. Males and hermaphrodites both had large amounts of small and fertile pollen grains, although the pollen number of males was ca. 1.21 × more than that of hermaphrodites, and pollen was generally smaller. Self-pollination was found to produce a much lower fruit set than outcrossing under natural conditions. Gene flow between males and hermaphrodites within a population was greater (1.007) than that between populations (0.753). All these results indicate that O. delavayi is functionally androdioecious, which may be an intermediate state in the evolutionary transition from hermaphroditism to dioecy.

Isohydric species are not necessarily more carbon limited than anisohydric species during drought.

Isohydry (i.e., strong regulation of leaf water potential, Ψl) is commonly associated with strict stomatal regulation of transpiration under drought, which in turn is believed to minimize hydraulic risk at the expense of reduced carbon assimilation. Hence, the iso/anisohydric classification has been widely used to assess drought resistance and mortality mechanisms across species, with isohydric species being hypothetically more prone to carbon starvation and anisohydric species more vulnerable to hydraulic failure. These hypotheses and their underlying assumptions, however, have rarely been tested under controlled, experimental conditions. Our objective is to assess the physiological mechanisms underlying drought resistance differences between two co-occurring Mediterranean forest species with contrasting drought responses: Phillyrea latifolia L. (anisohydric and more resistant to drought) and Quercus ilex L. (isohydric and less drought resistant). A total of 100 large saplings (50 per species) were subjected to repeated drought treatments for a period of 3 years, after which Q. ilex showed 18% mortality whereas no mortality was detected in P. latifolia. Relatively isohydric behavior was confirmed for Q. ilex, but higher vulnerability to cavitation in this species implied that estimated embolism levels were similar across species (12-52% in Q. ilex vs ~30% in P. latifolia). We also found similar seasonal patterns of stomatal conductance and assimilation between species. If anything, the anisohydric P. latifolia tended to show lower assimilation rates than Q. ilex under extreme drought. Similar growth rates and carbon reserves dynamics in both species also suggests that P. latifolia was as carbon-constrained as Q. ilex. Increasing carbon reserves under extreme drought stress in both species, concurrent with Q. ilex mortality, suggests that mortality in our study was not triggered by carbon starvation. Our results warn against making direct connections between Ψl regulation, stomatal behavior and the mechanisms of drought-induced mortality in plants.

The History of Attack and Success of Emerald Ash Borer (Coleoptera: Buprestidae) on White Fringetree in Southwestern Ohio.

Emerald ash borer, Agrilus planipennis Fairmaire, is an invasive insect that has caused widespread mortality of ash species in North America. The ability of emerald ash borer to utilize white fringetree as an alternate host was reported recently. We aimed to determine how long white fringetree has been under attack from emerald ash borer, the degree of attack, and the overall success of this beetle on this novel host. Stems from three of nine infested white fringetrees collected from the Dayton and Cincinnati, OH, areas in the winter of 2015 yielded four live adult emerald ash borers after being held in rearing containers, and numerous older exit holes were observed. Measurement and aging of feeding galleries on these stems indicated that emerald ash borer has been using this species since 2011, at least, with peak gallery densities reached in 2012 and 2013 on most of the harvested trees. On average, 32 galleries per square meter were found in these stems with about one-third of them being indicative of fourth-instar larvae. This supports the assertion that emerald ash borer has been using white fringetree as a host plant with moderate to good success for as long as ash species in these particular areas have been utilized.

Evidence for the long-term maintenance of a rare self-incompatibility system in Oleaceae.

A rare homomorphic diallelic self-incompatibility (DSI) system discovered in Phillyrea angustifolia (family Oleaceae, subtribe Oleinae) can promote the transition from hermaphroditism to androdioecy. If widespread and stable in Oleaceae, DSI may explain the exceptionally high rate of androdioecious species reported in this plant family. Here, we set out to determine whether DSI occurs in another Oleaceae lineage. We tested for DSI in subtribe Fraxininae, a lineage that diverged from subtribe Oleinae c. 40 million yr ago. We explored the compatibility relationships in Fraxinus ornus using 81 hermaphrodites and 25 males from one natural stand and two naturalized populations using intra- and interspecific stigma tests performed on F. ornus and P. angustifolia testers. We uncovered a DSI system with hermaphrodites belonging to one of two self-incompatibility (SI) groups and males compatible with both groups, making for a truly androdioecious reproductive system. The two human-founded populations contained only one of the two SI groups. Our results provide evidence for the evolutionary persistence of DSI. We discuss how its stability over time may have affected transitions to other sexual systems, such as dioecy.

Mesophyll conductance plays a central role in leaf functioning of Oleaceae species exposed to contrasting sunlight irradiance.

The ability to modify mesophyll conductance (gm ) in response to changes in irradiance may be a component of the acclimation of plants to shade-sun transitions, thus influencing species-specific distributions along light-gradients, and the ecological niches for the different species. To test this hypothesis we grew three woody species of the Oleaceae family, the evergreen Phillyrea latifolia (sun-requiring), the deciduous Fraxinus ornus (facultative sun-requiring) and the hemi-deciduous Ligustrum vulgare (shade tolerant) at 30 or 100% sunlight irradiance. We show that neither mesophyll conductance calculated with combined gas exchange and chlorophyll fluorescence techniques (gm) nor CO2 assimilation significantly varied in F. ornus because of sunlight irradiance. This corroborates previous suggestions that species with high plasticity for light requirements, do not need to undertake extensive reorganization of leaf conductances to CO2 diffusion to adapt to different light environments. On the other hand, gm steeply declined in L. vulgare and increased in P. latifolia exposed to full-sun conditions. In these two species, leaf anatomical traits are in part responsible for light-driven changes in gm , as revealed by the correlation between gm and mesophyll conductance estimated by anatomical parameters (gmA). Nonetheless, gm was greatly overestimated by gmA when leaf metabolism was impaired because of severe light stress. We show that gm is maximum at the light intensity at which plant species have evolved and we conclude that gm actually plays a key role in the sun and shade adaptation of Mediterranean species. The limits of gmA in predicting mesophyll conductance are also highlighted.

Contrasting impacts of continuous moderate drought and episodic severe droughts on the aboveground-biomass increment and litterfall of three coexisting Mediterranean woody species.

Climate change is predicted to increase the aridity in the Mediterranean Basin and severely affect forest productivity and composition. The responses of forests to different timescales of drought, however, are still poorly understood because extreme and persistent moderate droughts can produce nonlinear responses in plants. We conducted a rainfall-manipulation experiment in a Mediterranean forest dominated by Quercus ilex, Phillyrea latifolia, and Arbutus unedo in the Prades Mountains in southern Catalonia from 1999 to 2014. The experimental drought significantly decreased forest aboveground-biomass increment (ABI), tended to increase the litterfall, and decreased aboveground net primary production throughout the 15 years of the study. The responses to the experimental drought were highly species-specific. A. unedo suffered a significant reduction in ABI, Q. ilex experienced a decrease during the early experiment (1999-2003) and in the extreme droughts of 2005-2006 and 2011-2012, and P. latifolia was unaffected by the treatment. The drought treatment significantly increased branch litterfall, especially in the extremely dry year of 2011, and also increased overall leaf litterfall. The drought treatment reduced the fruit production of Q. ilex, which affected seedling recruitment. The ABIs of all species were highly correlated with SPEI in early spring, whereas the branch litterfalls were better correlated with summer SPEIs and the leaf and fruit litterfalls were better correlated with autumn SPEIs. These species-specific responses indicated that the dominant species (Q. ilex) could be partially replaced by the drought-resistant species (P. latifolia). However, the results of this long-term study also suggest that the effect of drought treatment has been dampened over time, probably due to a combination of demographic compensation, morphological and physiological acclimation, and epigenetic changes. However, the structure of community (e.g., species composition, dominance, and stand density) may be reordered when a certain drought threshold is reached.

Selfish male-determining element favors the transition from hermaphroditism to androdioecy.

According to the current, widely accepted paradigm, the evolutionary transition from hermaphroditism toward separate sexes occurs in two successive steps: an initial, intermediate step in which unisexual individuals, male or female, sterility mutants coexist with hermaphrodites and a final step that definitively establishes dioecy. Two nonexclusive processes can drive this transition: inbreeding avoidance and reallocation of resources from one sexual function to the other. Here, we report results of controlled crosses between males and hermaphrodites in Phillyrea angustifolia, an androdioecious species with two mutually intercompatible, but intraincompatible groups of hermaphrodites. We observed different segregation patterns that can be explained by: (1) epistatic interactions between two unlinked diallelic loci, determining sex and mating compatibility, and (2) a mutation with pleiotropic effects: female sterility, full compatibility of males with both hermaphrodite incompatibility groups, and complete male-biased sex-ratio distortion in one of the two groups. Modeling shows that these mechanisms can explain the high frequency of males in populations of P. angustifolia and can promote the maintenance of androdioecy without requiring inbreeding depression or resource reallocation. We thus argue that segregation distortion establishes the right conditions for the evolution of cryptic dioecy and potentially initiates the evolution toward separate sexes.

Incompatibility between plant-derived defensive chemistry and immune response of two sphingid herbivores.

Herbivorous insects use several different defenses against predators and parasites, and tradeoffs among defensive traits may occur if these traits are energetically demanding. Chemical defense and immune response potentially can interact, and both can be influenced by host plant chemistry. Two closely related caterpillars in the lepidopteran family Sphingidae are both attacked by the same specialist endoparasitoid species but have mostly non-overlapping host plant ranges that differ in secondary chemistry. Ceratomia catalpae is a specialist on Catalpa and also will feed on Chilopsis, which both produce iridoid glycosides. Ceratomia undulosa consumes members of the Oleaceae, which produce seco-iridoid glycosides. Immune response of the two species on a typical host plant species (Catalpa bignonioides for C. catalpa; Fraxinus americana for C. undulosa) was compared using a melanization assay, and did not differ. In a second experiment, the iridoid glycoside catalpol was added to the diets of both insects, and growth rate, mass, chemical defense, and immune response were evaluated. Increased dietary catalpol weakened the immune response of C. undulosa and altered the development rate of C. catalpae by prolonging the third instar and accelerating the fourth instar. Catalpol sequestration was negatively correlated with immune response of C. catalpae, while C. undulosa was unable to sequester catalpol. These results show that immune response can be negatively influenced by increasing concentrations of sequestered defensive compounds.

A one-locus model of androdioecy with two homomorphic self-incompatibility groups: expected vs. observed male frequencies.

Androdioecy, the occurrence of males and hermaphrodites in a single population, is a rare breeding system because the conditions for maintenance of males are restrictive. In the androdioecious shrub Phillyrea angustifolia, high male frequencies are observed in some populations. The species has a sporophytic self-incompatibility (SI) system with two self-incompatibility groups, which ensures that two groups of hermaphrodites can each mate only with the other group, whereas males can fertilize hermaphrodites of both groups. Here, we analyse a population genetic model to investigate the dynamics of such an androdioecious species, assuming that self-incompatibility and sex phenotypes are determined by a single locus. Our model confirms a previous prediction that a slight reproductive advantage of males relative to hermaphrodites allows the maintenance of males at high equilibrium frequencies. The model predicts different equilibria between hermaphrodites of the two SI groups and males, depending on the male advantage, the initial composition of the population and the population size, whose effect is studied through stochastic simulations. Although the model can generate high male frequencies, observed frequencies are considerably higher than the model predicts. We finally discuss how this model may help explain the large male frequency variation observed in other androdioecious species of Oleaceae: some species show only androdioecious populations, as P. angustifolia, whereas others show populations either completely hermaphrodite or androdioecious.

Xylem vulnerability to cavitation can be accurately characterised in species with long vessels using a centrifuge method.

Vulnerability to cavitation curves describe the decrease in xylem hydraulic conductivity as xylem pressure declines. Several techniques for constructing vulnerability curves use centrifugal force to induce negative xylem pressure in stem or root segments. Centrifuge vulnerability curves constructed for long-vesselled species have been hypothesised to overestimate xylem vulnerability to cavitation due to increased vulnerability of vessels cut open at stem ends that extend to the middle or entirely through segments. We tested two key predictions of this hypothesis: (i) centrifugation induces greater embolism than dehydration in long-vesselled species, and (ii) the proportion of open vessels changes centrifuge vulnerability curves. Centrifuge and dehydration vulnerability curves were compared for a long- and short-vesselled species. The effect of open vessels was tested in four species by comparing centrifuge vulnerability curves for stems of two lengths. Centrifuge and dehydration vulnerability curves agreed well for the long- and short-vesselled species. Centrifuge vulnerability curves constructed using two stem lengths were similar. Also, the distribution of embolism along the length of centrifuged stems matched the theoretical pressure profile induced by centrifugation. We conclude that vulnerability to cavitation can be accurately characterised with vulnerability curves constructed using a centrifuge technique, even in long-vesselled species.

Assessment of factors affecting micropropagation and ex vitro acclimatization of Nyctanthes arbor-tristis L.

Rapid differentiation of multiple shoots was observed in 94% of nodal explants of one year old Nyctanthes arbor-tristis L. plants. Shoot bud induction and multiplication took place on Murashige and Skoog (MS) medium supplemented with two cytokinins, i.e. Benzyladenine (BA) or Kinetin (Kn) either alone or in combination with different auxins, indole-3-butyric acid (IBA), indole-3-acetic acid (IAA) or α-naphthalene acetic acid (NAA). Between different media, pH levels and growth regulators tried, the optimum condition for maximum regenerative response was obtained on MS + Kn (2.5 µM) + N AA (0.5 µM) media at 5.8 pH, forming cultures with 23.26 ± 0.89 number of shoots and 6.36 ± 0.80 cm shoot length after 8 weeks of culture. Histological sections confirmed the formation of multiple buds from nodal explants. Rooting was achieved ex vitro by dipping the basal ends of microshoots in 200 µM IBA for 30 min followed by their transplantation in sterile soilrite. The plantlets with well-developed shoot and root system were successfully established in garden soil and grown outside in a greenhouse with a 80% survival rate.

A self-incompatibility system explains high male frequencies in an androdioecious plant.

Androdioecy is a sexual system in which males co-occur with hermaphrodites, which have both male and female function. Stable androdioecy is rare in nature, and theory suggests that it requires that males sire more than twice as many offspring as hermaphrodites. In several members of the olive family (Oleaceae), androdioecy occurs with higher frequencies of males than predicted by theory. In Phillyrea angustifolia L., we found that high male frequencies can be maintained in natural populations because hermaphrodites belong exclusively to one of two self-incompatibility groups, and thus, each can fertilize only half of all pollen recipients. In contrast, males can pollinate all hermaphrodites. Thus, in this species, the reproductive disadvantage that males face due to the loss of female function is offset by the fact that all males are fully compatible with all pollen recipients.

Seed trait changes in dispersers' guts and consequences for germination and seedling growth.

The effectiveness of a frugivore as a disperser of a plant is greatly determined by how fruits and seeds are handled in its mouth and its digestive tract. Although a number of studies have investigated the effect of avian ingestion on germination, we still know very little about the modifications to seeds during ingestion and the specific consequences on plant fitness. Here we investigate for the first time the different mechanisms by which germination patterns of seeds are modified following ingestion by frugivores. Specifically, we examine changes in seed mass, water content, permeability, seed coat thickness, texture, and resistance in two common Mediterranean fleshy-fruited plants, Phillyrea angustifolia and Myrtus communis, after ingestion by Eurasian Blackbirds, Turdus merula. We found a number of differences between the plant species: Phillyrea seeds lost mass, mainly due to water loss, and had thinner coats after gut passage, but Myrtus seeds did not. Seeds of both species showed increased permeability, while Myrtus seeds in particular became less resistant to breakage. No quantifiable changes in seed coat texture were detected in either species, although this trait was partly associated with differences in germination rate in Phillyrea. High intraspecific plant variation was found for most seed traits measured. Seed passage through birds' guts sped up germination in both species, especially in Myrtus. Increased permeability in seeds of both species following ingestion resulted in a higher germination rate. Moreover, seeds with thick coats (and in the case of Phillyrea, harder coats) germinated at a slower rate and produced seedlings that also grew more slowly, indicating a cost of coat thickness and/or hardness for seedling emergence. Results obtained here contribute to explaining the great heterogeneity in germination responses among and within plant species and the large variety of factors, both intrinsic and extrinsic to the plants, that influence such responses.

Below-ground interactions with arbuscular mycorrhizal shrubs decrease the performance of pinyon pine and the abundance of its ectomycorrhizas.

Few studies have examined how below-ground interactions among plants affect the abundance and community composition of symbiotic mycorrhizal fungi. Here, we combined observations during drought with a removal experiment to examine the effects of below-ground interactions with arbuscular mycorrhizal (AM) shrubs on the growth of pinyon pines (Pinus edulis), and the abundance and community composition of their ectomycorrhizal (EM) fungi. Shrub density was negatively correlated with pinyon above- and below-ground growth and explained 75% of the variation in EM colonization. Consistent with competitive release, pinyon fine-root biomass, shoot length and needle length increased with shrub removal. EM colonization also doubled following shrub removal. EM communities did not respond to shrub removal, perhaps because of their strikingly low diversity. These results suggest that below-ground competition with AM shrubs negatively impacted both pinyons and EM fungi. Similar competitive effects may be observed in other ecosystems given that drought frequency and severity are predicted to increase for many land interiors.

Quantitative study of fluorescence excitation and emission spectra of bean leaves.

A quantitative and comprehensive knowledge of leaf fluorescence is required for the interpretation of fluorescence signals at the canopy level and also for the modelling of leaf and canopy fluorescence. In this work we present full range fluorescence excitation and emission spectra of intact leaves, expressed in units of apparent spectral fluorescence yield, from both the adaxial and the abaxial sides of the leaves, and for both front-side and back-side geometries. Emission spectra were measured for incident radiations in the blue and the green spectral range. The red/far-red fluorescence ratio depended on the measurement geometry and on the excitation wavelength. Excitation spectra were measured for emissions at 687 and 760 nm. When the abaxial side was illuminated, the measured spectra always had a larger intensity compared to adaxial side that is explained by the higher scattering of the spongy tissues. At 760 nm, the spectra had the same shape for front-side and back-side geometry, indicating that scattering predominated. At 687 nm, the shape of the spectra was very different for front-side and back-side geometry due to re-absorption of red fluorescence within the leaf. The comparison of excitation spectra measured from the adaxial or the abaxial side revealed differences in carotenoid absorption.

Relative growth rate and biomass allocation in ten woody species with different leaf longevity using phylogenetic independent contrasts (PICs).

In this study, we compare the relative growth rate (RGR) and biomass allocation of 10 woody species (5 deciduous and 5 evergreen) from the Mediterranean region using phylogenetic independent contrasts (PICs) to test if these two functional groups differ in these traits. In general, the results were similar when using PICs or without taking into account phylogenetic relations. Deciduous species had a higher RGR than evergreen species, due to the higher net assimilation rate (NAR). Deciduous species had a higher specific leaf area (SLA) but a lower leaf mass ratio (LMR), resulting in a similar LAR for deciduous and evergreen species (LAR = SLA x LMR). In some cases, the use of PICs revealed patterns that would not have appeared if phylogeny had been overlooked. For example, there was no significant correlation between RGR and final dry mass (after 4 months of growth) but PICs revealed that there was a positive relation between these two variables in all deciduous-evergreen pairs. In general, RGR decreased with time and this temporal variation was due primarily to NAR variations (r = 0.79, p < 0.01), and also to variations in LAR (r = 0.69, p < 0.05). Considering the phylogeny, the only variable constantly different for all deciduous-evergreen pairs was SLA. This result, and the fact that SLA was the best correlated variable with RGR (r = 0.81, p < 0.01), reinforce the value of SLA as a variable closely associated to growth and to the functional groups (deciduous vs. evergreen).

Tissue-water relations of two co-occurring evergreen Mediterranean species in response to seasonal and experimental drought conditions.

Tissue-water relations were used to characterize the responses of two Mediterranean co-occurring woody species (Quercus ilex L. and Phillyrea latifolia L.) to seasonal and experimental drought conditions. Soil water availability was reduced approximately 15% by partially excluding rain throughfall and lateral flow (water runoff). Seasonal and experimental drought elicited physiological and morphological adaptations other than osmotic adjustment: both species showed large increases in cell-wall elasticity and decreased saturated-to-dry-mass ratio. Increased elasticity (lower elastic modulus) resulted in concurrent decreases in relative water content at turgor loss. In addition, P. latifolia showed significant increases in apoplastic water fraction. Decreased saturated-to-dry-mass ratio and increased apoplastic water fraction were accompanied by an increased range of turgor maintenance, which indicates that leaf sclerophyllous traits might be advantageous in drier scenarios. In contrast, the degree of sclerophylly (as assessed by the leaf mass-to-area ratio) was not related to tissue elasticity. An approximately 15% reduction in soil water availability resulted in significant reductions in diameter growth when compared to control plants in both species. Moreover, although P. latifolia underwent larger changes in tissue water-related traits than Q. ilex in response to decreasing water availability, growth was more sensitive to water stress in P. latifolia than in Q. ilex. Differences in diameter growth between species might be partially linked to the effects of cell-wall elasticity and turgor pressure on growth, since Q. ilex showed higher tissue elasticity and higher intrinsic tolerance to water deficit (as indicated by lower relative water content at turgor loss) than P. latifolia.

Storage behavior of Chionanthus retusus seed and asynchronous development of the radicle and shoot apex during germination in relation to germination inhibitors, including abscisic acid and four phenolic glucosides.

Studies on seed storage of Chionanthus retusus Lindl. & Paxt. revealed an orthodox behavior, one which showed both desiccation and freezing tolerance. An epicotyl after-ripening dormancy was expressed in C. retusus seeds by slow growth of the shoot apex relative to more rapid growth of the radicle when seeds were germinated at 30/20 degrees C. Although these seeds exhibit radicle protrusion, they must be after-ripened for another 8-10 weeks at 30/20 degrees C in order to obtain normal shoot growth. Removal of the endosperm, however, quickly stimulated cotyledon and shoot emergence without the additional after-ripening. Water-soluble glucoside phenolics, GL-3, Nuzhenide, ligustroside and oleoside dimethyl ester are present at relatively high levels in endosperm of freshly harvested seeds. These glucoside phenolics are excreted from the endosperm during subsequent after-ripening. Embryo and endosperm tissue from seed germinating at 30/20 degrees C (germination being defined by protrusion of the radicle) had a 10 times lower abscisic acid (ABA) content than similar tissues from freshly harvested mature seed. However, no shoot growth occurred even with the 10-fold reduction in ABA and a concomitant increase in endogenous gibberellins A1, A4 and A20. Thus, epicotyl dormancy during the first 8 weeks of after-ripening at 30/20 degrees C may be controlled by factors other than high ABA, i.e., the slow development of the shoot apex following radicle protrusion may be controlled more by high levels of glucoside phenolics than by diminished ABA and elevated GA levels.

Sap flow of three co-occurring Mediterranean woody species under varying atmospheric and soil water conditions.

We studied the seasonal patterns of water use in three woody species co-occurring in a holm oak forest in northeastern Spain. The three species studied, Quercus ilex L., Phillyrea latifolia L. and Arbutus unedo L., constitute more than 99% of the total basal area of the forest. The study period included the dry seasons of 1999 and 2000. Water use was estimated with Granier-type sap flux sensors. Standard meteorological variables, soil water content and leaf water potentials were also monitored. All monitored individuals reduced leaf-related sap flow (Q(l)) during the summer, concurrent with an increase in soil moisture deficit (SMD). Despite similar maximum Q(l) between species, the decline in Q(l) with increasing SMD was species-dependent. The average reduction in Q(l) between early summer and the peak of the drought was 74% for A. unedo (n = 3), 58% for P. latifolia (n = 3) and 87% for Q. ilex (n = 1). The relationship between canopy stomatal conductance (G(s)) and vapor pressure deficit (D) changed during the course of the drought, with progressively lower G(s) for any given D. Summertime reductions of Q(l) and G(s) were associated with between-species differences in vulnerability to xylem embolism, and with the corresponding degree of native embolism (lowest in P. latifolia and highest in Q. ilex). Our results, combined with previous studies in the same area, outlined differences among the species studied in manner of responding to water shortage, with P. latifolia able to maintain water transport at much lower water potentials than the other two species. In an accompanying experiment, A. unedo responded to an experimental reduction in water availability by reducing Q(l) during the summer. This species also modified its water use between years according to the different seasonal patterns of precipitation. These results are discussed in relation to the possible impacts that climate change will have on Q. ilex-dominated forests.