ALBERT

Description: The Mediterranean region is a hot spot of climate change vulnerable to increased droughts and heat waves. Scaling carbon fluxes from leaf to landscape levels is particularly challenging under drought conditions. We aimed to improve the mechanistic understanding of the seasonal acclimation of photosynthesis and morphology in sunlit and shaded leaves of four Mediterranean trees ( Quercus ilex L., Pinus halepensis Mill., Arbutus unedo L. and Quercus pubescens Willd.) under natural conditions. V c,max and J max were not constant, and mesophyll conductance was not infinite, as assumed in most terrestrial biosphere models, but varied significantly between seasons, tree species and leaf position. Favourable conditions in winter led to photosynthetic recovery and growth in the evergreens. Under moderate drought, adjustments in the photo/biochemistry and stomatal/mesophyllic diffusion behaviour effectively protected the photosynthetic machineries. Severe drought, however, induced early leaf senescence mostly in A. unedo and Q. pubescens , and significantly increased leaf mass per area in Q. ilex and P. halepensis . Shaded leaves had lower photosynthetic potentials but cushioned negative effects during stress periods. Species-specificity, seasonal variations and leaf position are key factors to explain vegetation responses to abiotic stress and hold great potential to reduce uncertainties in terrestrial biosphere models especially under drought conditions.

Description: Plants experiencing drought stress are frequently more susceptible to pathogens, likely via alterations in physiology that create favorable conditions for pathogens. Common plant responses to drought include the production of reactive oxygen species (ROS) and the accumulation of free amino acids (AAs), particularly proline. These same phenomena also frequently occur during pathogenic attack. Therefore, drought-induced perturbations in AA and ROS metabolism could potentially contribute to the observed enhanced susceptibility. Furthermore, nitrogen (N) availability can influence AA accumulation and affect plant resistance, but its contributions to drought-induced susceptibility are largely unexplored. Here we show that drought induces accumulation of hydrogen peroxide (H 2 O 2 ) in Austrian pine ( Pinus nigra Arnold) shoots, but that shoot infection by the blight and canker pathogen Diplodia sapinea (Fr.) Fuckel leads to large reductions in H 2 O 2 levels in droughted plants. In in vitro assays, H 2 O 2 was toxic to D. sapinea , and the fungus responded to this oxidative stress by increasing catalase and peroxidase activities, resulting in substantial H 2 O 2 degradation. Proline increased in response to drought and infection when examined independently, but unlike all other AAs, proline further increased in infected shoots of droughted trees. In the same tissues, the proline precursor, glutamate, decreased significantly. Proline was found to protect D. sapinea from H 2 O 2 damage, while also serving as a preferred N source in vitro. Fertilization increased constitutive and drought-induced levels of some AAs, but did not affect plant resistance. A new model integrating interactions of proline and H 2 O 2 metabolism with drought and fungal infection of plants is proposed.

Description: Small differences in the sensitivity of stomatal conductance to light intensity on leaf surfaces may lead to large differences in total canopy transpiration ( E C ) with increasing canopy leaf area ( L ). Typically, the increase of L would more than compensate for the decrease of transpiration per unit of leaf area ( E L ), resulting in concurrent increase of E C . However, highly shade-intolerant species, such as Larix principis-rupprechtii Mayr., may be so sensitive to increased shading that such compensation is not complete. We hypothesized that in such a stand, windfall-induced spatial variation at a decameter scale would result in greatly reduced E L in patches of high L leading to lower E C than low competition patches of sparse canopy. We further hypothesized that quicker extraction of soil moisture in patches of lower competition will result in earlier onset of drought symptoms in these patches. Thus, patches of low L will transition from light to soil moisture as the factor dominating E L . This process should progressively homogenize E C in the stand even as the variation of soil moisture is increasing. We tested the hypotheses utilizing sap flux of nine trees, and associated environmental and stand variables. The results were consistent with only some of the expectations. Under non-limiting soil moisture, E L was very sensitive to the spatial variation of L , decreasing sharply with increasing L and associated decrease of mean light intensity on leaf surfaces. Thus, under the conditions of ample soil moisture maximum E C decreased with increasing patch-scale L . Annual E C and biomass production also decreased with L , albeit more weakly. Furthermore, variation of E C among patches decreased as average stand soil moisture declined between rain events. However, contrary to expectation, high L plots which transpired less showed a greater E L sensitivity to decreasing stand-scale soil moisture, suggesting a different mechanism than simple control by decreasing soil moisture. We offer potential explanations to the observed phenomenon. Our results demonstrate that spatial variation of L at decameter scale, even within relatively homogeneous, single-species, even-aged stands, can produce large variation of transpiration, soil moisture and biomass production and should be considered in 1-D soil–plant–atmosphere models.

Description: The main goal of this study was to develop a method for the extraction and indirect estimation of the quantity of calcium oxalate (CaOx) in the foliage of trees. Foliar tissue was collected from a single tree of each species (five conifers and five hardwoods) for comparison of extractions in different solvents using 10 replicates per species from the same pool of tissue. For each species, calcium (Ca) and oxalate were extracted sequentially in double deionized water and 2N acetic acid, and finally, five replicate samples were extracted in 5% (0.83N) perchloric acid (PCA) and the other five in 2N hydrochloric acid (HCl); three cycles of freezing and thawing were used for each solvent. Total ions were extracted by microwave digestion. Calcium was quantified with an inductively coupled plasma emission spectrophotometer method and oxalate was eluted and quantified using a high performance liquid chromatography method. This experiment was repeated again with two conifer and two hardwood species using four trees per species, and two analytical replicates for each tree. We report here that, regardless of age of individual trees within a species, time of collection or species type, the third extraction in PCA or HCl resulted in near equimolar quantities of Ca and oxalate ( r 2  ≥ 0.99). This method provides an easy estimate of the quantity of CaOx crystals using a small sample of foliar tissue. An additional benefit of PCA is that it precipitates the nucleic acids and proteins, allowing the quantification of several free/soluble metabolites such as amino acids, polyamines, organic acids and inorganic elements all from a single sample extract.

Description: Although copepods have been considered tolerant against the direct influence of the ocean acidification (OA) projected for the end of the century, some recent studies have challenged this view. Here, we have examined the direct impact of short-term exposure to a pCO 2 / pH level relevant for the year 2100 ( pH NBS , control: 8.18, low pH : 7.78), on the physiological performance of two representative marine copepods: the calanoid Acartia grani and the cyclopoid Oithona davisae . Adults of both species, from laboratory cultures, were preconditioned for four consecutive days in algal suspensions ( Akashiwo sanguinea ) prepared with filtered sea water pre-adjusted to the targeted pH values via CO 2 bubbling. We measured the feeding and respiratory activity and reproductive output of those pre-conditioned females. The largely unaffected fatty acid composition of the prey offered between OA treatments and controls supports the absence in the study of indirect OA effects (i.e. changes of food nutritional quality). Our results show no direct effect of acidification on the vital rates examined in either copepod species. Our findings are compared with results from previous short- and long-term manipulative experiments on other copepod species.

Description: We analyzed the genetic structure of the radiolarian morphospecies Larcopyle buetschlii from the surface to deep waters (up to 2000 m) in the Japan Sea using the internal transcribed spacer (ITS) regions of ribosomal RNA genes. Each individual had several ITS variants, but these polymorphisms show no vertical phylogeographic structure, suggesting a single biological species. Its rapid clonal reproduction suggested by high ITS variation likely plays a pivotal role in maintaining its wide vertical distribution.

Description: Phronima sedentaria is a hyperiid amphipod that diel migrates into a pronounced oxygen minimum zone (OMZ) in the Eastern Tropical North Pacific. In this study, oxygen consumption and lactate production were measured in P. sedentaria to estimate the aerobic and anaerobic contributions to total metabolism under conditions that mimic its day- (1% oxygen, 10°C) and night-time (20% oxygen, 20°C) habitat. When exposed to hypoxia and low temperature, the total metabolism of P. sedentaria was depressed by 78% compared with normoxic conditions. The metabolic enzymes citrate synthase (CS) and lactate dehydrogenase (LDH) were also measured as indicators of aerobic and anaerobic metabolism, and compared with specimens collected from the California Current and the North Atlantic to assess potential adaptations to low oxygen. LDH activity was not significantly different between regions. Significant differences in CS activity may be due to variation in food availability. Climate change is predicted to increase surface temperatures and cause the expansion of OMZs. This will result in vertical compression of the night-time range for P. sedentaria and is likely to have the same impact on other diel migrators. Habitat compression will reduce zooplankton contribution to carbon cycling and alter oceanic ecology, including predator–prey interactions.

Description: Many phytoplankton exploit phosphorus (P) from organic sources when dissolved inorganic P (DIP) is depleted. This process is, however, rarely considered in ecological and biogeochemical models. We present a mechanistic model describing explicitly the ability of phytoplankton to use dissolved organic P (DOP) when DIP is limiting, by synthesizing alkaline phosphatase (AP) that releases DIP from DOP. This model, applicable to any phytoplankton species expressing AP, is here specifically developed for the colony-forming Phaeocystis globosa. It describes the main processes related to P metabolism, including DIP transport, intracellular accumulation and assimilation. Model behaviour is explored in DIP-limiting batch-type conditions for different DOP ranging between 0 and 1.5 mmol P m –3 . Simulations show that the DOP-derived DIP increases the maximum biomass reached and extends the period of net growth. The magnitude of the enhanced biomass production is controlled by the DOP initially present as well as the released DOP, the latter being recycled by lysis of P. globosa cells. We also present a simplified model version derived from the mechanistic model, which involves fewer state variables and parameters. The latter is directly usable in both variable (quota-type) and fixed stoichiometry descriptions of phytoplankton growth.

Description: Zooplankton diel vertical migration (DVM) is often explained as a balance between predator avoidance and resource acquisition. However, recent studies suggest that ultraviolet radiation (UV) may also be important in driving zooplankton DVM in some systems. Williamson et al. ( Williamson et al ., 2011 ) proposed the "transparency-regulator hypothesis," which integrates UV into our current understanding of the drivers of DVM and predicts that the relative roles of UV and visual predation pressure will vary systematically across a gradient of lake transparency. To assess this hypothesis, we conducted in situ mesocosm experiments in five different lakes: two lakes without fish and three lakes with fish that spanned a range of UV and visible light transparency. We used an open-bottomed mesocosm design that allowed for the direct manipulation of UV that did not constrain visual predators or the amplitude or timing of natural migrations. Consistent with the transparency-regulator hypothesis, we found that UV is an important driver of Daphnia DVM in highly UV transparent lakes with and without fish but not in low transparency systems. Our results also suggest that UV and visual predation pressure may interact in systems of intermediate transparency.

Description: Competition for resources can lead to species exclusion. However, this exclusion may be avoided if species show differential adaptation to physical environment. Empirical studies on competition are difficult when species are phylogenetically close and have complex life cycles. This is the case of B. plicatilis and B. manjavacas , two cryptic rotifer species differing in their salinity niches and in life-history traits related to sex and diapause. These differences have been suggested to promote the stable co-occurrence observed in natural populations of these species. However, in a previous empirical study, the outcome of competition between both species was always exclusion. Here, we theoretically explored the effect of complex life-history traits and salinity fluctuations on the long-term competitive outcome of B. plicatilis and B. manjavacas . We developed a model and simulated ecological scenarios combining different growing period lengths, levels of crossed induction of sex between species and salinity regimes. Results show that a fluctuating salinity regime, an intermediate length of growing season and a low level of crossed induction of sex are essential conditions to take into account to explain coexistence.

Description: Colonization of new habitats through dispersal of phytoplankton cysts might be limited, if resident populations outcompete invaders during germination. We reciprocally transferred Gonyostomum semen (Raphidophyceae) cysts from three lakes into native and foreign waters originating from the respective habitats. Germination rate and germling growth were impacted by water origin, but there was no preference for native water. Gonyostomum semen 's ability to germinate in different conditions might explain its expansion in northern Europe.

Description: Measuring zooplankton biomass and physiological rates is of paramount importance in biological oceanography in order to assess the role of this community in, e.g. carbon fluxes. Classical methods (incubations) are very time-consuming and cannot match the frequency of physical and chemical measurements. Attempting to solve this, a variety of methods (e.g. egg production, RNA/DNA ratio or enzyme activities) have been developed over the last decades. These methods also show uncertainties and hitherto only incubation methods have been widely accepted. Predictive equations relating physiological processes and body weight (bw) and temperature are a rough alternative, normally used to ascertain the role of these organisms in the oceanic ecosystem. However, using imaging systems and empirical relationships to determine bw allows the application of physiological models to each individual, obtaining reliable estimates for taxonomic groups and size classes. In this study, we developed predictive equations suitable for growth and respiration estimations in subtropical regions. In addition, biomass and physiological rates assessed from empirical equations in combination with an image-based system (ZooImage) were compared with standard and enzymatic methods, respectively. We observed a consistent agreement between methodologies, the former resulting in an inexpensive and faster procedure for the appraisal of biomass and community carbon fluxes at large spatial and temporal scales.

Description: Carbon-specific prey clearance and ingestion rates of 1.5-mm tentaculate larvae of the ctenophore Mnemiopsis leidyi increased linearly between 6 and 25°C but declined between 25 and 30°C. Both absolute (length) and carbon-specific growth rate increased linearly with increasing temperature. The latter was 0.87 d –1 at 25°C. Extremely low or negative growth rates observed at 6 and 30°C help define the thermal limits to population growth of this successful biological invader.

Description: We have supplemented available, concurrent measurements of fresh weight ( W , g) and body carbon (C, g) (46 individuals, 14 species) and nitrogen (N, g) (11 individuals, 9 species) of marine gelatinous animals with data obtained during the global ocean MALASPINA 2010 Expedition (totalling 267 individuals and 33 species for the W versus C data; totalling 232 individuals and 31 species for the N versus C data). We then used those data to test the allometric properties of the W versus C and N versus C relationships. Overall, gelatinous organisms contain 1.13 ± 1.57% of C (by weight, mean ± SD) in their bodies and show a C:N of 4.56 ± 2.46, respectively, although estimations can be improved by using separate conversion coefficients for the carnivores and the filter feeders. Reduced major axis regression indicates that W increases isometrically with C in the carnivores (cnidarians and ctenophores), implying that their water content can be described by a single conversion coefficient of 173.78 gW(g C) –1 , or a C content of 1.17 ± 1.90% by weight, although there is much variability due to the existence of carbon-dense species. In contrast, W increases more rapidly than C in the filter feeders (salps and doliolids), according to a power relationship W = 446.68C 1.54 . This exponent is not significantly different from 1.2, which is consistent with the idea that the watery bodies of gelatinous animals represent an evolutionary response towards increasing food capture surfaces, i.e. a bottom-up rather than a top-down mechanism. Thus, the available evidence negates a bottom-up mechanism in the carnivores, but supports it in the filter feeders. Last, N increases isometrically with C in both carnivores and filter feeders with C:N ratios of 3.89 ± 1.34 and 4.38 ± 1.21, respectively. These values are similar to those of compact, non-gelatinous organisms and reflect a predominantly herbivorous diet in the filter feeders, which is confirmed by a difference of one trophic level between filter feeders and carnivores, according to stable N isotope enrichment data.

Description: I explored mortality estimation for stage-structured populations, building on previous work that applied vertical life-table methods to populations of copepods. A new Bayesian approach for estimating mortality rates accounts for uncertainties in stage duration and number counted by stage, which have not been fully incorporated into previous analyses. This method assumes that mortality is similar among similar life stages. Results using simulated data show that realistic values of the standard deviation of stage duration and number of individuals counted result in reliable mortality estimates, though with wide confidence intervals. This uncertainty obscures variation in estimated mortality between successive stages and can also obscure bias due to violation of underlying assumptions such as that of a stable stage distribution. More importantly, the uncertainty calls into question many previous mortality estimates across pairs of life stages that do not account for these sources of uncertainty. The method was applied to an introduced population of the brackish-water cyclopoid copepod Limnoithona tetraspina in the San Francisco Estuary. Despite the uncertainties, results were interpretable: mortality was highest in nauplii and lowest in adults, probably because of high vulnerability of nauplii to invertebrate predators and low vulnerability of adults to fish predation.

Description: The strong La Niña of 2010–2011 provided an opportunity to investigate the ecological impacts of El Niño-Southern Oscillation on coastal plankton communities using the nine national reference stations around Australia. Based on remote sensing and across the entire Australian region 2011 (La Niña) was only modestly different from 2010 (El Niño) with the average temperature declining 0.2%, surface chlorophyll a up 3% and modelled primary production down 14%. Other changes included a poleward shift in Prochlorococcus and Synechococcus . Along the east coast, there was a reduction in salinity, increase in nutrients, Chlorophytes and Prasinophytes (taxa with chlorophyll b , neoxanthin and prasinoxanthin). The southwest region had a rise in the proportion of 19-hexoyloxyfucoxanthin; possibly coccolithophorids in eddies of the Leeuwin Current and along the sub-tropical front. Pennate diatoms increased, Ceratium spp. decreased and Scrippsiella spp. increased in 2011. Zooplankton biomass declined significantly in 2011. There was a reduction in the abundance of Calocalanus pavo and Temora turbinata and increases in Clausocalanus farrani , Oncaea scottodicarloi and Macrosetella gracilis in 2011. The changes in the plankton community during the strong La Niña of 2011 suggest that this climatic oscillation exacerbates the tropicalization of Australia.

Description: Jellyfish are effective predators on mesozooplankton and release large amounts of dissolved organic matter. Nevertheless, jellyfish initiated trophic cascades and bottom-up influences impacting lower trophic levels have received limited attention. We conducted a mesocosm experiment to quantify simultaneous top-down and bottom-up effects of a common jellyfish, Cyanea capillata , in a natural plankton community during autumn. Treatments were 0, 2 or 5 jellyfish per 2.5 m 3 mesocosm, four replicates each, with initial additions of inorganic nutrients. Primary and bacterial production, species abundance and composition of several trophic levels and nutrient and carbon dynamics were followed during the 8-day experiment. Multivariate statistics and generalized additive mixed modelling were applied to test whether jellyfish carbon concentration (0–1.26 mg jellyC L –1 ) in the mesocosms affected the variables monitored. Unexpected negligible predatory impact of jellyfish on mesozooplankton was observed, potentially related to jellyfish senescence. Community compositions of bacteria, phytoplankton and mesozooplankton changed with time, but did not differ between treatments. However, nutrient regeneration by jellyfish was evident, and jellyfish had a positive impact on total and specific bacterial production, total primary production and the 〉10 µm chlorophyll a fraction. Bottom-up influences from abundant jellyfish could thus stimulate productivity in nutrient depleted autumnal surface waters.

Description: The feeding ecology of Blackfordia virginica was evaluated concurrently with their ecophysiological condition in a temperate estuary. The diet of B. virginica is composed not only of metazooplankton, as commonly observed for other jellyfish species, but also of phytoplankton, ciliates and detritus. This feeding behavior might explain their good nutritional condition and sustainable growth during bloom peaks, when zooplankton abundance has already decreased significantly.

Description: The pelagic dynamics of the cosmopolitan scyphozoan Aurelia sp . was investigated in three French Mediterranean lagoons, Thau, Berre and Bages-Sigean, which harbour resident populations. The annual cycles showed a common univoltine pattern in all lagoons where the presence of pelagic stages in the water column lasted ~8 months. Field observations showed a release of ephyrae in winter time followed by pronounced growth between April and July, when individuals reached the largest sizes, before disappearing from the water column. Maximum abundance of ephyrae and medusae were registered in Thau. Medusae abundance attained a maximum of 331 ind 100 m –3 in Thau, 18 ind 100 m –3 in Berre and 7 ind 100 m –3 in Bages-Sigean lagoons. Temperature and zooplankton abundance appeared as leading factors of growth, where Bages-Sigean showed the population with higher growth rates (2.66 mm day –1 ) and maximum size (32 cm), followed by Thau (0.57–2.56 mm day –1 ; 22.4 cm) and Berre (1.57–2.22 mm day –1 ; 17 cm). The quantification of environmental windows used by the species showed wider ranges than previously reported in the Mediterranean Sea, which suggests a wide ecological plasticity of Aurelia spp. populations in north-western Mediterranean lagoons.

Description: Two lobate ctenophores, Bolinopsis infundibulum and Mnemiopsis leidyi , occur in the North Sea. Stomach contents of field-collected B. infundibulum were recorded and clearance rates for cladocerans and copepods calculated. In starvation experiments, daily body carbon losses of 2.2 and 1.2% and total carbon content losses of 76 and 63% were observed for B. infundibulum (after 68 days) and M. leidyi (after 67 days), respectively.

Description: The ctenophore Mnemiopsis leidyi is characterized by high growth rates and a large reproductive capacity. However, reproductive dynamics are not yet well understood. Here, we present laboratory data on food-dependent egg production in M. leidyi and egg hatching time and success. Further, we report on the reproduction of laboratory-reared and field-caught animals during starvation. Our results show that the half-saturation zooplankton prey concentration for egg production is reached at food levels of 12–23 µgC L –1 , which is below the average summer food concentration encountered in invaded areas of northern Europe. Furthermore, starved animals continue to produce eggs for up to 12 days after cessation of feeding with high overall hatching success of 65–90%. These life history traits allow M. leidyi to thrive and reproduce in environments with varying food conditions and give it a competitive advantage under unfavourable conditions. This may explain why recurrent population blooms are observed and sustained in localized areas in invaded northern Europe, where water exchange is limited and zooplankton food resources are quickly depleted by M. leidyi . We suggest that these reproductive life history traits are key to its invasion success.

Description: Surface-dwelling colonies of Velella velella occur throughout tropical to cold-temperate oceans of the world and sometimes are stranded in masses along hundreds of kilometers of beaches. Large-scale blooms in the Western Mediterranean Sea in 2013 and 2014 allowed the study of diet, prey digestion times and predation rates. Gastrozooid content analyses showed that 59% of the 769 identified prey were euphausiid larvae (calytopsis and furcilia) captured at night. Copepods (41%), fish eggs (2.2%) and larvae (0.5%) were captured both at day and night. Digestion times at ambient temperature (~17°C) of calytopsis, furcilia and copepods were estimated to be 〉6.5, 4.4 and 3.9 h, respectively. Estimated prey consumption was substantially lower in 2014 than in 2013 (41 vs. 75 prey day –1 colony –1 ). Velella velella and other gelatinous species bloomed in the Mediterranean Sea and the northeastern Atlantic and Pacific oceans in 2013 and 2014. Because of the wide distribution of V. velella colonies, their mass occurrences, potential importance as predators and competitors of fish, additional production from symbiotic zooxanthellae and stranding on beaches, they could be important in open-ocean carbon cycling and in transport of pelagic production to landmasses.

Description: The white-rot fungus Heterobasidion parviporum Niemelä & Korhonen establishes a necrotrophic interaction with Norway spruce ( Picea abies (L.) H.Karst.) causing root and butt rot and growth losses in living trees. The interaction occurs first with the bark and the outer sapwood, as the pathogen enters the tree via wounds or root-to-root contacts. Later, when the fungus reaches the heartwood, it spreads therein creating a decay column, and the interaction mainly occurs in the inner sapwood where the tree creates a reaction zone. While bark and outer sapwood interactions are well studied, little is known about the nature of the transcriptional responses leading to the creation of a reaction zone. In this study, we sampled bark and sapwood both proximal and distal to the reaction zone in artificially inoculated and naturally infected trees. We quantified gene expression levels of candidate genes in secondary metabolite, hormone biosynthesis and signalling pathways using quantitative polymerase chain reaction. An up-regulation of mainly the phenylpropanoid pathway and jasmonic acid biosynthesis was found at the inoculation site, when inoculations were compared with wounding. We found that transcriptional responses in inner sapwood were similar to those reported upon infection through the bark. Our data suggest that the defence mechanism is induced due to direct fungal contact irrespective of the tissue type. Understanding the nature of these interactions is important when considering tree breeding-based resistance strategies to reduce the spread of the pathogen between and within trees.

Description: Global warming and associated decreases in summer rainfall may threaten tree vitality and forest productivity in many regions of the temperate zone in the future. One option for forestry to reduce the risk of failure is to plant genotypes which combine high productivity with drought tolerance. Growth experiments with provenances from different climates indicate that drought exposure can trigger adaptive drought responses in temperate trees, but it is not well known whether and to what extent regional precipitation reduction can increase the drought resistance of a species. We conducted a common garden growth experiment with five European beech ( Fagus sylvatica L.) populations from a limited region with pronounced precipitation heterogeneity (816–544 mm year –1 ), where phylogenetically related provenances grew under small to large water deficits. We grew saplings of the five provenances at four soil moisture levels (dry to moist) and measured ~30 morphological (leaf and root properties, root : shoot ratio), physiological (leaf water status parameters, leaf conductance) and growth-related traits (above- and belowground productivity) with the aim to examine provenance differences in the drought response of morphological and physiological traits and to relate the responsiveness to precipitation at origin. Physiological traits were more strongly influenced by provenance (one-third of the studied traits), while structural traits were primarily affected by water availability in the experiment (two-thirds of the traits). The modulus of leaf tissue elasticity reached much higher values late in summer in plants from moist origins resulting in more rapid turgor loss and a higher risk of hydraulic failure upon drought. While experimental water shortage affected the majority of morphological and productivity-related traits in the five provenances, most parameters related to leaf water status were insensitive to water shortage. Thus, plant morphology, and root growth in particular, did respond to reduced water availability with higher phenotypic plasticity than did physiology. We conclude that beech provenances exposed to different precipitation regimes have developed some genotypic differences with respect to leaf water status regulation, but these adaptations are associated with only minor adaptation in plant morphology and they do not affect the growth rate of the saplings.

Description: Many studies have demonstrated linkages between the occurrence of fog and ecophysiological functioning in cloud forests, but few have investigated hydraulic functioning as a determining factor that explains sharp changes in vegetation. The objective of this study was to compare the plant water status during cloud-immersed and non-immersed conditions and hydraulic vulnerability in branches and roots of species across a temperate, mountain fog ecotone. Because cloud forests are often dark, cool and very moist, we expected cloud forest species to have less drought-tolerant characteristics (i.e., lower P e and P 50 —the pressures required to induce a 12 and 50% loss in hydraulic conductivity, respectively) relative to non-cloud forest species in adjacent (lower elevation) forests. Additionally, due to the ability of cloud forest species to absorb cloud-fog water, we predicted greater improvements in hydraulic functioning during fog in cloud forest species relative to non-cloud forest species. Across the cloud forest ecotone, most species measured were very resistant to losses in conductivity with branch P 50 values from –4.5 to –6.0 MPa, hydraulic safety margins ( min – P 50 ) 〉1.5 MPa and low calculated hydraulic conductivity losses. Roots had greater vulnerabilities, with P 50 values ranging from –1.4 to –2.5 MPa, leading to greater predicted losses in conductivity (~20%). Calculated values suggested strong losses of midday leaf hydraulic conductance in three of the four species, supporting the hydraulic segmentation hypothesis. In both cloud forest and hardwood species, s were greater on foggy days than sunny days, demonstrating the importance of fog periods to plant water balance across fog regimes. Thus, frequent fog did not result in systemic changes in hydraulic functioning or vulnerability to embolism across our temperate cloud forest ecotone. Finally, roots functioned with lower hydraulic conductivity than branches, suggesting that they may serve as more sensitive indicators of hydraulic functioning in these mesic, foggy ecosystems.

Description: Climate warming is having an impact on distribution, acclimation and defence capability of plants. We compared the emission rate and composition of volatile organic compounds (VOCs) from silver birch ( Betula pendula (Roth)) provenances along a latitudinal gradient in a common garden experiment over the years 2012 and 2013. Micropropagated silver birch saplings from three provenances were acquired along a gradient of 7° latitude and planted at central (Joensuu 62°N) and northern (Kolari 67°N) sites. We collected VOCs emitted by shoots and assessed levels of herbivore damage of three genotypes of each provenance on three occasions at the central site and four occasions at the northern site. In 2012, trees of all provenances growing at the central site had higher total VOC emission rates than the same provenances growing at the northern site; in 2013 the reverse was true, thus indicating a variable effect of latitude. Trees of the southern provenance had lower VOC emission rates than trees of the central and northern provenances during both sampling years. However, northward or southward translocation itself had no significant effect on the total VOC emission rates, and no clear effect on insect herbivore damage. When VOC blend composition was studied, trees of all provenances usually emitted more green leaf volatiles at the northern site and more sesquiterpenes at the central site. The monoterpene composition of emissions from trees of the central provenance was distinct from that of the other provenances. In summary, provenance translocation did not have a clear effect in the short-term on VOC emissions and herbivory was not usually intense at the lower latitude. Our data did not support the hypothesis that trees growing at lower latitudes would experience more intense herbivory, and therefore allocate resources to chemical defence in the form of inducible VOC emissions.

Description: Virus-induced gene silencing (VIGS) has been shown to be an effective tool for investigating gene functions in herbaceous plant species, but has rarely been tested in trees. The establishment of a fast and reliable transformation system is especially important for woody plants, many of which are recalcitrant to transformation. In this study, we established a tobacco rattle virus (TRV)-based VIGS system for two Populus species, Populus euphratica and P.   x   canescens . Here, TRV constructs carrying a 266 bp or a 558 bp fragment of the phytoene desaturase (PDS) gene were Agrobacterium -infiltrated into leaves of the two poplar species. Agrobacterium -mediated delivery of the shorter insert, TRV2 -PePDS 266 , into the host poplars resulted in expected photobleaching in both tree species, but not the longer insert, PePDS 558 . The efficiency of VIGS was temperature-dependent, increasing by raising the temperature from 18 to 28 °C. The optimized TRV–VIGS system at 28 °C resulted in a high silencing frequency and efficiency up to 65–73 and 83–94%, respectively, in the two tested poplars. Moreover, syringe inoculation of Agrobacterium in 100 mM acetosyringone induced a more efficient silencing in the two poplar species, compared with other agroinfiltration methods, e.g., direct injection, misting and agrodrench. There were plant species-related differences in the response to VIGS because the photobleaching symptoms were more severe in P.   x   canescens than in P. euphratica. Furthermore, VIGS-treated P. euphratica exhibited a higher recovery rate (50%) after several weeks of the virus infection, compared with TRV-infected P.   x   canescens plants (20%). Expression stability of reference genes was screened to assess the relative abundance of PePDS mRNA in VIGS-treated P. euphratica and P.   x   canescens. PeACT7 was stably expressed in P. euphratica and UBQ-L was selected as the most suitable reference gene for P.   x   canescens using three different statistical approaches, geNorm, NormFinder and BestKeeper. Quantitative real-time PCR showed significant reductions in PDS transcripts (55–64%) in the photobleached leaves of both VIGS-treated poplar species. Our results demonstrate that the TRV-based VIGS provides a practical tool for gene functional analysis in Populus sp., especially in those poplar species which are otherwise recalcitrant to transformation.

Description: Latex, the cytoplasm of laticiferous cells localized in the inner bark of rubber trees ( Hevea brasiliensis Müll. Arg.), is collected by tapping the bark. Following tapping, latex flows out of the trunk and is regenerated, whereas in untapped trees, there is no natural exudation. It is still unknown whether the carbohydrates used for latex regeneration in tapped trees is coming from recent photosynthates or from stored carbohydrates, and in the former case, it is expected that latex carbon isotope composition of tapped trees will vary seasonally, whereas latex isotope composition of untapped trees will be more stable. Temporal variations of carbon isotope composition of trunk latex ( 13 C-L), leaf soluble compounds ( 13 C-S) and bulk leaf material ( 13 C-B) collected from tapped and untapped 20-year-old trees were compared. A marked difference in 13 C-L was observed between tapped and untapped trees whatever the season. Trunk latex from tapped trees was more depleted (1.6 on average) with more variable 13 C values than those of untapped trees. 13 C-L was higher and more stable across seasons than 13 C-S and 13 C-B, with a maximum seasonal difference of 0.7 for tapped trees and 0.3 for untapped trees. 13 C-B was lower in tapped than in untapped trees, increasing from August (middle of the rainy season) to April (end of the dry season). Differences in 13 C-L and 13 C-B between tapped and untapped trees indicated that tapping affects the metabolism of both laticiferous cells and leaves. The lack of correlation between 13 C-L and 13 C-S suggests that recent photosynthates are mixed in the large pool of stored carbohydrates that are involved in latex regeneration after tapping.

Description: Trees contain non-structural carbon (NSC), but it is unclear for how long these reserves are stored and to what degree they are used to support plant activity. We used radiocarbon ( 14 C) to show that the carbon (C) in stemwood NSC can achieve ages of several decades in California oaks. We separated NSC into two fractions: soluble (~50% sugars) and insoluble (mostly starch) NSC. Soluble NSC contained more C than insoluble NSC, but we found no consistent trend in the amount of either pool with depth in the stem. There was no systematic difference in C age between the two fractions, although ages increased with stem depth. The C in both NSC fractions was consistently younger than the structural C from which they were extracted. Together, these results indicate considerable inward mixing of NSC within the stem and rapid exchange between soluble and insoluble pools, compared with the timescale of inward mixing. We observed similar patterns in sympatric evergreen and deciduous oaks and the largest differences among tree stems with different growth rates. The 14 C signature of carbon dioxide (CO 2 ) emitted from tree stems was higher than expected from very recent photoassimilates, indicating that the mean age of C in respiration substrates included a contribution from C fixed years previously. A simple model that tracks NSC produced each year, followed by loss (through conversion to CO 2 ) in subsequent years, matches our observations of inward mixing of NSC in the stem and higher 14 C signature of stem CO 2 efflux. Together, these data support the idea of continuous accumulation of NSC in stemwood and that ‘vigor’ (growth rate) and leaf habit (deciduous vs evergreen) control NSC pool size and allocation.

Description: Gibberellins (GAs) are important regulators of plant shoot biomass growth, and GA 20-oxidase (GA20ox) is one of the major regulatory enzymes in the GA biosynthetic pathway. Previously, we showed that the expression levels of a putative GA20ox1 (i.e., PdGA20ox1 ) in stem tissue of 3-month-old seedlings of 12 families of Pinus densiflora were positively correlated with stem diameter growth across those same families growing in an even-aged 32-year-old pine forest (Park EJ, Lee WY, Kurepin LV, Zhang R, Janzen L, Pharis RP (2015) Plant hormone-assisted early family selection in Pinus densiflora via a retrospective approach. Tree Physiol 35:86–94). To further investigate the molecular function of this gene in the stem wood growth of forest trees, we produced transgenic poplar lines expressing PdGA20ox1 under the control of the 35S promoter (designated as 35S::PdGA20ox1). By age 3 months, most of the 35S::PdGA20ox1 poplar trees were showing an exceptional enhancement of stem wood growth, i.e., up to fourfold increases in stem dry weight, compared with the nontransformed control poplar plants. Significant increases in endogenous GA 1 , its immediate precursor (GA 20 ) and its catabolite (GA 8 ) in elongating internode tissue accompanied the increased stem growth in the transgenic lines. Additionally, the development of gelatinous fibers occurred in vertically grown stems of the 35S::PdGA20ox1 poplars. An analysis of the cell wall monosaccharide composition of the 35S::PdGA20ox1 poplars showed significant increases in xylose and glucose contents, indicating a qualitative increase in secondary wall depositions. Microarray analyses led us to find a total of 276 probe sets that were upregulated (using threefold as a threshold) in the stem tissues of 35S::PdGA20ox1 poplars relative to the controls. ‘Cell organization or biogenesis’- and ‘cell wall’-related genes were overrepresented, including many of genes that are involved in cell wall modification. Several transcriptional regulators, which positively regulate cell elongation through GA signaling, were also upregulated. In contrast, genes involved in defense signaling were appreciably downregulated in the 35S::PdGA20ox1 stem tissues, suggesting a growth versus defense trade-off. Taken together, our results suggest that PdGA20ox1 functions to promote stem growth and wood formation in poplar, probably by activating GA signaling while coincidentally depressing defense signaling.

Description: Bark beetles (Coleoptera: Curculionidae, Scolytinae) cause widespread tree mortality in coniferous forests worldwide. Constitutive and induced host defenses are important factors in an individual tree’s ability to survive an attack and in bottom-up regulation of bark beetle population dynamics, yet quantifying defense levels is often difficult. For example, in Pinus spp., resin flow is important for resistance to bark beetles but is extremely variable among individuals and within a season. While resin is produced and stored in resin ducts, the specific resin duct metrics that best correlate with resin flow remain unclear. The ability and timing of some pine species to produce induced resin is also not well understood. We investigated (i) the relationships between ponderosa pine ( Pinus ponderosa Lawson & C. Lawson) resin flow and axial resin duct characteristics, tree growth and physiological variables, and (ii) if mechanical wounding induces ponderosa pine resin flow and resin ducts in the absence of bark beetles. Resin flow increased later in the growing season under moderate water stress and was highest in faster growing trees. The best predictors of resin flow were nonstandardized measures of resin ducts, resin duct size and total resin duct area, both of which increased with tree growth. However, while faster growing trees tended to produce more resin, models of resin flow using only tree growth were not statistically significant. Further, the standardized measures of resin ducts, density and duct area relative to xylem area, decreased with tree growth rate, indicating that slower growing trees invested more in resin duct defenses per unit area of radial growth, despite a tendency to produce less resin overall. We also found that mechanical wounding induced ponderosa pine defenses, but this response was slow. Resin flow increased after 28 days, and resin duct production did not increase until the following year. These slow induced responses may allow unsuccessfully attacked or wounded trees to resist future bark beetle attacks. Forest management that encourages healthy, vigorously growing trees will also favor larger resin ducts, thereby conferring increased constitutive resistance to bark beetle attacks.

Description: Temperature responses and sensitivity of photosynthesis ( A n _ T ) and respiration for leaves at different ages are crucial to modeling ecosystem carbon (C) cycles and productivity of evergreen forests. Understanding the mechanisms and processes of temperature sensitivity may further shed lights on temperature acclimation of photosynthesis and respiration with leaf aging. The current study examined temperature responses of photosynthesis and respiration of young leaves (YLs) (fully expanded in current growth season) and old leaves (OLs) (fully expanded in last growth season) of Quercus aquifolioides Rehder and E.H. Wilson in an alpine oak forest, southwestern China. Temperature responses of dark respiration ( R dark ), net assimilation ( A n ), maximal velocity of carboxylation ( V cmax ) and maximum rate of electron transport ( J max ) were significantly different between the two leaf ages. Those differences implied different temperature response parameters should be used for leaves of different ages in modeling vegetation productivity and ecosystem C cycles in Q. aquifolioides forests and other evergreen forests. We found that RuBP carboxylation determined the downward shift of A n _ T in OLs, while RuBP regeneration and the balance between Rubisco carboxylation and RuBP regeneration made little contribution. Sensitivity of stomatal conductance to vapor pressure deficit changed in OLs and compensated part of the downward shift. We also found that OLs of Q. aquifolioides had lower A n due to lower stomatal conductance, higher stomatal conductance limitation and deactivation of the biochemical processes. In addition, the balance between R dark and A n changed between OLs and YLs, which was represented by a higher R dark / A n ratio for OLs.

Description: Plants allocate carbon (C) to sink tissues depending on phenological, physiological or environmental factors. We still have little knowledge on C partitioning into various cellular compounds and metabolic pathways at various ecophysiological stages. We used compound-specific stable isotope analysis to investigate C partitioning of freshly assimilated C into tree compartments (needles, branches and stem) as well as into needle water-soluble organic C (WSOC), non-hydrolysable structural organic C (stOC) and individual chemical compound classes (amino acids, hemicellulose sugars, fatty acids and alkanes) of Norway spruce ( Picea abies ) following in situ 13 C pulse labelling 15 days after bud break. The 13 C allocation within the above-ground tree biomass demonstrated needles as a major C sink, accounting for 86% of the freshly assimilated C 6 h after labelling. In needles, the highest allocation occurred not only into the WSOC pool (44.1% of recovered needle 13 C) but also into stOC (33.9%). Needle growth, however, also caused high 13 C allocation into pathways not involved in the formation of structural compounds: (i) pathways in secondary metabolism, (ii) C-1 metabolism and (iii) amino acid synthesis from photorespiration. These pathways could be identified by a high 13 C enrichment of their key amino acids. In addition, 13 C was strongly allocated into the n -alkyl lipid fraction (0.3% of recovered 13 C), whereby 13 C allocation into cellular and cuticular exceeded that of epicuticular fatty acids. 13 C allocation decreased along the lipid transformation and translocation pathways: the allocation was highest for precursor fatty acids, lower for elongated fatty acids and lowest for the decarbonylated n -alkanes. The combination of 13 C pulse labelling with compound-specific 13 C analysis of key metabolites enabled tracing relevant C allocation pathways under field conditions. Besides the primary metabolism synthesizing structural cell compounds, a complex network of pathways consumed the assimilated 13 C and kept most of the assimilated C in the growing needles.

Description: Non-structural carbohydrates (NSC) in plant tissue are frequently quantified to make inferences about plant responses to environmental conditions. Laboratories publishing estimates of NSC of woody plants use many different methods to evaluate NSC. We asked whether NSC estimates in the recent literature could be quantitatively compared among studies. We also asked whether any differences among laboratories were related to the extraction and quantification methods used to determine starch and sugar concentrations. These questions were addressed by sending sub-samples collected from five woody plant tissues, which varied in NSC content and chemical composition, to 29 laboratories. Each laboratory analyzed the samples with their laboratory-specific protocols, based on recent publications, to determine concentrations of soluble sugars, starch and their sum, total NSC. Laboratory estimates differed substantially for all samples. For example, estimates for Eucalyptus globulus leaves (EGL) varied from 23 to 116 (mean = 56) mg g –1 for soluble sugars, 6–533 (mean = 94) mg g –1 for starch and 53–649 (mean = 153) mg g –1 for total NSC. Mixed model analysis of variance showed that much of the variability among laboratories was unrelated to the categories we used for extraction and quantification methods (method category R 2 = 0.05–0.12 for soluble sugars, 0.10–0.33 for starch and 0.01–0.09 for total NSC). For EGL, the difference between the highest and lowest least squares means for categories in the mixed model analysis was 33 mg g –1 for total NSC, compared with the range of laboratory estimates of 596 mg g –1 . Laboratories were reasonably consistent in their ranks of estimates among tissues for starch ( r = 0.41–0.91), but less so for total NSC ( r = 0.45–0.84) and soluble sugars ( r = 0.11–0.83). Our results show that NSC estimates for woody plant tissues cannot be compared among laboratories. The relative changes in NSC between treatments measured within a laboratory may be comparable within and between laboratories, especially for starch. To obtain comparable NSC estimates, we suggest that users can either adopt the reference method given in this publication, or report estimates for a portion of samples using the reference method, and report estimates for a standard reference material. Researchers interested in NSC estimates should work to identify and adopt standard methods.

Description: Several hypotheses exist that describe phytoplankton spring blooms in temperate and subpolar oceans: the critical depth, shoaling mixed layer (ML), critical turbulence, onset of stratification and disturbance-recovery hypotheses. These theories appear to be mutually exclusive and none of them describe the annual cycle of phytoplankton biomass. Here, we present a model of the annual cycle in phytoplankton that recognizes that phytoplankton are not always mixed throughout the so-called ML, and that it is important to distinguish between the surface biomass and depth-integrated phytoplankton. Once these important distinctions are made, the annual cycles and blooms in surface and depth-integrated phytoplankton can be described straightforwardly in terms of the physical drivers and biotic responses.

Description: Impacts of climate change on marine ecosystems have become increasingly apparent during the past decades. In consequence, it is necessary to study how these alterations can affect the habitat and population dynamics of key organisms. Here we used a video plankton recorder (VPR) to investigate the effect of climate-induced habitat changes on the copepod Pseudocalanus acuspes , a key species in the Baltic Sea. The VPR allowed the observation of reproducing copepod females, identified by attached egg sacs, usually lost during traditional net sampling. We compared the small-scale distribution of our target species during non-inflow and inflow periods. Our study showed a large increase in the availability of suitable habitat after the inflow event due to improved oxygen and salinity conditions. Furthermore, increased copepod abundance and a deeper and wider vertical distribution was apparent. Applying a new approach to estimate in situ egg production rates from VPR-derived images revealed no changes. However, we observed increased offspring survival with improved hydrographic conditions pointing toward the importance of salinity and oxygen for the population dynamics of Baltic P. acuspes . Our observations illustrate the strong impact that climate change can have on the habitat of key marine ecosystem species, important for overall ecosystem dynamics.

Description: Research on nutrient controls of planktonic productivity tends to focus on a few standard fractions of inorganic or total nitrogen (N) and phosphorus (P). However, there is a wide range in the degree to which land-derived dissolved organic nutrients can be assimilated by biota. Thus, in systems where such fractions form a majority of the macronutrient resource pool, including many boreal inland waters and estuaries, our understanding of bacterio- and phytoplankton production dynamics remains limited. To adequately predict aquatic productivity in a changing environment, improved standard methods are needed for determining the sizes of active (bioavailable) pools of N, P and organic carbon (C). A synthesis of current knowledge suggests that variation in the C:N:P stoichiometry of bioavailable resources is associated with diverse processes that differentially influence the individual elements across space and time. Due to a generally increasing organic nutrient bioavailability from C to N to P, we hypothesize that the C:N and N:P of bulk resources often vastly overestimates the corresponding ratios of bioavailable resources. It is further proposed that basal planktonic production is regulated by variation in the source, magnitude and timing of terrestrial runoff, through processes that have so far been poorly described.

Description: Understanding the factors that regulate the abundance, size structure and community structure of cladocerans is an important goal of aquatic ecologists. While both top-down and bottom-up factors help to structure cladoceran communities, there may be interactions between these factors. We conducted a mesocosm study to determine how alien large-bodied Daphnia , zebra mussels and fish affected cladoceran community and size structure. We found that large-bodied Daphnia reduced algal resources and the fecundity of smaller bodied cladocerans. Fish removed the large-bodied Daphnia magna from the mesocosms and shifted the cladoceran community to a smaller body size. Fish also appeared to promote increases in cladoceran diversity through the coexistence of several smaller bodied taxa. In contrast, zebra mussels increased cyanobacteria and helped to promote the success of the alien Daphnia , but reduced the biomass of small-bodied cladocerans. Zebra mussels reduced the carbon (C):phosphorus (P) ratio of the phytoplankton in the mesocosms which may have favored the relatively P-limited Daphnia . Combined, our results highlight the complex interactions of multiple factors that help to regulate cladoceran community and size structure.

Description: In freshwater systems, Daphnia has been demonstrated to show adaptive responses following the light–dark cycle. The adjustment of these responses to the change of day and night is probably transmitted via the hormone melatonin. The rate-limiting enzyme in melatonin synthesis is the arylalkylamine N-transferase (AANAT). We identified three genes coding for insect-like AANATs in Daphnia , of which we measured the gene expression in an ecologically relevant light–dark cycle. We demonstrated that Daphnia 's insect-like AANAT gene expression oscillated in a daily manner, and that the highest peak of expression after the onset of darkness was followed by a peak of melatonin production at midnight. Moreover, we could show an oscillation of endogenous melatonin synthesis in Daphnia . In most organisms, melatonin synthesis is due to rhythmic expression of genes of the circadian clock, since transcription of aanat s is directly linked to a circadian transcription factor. We could demonstrate that putative clock genes and insect-like AANAT genes of Daphnia were equally expressed. Therefore, we propose that melatonin synthesis is coupled to the expression of Daphnia clock genes, and that insect-like AANATs of crustaceans have a similar function as AANATs of vertebrates: The initiation of melatonin synthesis. In future studies with Daphnia , it will be necessary to take the time of day into account since melatonin concentrations might influence stress responses.

Description: It has been hypothesized that terrestrial particulate organic matter (t-POM) makes important contributions to Daphnia production in some lakes. We conducted a series of feeding experiments to explore the fatty acid responses in Daphnia to diets comprised of different terrestrial resources (i.e. Alnus rubra , Phragmites australis , Betula nana and Betula pendula ) and mixed diets with terrestrial and phytoplankton ( Scenedesmus or Cryptomonas ) resources. When fed 100% phytoplankton, Daphnia had very similar ( r 2 〉 0.80) fatty acid profiles to their diets, whereas Daphnia that consumed t-POM diets had weak correlations ( r 2 = 0.002–0.56) with the corresponding diet sources. Unusual 16 carbon chain polyunsaturated fatty acids (16:26, 16:33 and 16:43), linoleic acid (18:26) and α-linolenic acid (18:33) were diagnostic fatty acids for Scenedesmus and Daphnia that consumed this alga. Stearidonic acid (18:43) and eicosapentaenoic acid (20:53) were diagnostic for Cryptomonas and Daphnia that consumed this diet. All of the t-POM resources were characterized by a high content of saturated fatty acids (SAFA; 79 ± 12%), especially the diagnostic long-chain SAFA (20:0, 22:0, 24:0, 26:0, 28:0). Daphnia that consumed t-POM assimilated very little of these terrestrial biomarkers, but the shorter chain SAFA 16:0 and 18:0 were very prevalent in juvenile and adult Daphnia that consumed terrestrial plant matter. The -3:-6 ratios were distinctive between terrestrial (0.3–1.6) and phytoplankton resources (3–15), and this ratio in Daphnia was strongly associated with their diets ( r 2 = 0.88). These results suggest that Daphnia , and perhaps zooplankton in general, preferentially retain algae-derived 3 fatty acids, and low -3:-6 ratios in Daphnia indicate a mainly terrestrial diet or poor nutritional condition.

Description: In order to trace community dynamics and reticulate evolution in hybrid species complexes, long-term comparative studies of natural populations are necessary. Such studies require the development of tools for fine-scale genetic analyses. In the present study, we developed species-diagnostic SNP-based markers for hybridizing freshwater crustaceans: the multispecies Daphnia longispina complex. Specifically, we took advantage of transcriptome data from a key species of this hybrid complex, the annotated genome of a related Daphnia species and well-defined reference genotypes from three parental species. Altogether eleven nuclear loci with several species-specific SNP sites were identified in sequence alignments of these reference genotypes from three parental species and their interspecific hybrids. A PCR-RFLP assay was developed for cost-efficient large population screening by SNP-based genotyping. Taxon assignment by RFLP patterns was nearly perfectly concordant with microsatellite genotyping across several screened populations from Europe. Finally, we were able to amplify two short regions of these loci in formaldehyde-preserved samples dating back to the year 1960. The species-specific SNP-based markers developed here provide valuable tools to study hybridization over time, including the long-term impact of various environmental factors on hybridization and biodiversity changes. SNP-based genotyping will finally allow eco-evolutionary dynamics to be revealed at different time scales.

Description: We employed the warm temperate conifer Cunninghamia lanceolata (Lamb.) Hook. as a model of plantation forest species to investigate ecophysiological responses to root treatments (control (0%), and ~25, 50 or 75% of the initial root mass) under well-watered and water-limited conditions. Our results indicated that total root dry mass accumulation was negatively associated with the severity of root pruning, but there was evidence of multiple compensatory responses. The plants exhibited higher instantaneous and long-term (assessed by carbon isotope composition, 13 C) water-use efficiency in pruning treatments, especially under low water availability. Root pruning also increased the fine root/total root mass ratio, specific root length and fine root vitality in both water availability treatments. As a result of the compensatory responses, under well-watered conditions, height, stem dry mass accumulation, leaf/fine root biomass ratio (L/FR), transpiration rate, photosynthetic capacity and photosynthetic nitrogen-use efficiency ( E N ) were the highest under 25% pruning. Yet, all these traits except L/FR and foliage nitrogen content were severely reduced under 75% pruning. Drought negatively affected growth and leaf gas exchange rates, and there was a greater negative effect on growth, water potential, gas exchange and E N when 〉25% of total root biomass was removed. The stem/aboveground mass ratio was the highest under 25% pruning in both watering conditions. These results indicate that the responses to root severance are related to the excision intensity and soil moisture content. A moderate root pruning proved to be an effective means to improve stem dry mass accumulation.

Description: The timing of wood formation is crucial to determine how environmental factors affect tree growth. The long-lived bristlecone pine ( Pinus longaeva D. K. Bailey) is a foundation treeline species in the Great Basin of North America reaching stem ages of about 5000 years. We investigated stem cambial phenology and radial size variability to quantify the relative influence of environmental variables on bristlecone pine growth. Repeated cellular measurements and half-hourly dendrometer records were obtained during 2013 and 2014 for two high-elevation stands included in the Nevada Climate-ecohydrological Assessment Network. Daily time series of stem radial variations showed rehydration and expansion starting in late April–early May, prior to the onset of wood formation at breast height. Formation of new xylem started in June and lasted until mid-September. There were no differences in phenological timing between the two stands, or in the air and soil temperature thresholds for the onset of xylogenesis. A multiple logistic regression model highlighted a separate effect of air and soil temperature on xylogenesis, the relevance of which was modulated by the interaction with vapor pressure and soil water content. While air temperature plays a key role in cambial resumption after winter dormancy, soil thermal conditions coupled with snowpack dynamics also influence the onset of wood formation by regulating plant–soil water exchanges. Our results help build a physiological understanding of climate–growth relationships in P. longaeva , the importance of which for dendroclimatic reconstructions can hardly be overstated. In addition, environmental drivers of xylogenesis at the treeline ecotone, by controlling the growth of dominant species, ultimately determine ecosystem responses to climatic change.

Description: Seasonal analyses of cambial cell production and day-by-day stem radial increment can help to elucidate how climate modulates wood formation in conifers. Intra-annual dynamics of wood formation were determined with microcores and dendrometers and related to climatic signals in Norway spruce ( Picea abies (L.) Karst.). The seasonal dynamics of these processes were observed at two sites of different altitude, Savignano (650 m a.s.l.) and Lavazè (1800 m a.s.l.) in the Italian Alps. Seasonal dynamics of cambial activity were found to be site specific, indicating that the phenology of cambial cell production is highly variable and plastic with altitude. There was a site-specific trend in the number of cells in the wall thickening phase, with the maximum cell production in early July (DOY 186) at Savignano and in mid-July (DOY 200) at Lavazè. The formation of mature cells showed similar trends at the two sites, although different numbers of cells and timing of cell differentiation were visible in the model shapes; at the end of ring formation in 2010, the number of cells was four times higher at Savignano (106.5 cells) than at Lavazè (26.5 cells). At low altitudes, microcores and dendrometers described the radial growth patterns comparably, though the dendrometer function underlined the higher upper asymptote of maximum growth in comparison with the cell production function. In contrast, at high altitude, these functions exhibited different trends. The best model was obtained by fitting functions of the Gompertz model to the experimental data. By combining radial growth and cambial activity indices we defined a model system able to synchronize these processes. Processes of adaptation of the pattern of xylogenesis occurred, enabling P. abies to occupy sites with contrasting climatic conditions. The use of daily climatic variables in combination with plant functional traits obtained by sensors and/or destructive sampling could provide a suitable tool to better investigate the effect of disturbances on response strategies in trees and, consequently, contribute to improving our prediction of tree growth and species resilience based on climate scenarios.

Description: In deciduous trees growing in temperate forests, bud break and growth in spring must rely on intrinsic carbon (C) reserves. Yet it is unclear whether growth and C storage occur simultaneously, and whether starch C in branches is sufficient for refoliation. To test in situ the relationships between growth, phenology and C utilization, we monitored stem growth, leaf phenology and stem and branch nonstructural carbohydrate (NSC) dynamics in three deciduous species: Carpinus betulus L., Fagus sylvatica L. and Quercus petraea (Matt.) Liebl. To quantify the role of NSC in C investment into growth, a C balance approach was applied. Across the three species, 〉95% of branchlet starch was consumed during bud break, confirming the importance of C reserves for refoliation in spring. The C balance calculation showed that 90% of the C investment in foliage (7.0–10.5 kg tree –1 and 5–17 times the C needed for annual stem growth) was explained by simultaneous branchlet starch degradation. Carbon reserves were recovered sooner than expected, after leaf expansion, in parallel with stem growth. Carpinus had earlier leaf phenology (by ~25 days) but delayed cambial growth (by ~15 days) than Fagus and Quercus , the result of a competitive strategy to flush early, while having lower NSC levels.

Description: Fungal infections result in decreases in photosynthesis, induction of stress and signaling volatile emissions and reductions in constitutive volatile emissions, but the way different physiological processes scale with the severity of infection is poorly known. We studied the effects of infection by the obligate biotrophic fungal pathogen Melampsora larici-populina Kleb., the causal agent of poplar leaf rust disease, on photosynthetic characteristics, and constitutive isoprene and induced volatile emissions in leaves of Populus balsamifera var. suaveolens (Fisch.) Loudon. exhibiting different degrees of damage. The degree of fungal damage, quantified by the total area of chlorotic and necrotic leaf areas, varied between 0 (noninfected control) and ~60%. The rates of all physiological processes scaled quantitatively with the degree of visual damage, but the scaling with damage severity was weaker for photosynthetic characteristics than for constitutive and induced volatile release. Over the whole range of damage severity, the net assimilation rate per area ( A A ) decreased 1.5-fold, dry mass per unit area 2.4-fold and constitutive isoprene emissions 5-fold, while stomatal conductance increased 1.9-fold and dark respiration rate 1.6-fold. The emissions of key stress and signaling volatiles (methanol, green leaf volatiles, monoterpenes, sesquiterpenes and methyl salicylate) were in most cases nondetectable in noninfested leaves, and increased strongly with increasing the spread of infection. The moderate reduction in A A resulted from the loss of photosynthetically active biomass, but the reduction in constitutive isoprene emissions and the increase in induced volatile emissions primarily reflected changes in the activities of corresponding biochemical pathways. Although all physiological alterations in fungal-infected leaves occurred in a stress severity-dependent manner, modifications in primary and secondary metabolic pathways scaled differently due to contrasting operational mechanisms.

Description: Current knowledge of the genetic mechanisms underlying the inheritance of photosynthetic activity in forest trees is generally limited, yet it is essential both for various practical forestry purposes and for better understanding of broader evolutionary mechanisms. In this study, we investigated genetic variation underlying selected chlorophyll a fluorescence (ChlF) parameters in structured populations of Scots pine ( Pinus sylvestris L.) grown on two sites under non-stress conditions. These parameters were derived from the OJIP part of the ChlF kinetics curve and characterize individual parts of primary photosynthetic processes associated, for example, with the exciton trapping by light-harvesting antennae, energy utilization in photosystem II (PSII) reaction centers (RCs) and its transfer further down the photosynthetic electron-transport chain. An additive relationship matrix was estimated based on pedigree reconstruction, utilizing a set of highly polymorphic single sequence repeat markers. Variance decomposition was conducted using the animal genetic evaluation mixed-linear model. The majority of ChlF parameters in the analyzed pine populations showed significant additive genetic variation. Statistically significant heritability estimates were obtained for most ChlF indices, with the exception of DI 0 /RC, D0 and P0 ( F v / F m ) parameters. Estimated heritabilities varied around the value of 0.15 with the maximal value of 0.23 in the ET 0 /RC parameter, which indicates electron-transport flux from Q A to Q B per PSII RC. No significant correlation was found between these indices and selected growth traits. Moreover, no genotype  x  environment interaction (G  x  E) was detected, i.e., no differences in genotypes’ performance between sites. The absence of significant G  x  E in our study is interesting, given the relatively low heritability found for the majority of parameters analyzed. Therefore, we infer that polygenic variability of these indices is selectively neutral.

Description: The ethylene response factor (ERF) family is one of the largest plant-specific transcription factor families, playing an important role in plant development and response to stresses. The ERF76 gene is a member of the poplar ERF transcription factor gene family. First, we validated that the ERF76 gene expressed in leaf and root tissues is responsive to salinity stress. We then successfully cloned the ERF76 cDNA fragment containing an open reading frame from di-haploid Populus simonii   x   Populus nigra and proved that ERF76 protein is targeted to the nucleus. Finally, we transferred the gene into the same poplar clone by the Agrobacterium -mediated leaf disc method. Using both RNA-Seq and reverse transcription-quantitative polymerase chain reaction, we validated that expression level of ERF76 is significantly higher in transgenic plants than that in the nontransgenic control. Using RNA-Seq data, we have identified 375 genes that are differentially expressed between the transgenic plants and the control under salt treatment. Among the differentially expressed genes, 16 are transcription factor genes and 45 are stress-related genes, both of which are upregulated significantly in transgenic plants, compared with the control. Under salt stress, the transgenic plants showed significant increases in plant height, root length, fresh weight, and abscisic acid (ABA) and gibberellin (GA) concentration compared with the control, suggesting that overexpression of ERF76 in transgenic poplar upregulated the expression of stress-related genes and increased the ability of ABA and GA biosynthesis, which resulted in stronger tolerance to salt stress.

Description: Summer droughts are likely to increase in frequency and intensity across Europe, yet long-lived trees may have a limited ability to tolerate drought. It is therefore critical that we improve our understanding of phenotypic plasticity to drought in natural populations for ecologically and economically important trees such as Populus nigra L. A common garden experiment was conducted using ~500 wild P. nigra trees, collected from 11 river populations across Europe. Phenotypic variation was found across the collection, with southern genotypes from Spain and France characterized by small leaves and limited biomass production. To examine the relationship between phenotypic variation and drought tolerance, six genotypes with contrasting leaf morphologies were subjected to a water deficit experiment. ‘North eastern’ genotypes were collected at wet sites and responded to water deficit with reduced biomass growth, slow stomatal closure and reduced water use efficiency (WUE) assessed by 13 C. In contrast, ‘southern’ genotypes originating from arid sites showed rapid stomatal closure, improved WUE and limited leaf loss. Transcriptome analyses of a genotype from Spain (Sp2, originating from an arid site) and another from northern Italy (Ita, originating from a wet site) revealed dramatic differences in gene expression response to water deficit. Transcripts controlling leaf development and stomatal patterning, including SPCH , ANT , ER , AS1 , AS2 , PHB , CLV1 , ERL1–3 and TMM , were down-regulated in Ita but not in Sp2 in response to drought.

Description: Isoprene is the most abundant type of nonmethane, biogenic volatile organic compound in the atmosphere, and it is produced mainly by terrestrial plants. The tropical tree species Ficus septica Burm. F. (Rosales: Moraceae) has been shown to cease isoprene emissions when exposed to temperatures of 12 °C or lower and to re-induce isoprene synthesis upon subsequent exposure to temperatures of 30 °C or higher for 24 h. To elucidate the regulation of genes underlying the disabling and then induction of isoprene emission during acclimatization to ambient temperature, we conducted gene expression analyses of F. septica plants under changing temperature using quantitative real-time polymerase chain reaction and western blotting. Transcription levels were analyzed for 17 genes that are involved in metabolic pathways potentially associated with isoprene biosynthesis, including isoprene synthase ( ispS ). The protein levels of ispS were also measured. Changes in transcription and protein levels of the ispS gene, but not in the other assessed genes, showed identical temporal patterns to isoprene emission capacity under the changing temperature regime. The ispS protein levels strongly and positively correlated with isoprene emission capacity ( R 2  = 0.92). These results suggest that transcriptional regulation of ispS gave rise to the temporal variation in isoprene emission capacity in response to changing temperature.

Description: Clonal integration between ramets can be an ecological advantage of clonal plant species in environments where resources are patchily distributed. We investigated physiological integration among Populus balsamifera L. ramets under drought stress in order to demonstrate water sharing between connected ramets. Pairs of connected ramets were grown in separate pots in the greenhouse where half of ramets had the parental root connection severed and half were left intact. Drought stress was applied to one ramet, and growth, specific leaf area (SLA), net photosynthesis, stomatal conductance, leaf water potential and carbon isotopic composition ( 13 C) were measured after an 8-week growing period. Droughted ramets connected to watered ramets were able to maintain high gas exchange activity and water potential, similar to watered ramets. Leaf water potential and SLA results showed that the root connection was more beneficial for proximal compared with distal ramets. The parental root connection also allowed droughted ramets to discriminate more against 13 C compared with severed ramets. In conclusion, this study shows compelling evidence of physiological integration of connected P. balsamifera ramets through water sharing.

Description: Wood biophysical properties and the dynamics of water storage discharge and refilling were studied in the trunk of canopy tree species with diverse life history and functional traits in subtropical forests of northeast Argentina. Multiple techniques assessing capacitance and storage capacity were used simultaneously to improve our understanding of the functional significance of internal water sources in trunks of large trees. Sapwood capacitances of 10 tree species were characterized using pressure–volume relationships of sapwood samples obtained from the trunk. Frequency domain reflectometry was used to continuously monitor the volumetric water content in the main stems. Simultaneous sap flow measurements on branches and at the base of the tree trunk, as well as diurnal variations in trunk contraction and expansion, were used as additional measures of stem water storage use and refilling dynamics. All evidence indicates that tree trunk internal water storage contributes from 6 to 28% of the daily water budget of large trees depending on the species. The contribution of stored water in stems of trees to total daily transpiration was greater for deciduous species, which exhibited higher capacitance and lower sapwood density. A linear relationship across species was observed between wood density and growth rates with the higher wood density species (mostly evergreen) associated with lower growth rates and the lower wood density species (mostly deciduous) associated with higher growth rates. The large sapwood capacitance in deciduous species may help to avoid catastrophic embolism in xylem conduits. This may be a low-cost adaptation to avoid water deficits during peak water use at midday and under temporary drought periods and will contribute to higher growth rates in deciduous tree species compared with evergreen ones. Large capacitance appears to have a central role in the rapid growth patterns of deciduous species facilitating rapid canopy access as these species are less shade tolerant than evergreen species.

Description: Nuclear magnetic resonance (NMR) and NMR imaging (magnetic resonance imaging) offer the possibility to quantitatively and non-invasively measure the presence and movement of water. Unfortunately, traditional NMR hardware is expensive, poorly suited for plants, and because of its bulk and complexity, not suitable for use in the field. But does it need to be? We here explore how novel, small-scale portable NMR devices can be used as a flow sensor to directly measure xylem sap flow in a poplar tree ( Populus nigra L.), or in a dendrometer-like fashion to measure dynamic changes in the absolute water content of fruit or stems. For the latter purpose we monitored the diurnal pattern of growth, expansion and shrinkage in a model fruit (bean pod, Phaseolus vulgaris L.) and in the stem of an oak tree ( Quercus robur L.). We compared changes in absolute stem water content, as measured by the NMR sensor, against stem diameter variations as measured by a set of conventional point dendrometers, to test how well the sensitivities of the two methods compare and to investigate how well diurnal changes in trunk absolute water content correlate with the concomitant diurnal variations in stem diameter. Our results confirm the existence of a strong correlation between the two parameters, but also suggest that dynamic changes in oak stem water content could be larger than is apparent on the basis of the stem diameter variation alone.

Description: While natural spatial temperature gradients between measurement needles have been thoroughly investigated for continuous heat-based sap flow methods, little attention has been given to how natural changes in stem temperature impact heat pulse-based methods through temporal rather than spatial effects. By modelling the theoretical equation for both an ideal instantaneous pulse and a step pulse and applying a finite element model which included actual needle dimensions and wound effects, the influence of a varying stem temperature on heat pulse-based methods was investigated. It was shown that the heat ratio (HR) method was influenced, while for the compensation heat pulse and T max methods changes in stem temperatures of up to 0.002 °C s –1 did not lead to significantly different results. For the HR method, rising stem temperatures during measurements led to lower heat pulse velocity values, while decreasing stem temperatures led to both higher and lower heat pulse velocities, and to imaginary results for high flows. These errors of up to 40% can easily be prevented by including a temperature correction in the data analysis procedure, calculating the slope of the natural temperature change based on the measured temperatures before application of the heat pulse. Results of a greenhouse and outdoor experiment on Pinus pinea L. show the influence of this correction on low and average sap flux densities.

Description: The control of plant transpiration by stomata under water stress and recovery conditions is of paramount importance for plant performance and survival. Although both chemical and hydraulic signals emitted within a plant are considered to play a major role in controlling stomatal dynamics, they have rarely been assessed together. The aims of this study were to evaluate (i) the dynamics of chemical and hydraulic signals at leaf, stem and root level, and (ii) their effect on the regulation of stomatal conductance ( g s ) during water stress and recovery. Measurements of g s , water potential, abscisic acid (ABA) content and loss of hydraulic functioning at leaf, stem and root level were conducted during a water stress and recovery period imposed on 1-year-old olive plants ( Olea europaea L.). Results showed a strong hydraulic segmentation in olive plants, with higher hydraulic functioning losses in roots and leaves than in stems. The dynamics of hydraulic conductance of roots and leaves observed as water stress developed could explain both a protection of the hydraulic functionality of larger organs of the plant (i.e., branches, etc.) and a role in the down-regulation of g s . On the other hand, ABA also increased, showing a similar pattern to g s dynamics, and thus its effect on g s in response to water stress cannot be ruled out. However, neither hydraulic nor non-hydraulic factors were able to explain the delay in the full recovery of g s after soil water availability was restored.

Description: For isohydric trees mid-day water uptake is stable and depends on soil water status, reflected in pre-dawn leaf water potential ( pd ) and mid-day stem water potential ( md ), tree hydraulic conductance and a more-or-less constant leaf water potential ( l ) for much of the day, maintained by the stomata. Stabilization of l can be represented by a linear relationship between canopy resistance ( R c ) and vapor pressure deficit ( D ), and the slope ( B D ) is proportional to the steady-state water uptake. By analyzing sap flow (SF), meteorological and md measurements during a series of wetting and drying ( D / W ) cycles in a nectarine orchard, we found that for the range of md relevant for irrigated orchards the slope of the relationship of R c to D , B D is a linear function of md . R c was simulated using the above relationships, and its changes in the morning and evening were simulated using a rectangular hyperbolic relationship between leaf conductance and photosynthetic irradiance, fitted to leaf-level measurements. The latter was integrated with one-leaf, two-leaf and integrative radiation models, and the latter gave the best results. Simulated R c was used in the Penman–Monteith equation to simulate tree transpiration, which was validated by comparing with SF from a separate data set. The model gave accurate estimates of diurnal and daily total tree transpiration for the range of md s used in regular and deficit irrigation. Diurnal changes in tree water content were determined from the difference between simulated transpiration and measured SF . Changes in water content caused a time lag of 90–105 min between transpiration and SF for md between –0.8 and –1.55 MPa, and water depletion reached 3 l h –1 before noon. Estimated mean diurnal changes in water content were 5.5 l day –1  tree –1 at md of –0.9 MPa and increased to 12.5 l day –1  tree –1 at –1.45 MPa, equivalent to 6.5 and 16.5% of daily tree water use, respectively. Sixteen percent of the dynamic water volume was in the leaves. Inversion of the model shows that md can be predicted from D and R c , which may have some importance for irrigation management to maintain target values of md . That relationship will be explored in future research.

Description: Prey size selectivity in piscivorous fish larvae is important to both aquaculture and fisheries science, but laboratory experiments are few. We analyzed selective foraging in Atlantic bluefin tuna larvae ( Thunnus thynnus ) using two larval fish prey species. The experiments revealed that selective foraging of prey sizes differed among bluefin tuna predator sizes (15–25 mm SL) and prey species, bonito ( Sarda sarda ) and seabream ( Sparus aurata ). The observed pattern suggest a general preference for small bonito prey larvae but large seabream prey. Thus, prey size alone is not the only trait responsible for size selectivity in piscivorous fish larvae.

Description: Using a unique 50-year high-resolution time series of daily kom-fyke catches, long-term patterns of scyphomedusae in the western Dutch Wadden Sea were analysed and related to changes in environmental conditions [eutrophication in the 1980s–1990s and recent climate change (increased water temperature)] in the area. Over the years, species composition and general pattern of appearance has remained the same: the first species that occurred in spring was Aurelia aurita , followed by Cyanea lamarckii / C. capillata. Chrysaora hysoscella and Rhizostoma octopus occurred from June to July onwards. All species appeared earlier in recent decades and first appearance and peak occurrence of A. aurita was in part inversely related to previous winter seawater temperature. Last occurrence of C. hysoscella was related to summer seawater temperature and the species is present longer in recent decades. Phenological relationships might have been decoupled since the seasonality of the phytoplankton bloom did not change. All species showed large inter-annual abundance fluctuations, with prolific years followed by sparse years. Peak catches of the coastal species A. aurita occurred in the late 1970s–early 1990s when eutrophication peaked, however, without a significant relationship with total nitrogen input into the area. Unlike for phenology, the patterns of mean abundance of any species did not show a relationship to climate change in the area. This might imply that population regulating mechanisms do not operate during the planktonic phase but during the sessile demersal polyp stages.

Description: In situ fluorometers are the optimal means of providing high-frequency estimates of phytoplankton communities. However, they may be subjected to measurement biases originating from variations in the physiological states of cells, the use of spectral fluorescence signatures (SFS) defined on the basis of inappropriate phytoplankton groups and the lack of linear independence between selected sets of SFS. We assessed correction procedures for measurement biases in mono and mixed cultures of five freshwater phytoplankton species. We investigated the impacts of total Chl a levels, the lack of linear independence between SFS and varying physiological states on the accuracy of the Chl a estimates that were provided by the FluoroProbe (bbe Moldaenke GmbH, Germany). The use of species-specific SFS allowed for the correction of quantification and classification biases. In some cases, the procedure led to a lack of linear independence between SFS, which significantly reduced estimation accuracies. A convenient method to evaluate linear independence between SFS is provided. Differences in the physiological states of phytoplankton cultures following light pre-acclimation and/or N-starvation appeared to be species specific. Light pre-acclimation led to an underestimation of biomass (up to –28.5%) through fluorescence quenching. The responses of the phytoplankton cultures to N-starvation varied depending on the species (from –40.3 to +336% biases in Chl a quantification). Overall, the application of appropriate corrective measures increased data accuracy. However, optimal data reliability can only be achieved by estimating phytoplankton community composition and associated environmental conditions.

Description: Cyanobacteria are the primary taxa responsible for freshwater harmful algal blooms (HABs), with several genera capable of producing potent intracellular toxins and off-flavor compounds. There is considerable growing interest in methods to rapidly quantify cyanobacteria in water samples. Past studies have demonstrated poor correlations between phycocyanin in vivo fluorescence and cyanobacterial cell densities. We conducted a series of laboratory experiments aimed at refining a protocol that uses benchtop fluorometry to measure the cyanobacterial pigment, phycocyanin, to accurately estimate cyanobacterial biovolume. In our study, we found strong correlations between phycocyanin concentration and cyanobacterial biovolume (but not for cell densities) both within and across ponds, which varied widely in productivity and algal diversity. Thus, benchtop fluorometry of phycocyanin is a viable method for water resource managers to quickly estimate cyanobacterial biovolume.

Description: Several dinoflagellate species in the genus Blastodinium are gut parasites of marine planktonic copepods. However, there is only limited information on the occurrence and infection frequencies of Blastodinium spp. in the field and almost no information on the functional impact on their hosts. We report upon the effects of Blastodinium sp . infection on Calanus finmarchicus from the northeastern Atlantic coast off southern Norway during April 2013 and 2014. Up to 58% of C. finmarchicus were infected near the coast, while 〈5% were infected several kilometers offshore. Ingestion rates of infected females were below detection limits and significantly lower than uninfected females. Blastodinium sp . -infected females showed characteristic symptoms of starvation, including lower respiration rates (implying a lower metabolic rate), production of smaller and fewer fecal pellets and significantly fewer eggs than uninfected females. A few females in this study were able to void the infection, however the extended period of starvation is likely to have longer-term repercussions on egg production rates well after the copepod clears the infection. The degree to which the infection affects C. finmarchicus recruitment depends on the extent of the spatial distribution of the infection. Monitoring of parasitic infection during routine field surveys will be required in order to clarify this.

Description: Drought-related tree die-off episodes have been observed in all vegetated continents. Despite much research effort, however, the multiple interactions between carbon starvation, hydraulic failure and biotic agents in driving tree mortality under field conditions are still not well understood. We analysed the seasonal variability of non-structural carbohydrates (NSCs) in four organs (leaves, branches, trunk and roots), the vulnerability to embolism in roots and branches, native embolism (percentage loss of hydraulic conductivity (PLC)) in branches and the presence of root rot pathogens in defoliated and non-defoliated individuals in a declining Scots pine ( Pinus sylvestris L.) population in the NE Iberian Peninsula in 2012, which included a particularly dry and warm summer. No differences were observed between defoliated and non-defoliated pines in hydraulic parameters, except for a higher vulnerability to embolism at pressures below –2 MPa in roots of defoliated pines. No differences were found between defoliation classes in branch PLC. Total NSC (TNSC, soluble sugars plus starch) values decreased during drought, particularly in leaves. Defoliation reduced TNSC levels across tree organs, especially just before (June) and during (August) drought. Root rot infection by the fungal pathogen Onnia P. Karst spp. was detected but it did not appear to be associated to tree defoliation. However, Onnia infection was associated with reduced leaf-specific hydraulic conductivity and sapwood depth, and thus contributed to hydraulic impairment, especially in defoliated pines. Infection was also associated with virtually depleted root starch reserves during and after drought in defoliated pines. Moreover, defoliated and infected trees tended to show lower basal area increment. Overall, our results show the intertwined nature of physiological mechanisms leading to drought-induced mortality and the inherent difficulty of isolating their contribution under field conditions.

Description: Non-structural carbohydrates (NSCs) are critical to maintain plant metabolism under stressful environmental conditions, but we do not fully understand how NSC allocation and utilization from storage varies with stress. While it has become established that storage allocation is unlikely to be a mere overflow process, very little empirical evidence has been produced to support this view, at least not for trees. Here we present the results of an intensively monitored experimental manipulation of whole-tree carbon (C) balance (young Picea abies (L.) H Karst.) using reduced atmospheric [CO 2 ] and drought to reduce C sources. We measured specific C storage pools (glucose, fructose, sucrose, starch) over 21 weeks and converted concentration measurement into fluxes into and out of the storage pool. Continuous labeling ( 13 C) allowed us to track C allocation to biomass and non-structural C pools. Net C fluxes into the storage pool occurred mainly when the C balance was positive. Storage pools increased during periods of positive C gain and were reduced under negative C gain. 13 C data showed that C was allocated to storage pools independent of the net flux and even under severe C limitation. Allocation to below-ground tissues was strongest in control trees followed by trees experiencing drought followed by those grown under low [CO 2 ]. Our data suggest that NSC storage has, under the conditions of our experimental manipulation (e.g., strong progressive drought, no above-ground growth), a high allocation priority and cannot be considered an overflow process. While these results also suggest active storage allocation, definitive proof of active plant control of storage in woody plants requires studies involving molecular tools.

Description: This study quantified the effect of soil warming on sap flow density ( Q s ) of Pinus cembra L. at the treeline in the Central Tyrolean Alps. To enhance soil temperature we installed a transparent roof construction above the forest floor around six trees. Six other trees served as controls in the absence of any manipulation. Roofing enhanced growing season mean soil temperature by 1.6, 1.3 and 1.0 °C at 5, 10 and 20 cm soil depth, respectively, while soil water availability was not affected. Sap flow density (using Granier-type thermal dissipation probes) and environmental parameters were monitored throughout three growing seasons. During the first year of treatment, no warming effect was detected on Q s . However, soil warming caused Q s to increase significantly by 11 and 19% above levels in control trees during the second and third year, respectively. This effect appeared to result from warming-induced root production, a reduction in viscosity and perhaps an increase also in root hydraulic conductivity. Hardly affected were leaf-level net CO 2 uptake rate and conductance for water vapour, so that water-use efficiency stayed unchanged as confirmed by needle 13 C analysis. We conclude that tree water loss will increase with soil warming, which may alter the water balance within the treeline ecotone of the Central Austrian Alps in a future warming environment.

Description: Process-based models that link seasonally varying environmental signals to morphological features within tree rings are essential tools to predict tree growth response and commercially important wood quality traits under future climate scenarios. This study evaluated model portrayal of radial growth and wood anatomy observations within a mature maritime pine ( Pinus pinaster (L.) Aït.) stand exposed to seasonal droughts. Intra-annual variations in tracheid anatomy and wood density were identified through image analysis and X-ray densitometry on stem cores covering the growth period 1999–2010. A cambial growth model was integrated with modelled plant water status and sugar availability from the soil–plant–atmosphere transfer model MuSICA to generate estimates of cell number, cell volume, cell mass and wood density on a weekly time step. The model successfully predicted inter-annual variations in cell number, ring width and maximum wood density. The model was also able to predict the occurrence of special anatomical features such as intra-annual density fluctuations (IADFs) in growth rings. Since cell wall thickness remained surprisingly constant within and between growth rings, variations in wood density were primarily the result of variations in lumen diameter, both in the model and anatomical data. In the model, changes in plant water status were identified as the main driver of the IADFs through a direct effect on cell volume. The anatomy data also revealed that a trade-off existed between hydraulic safety and hydraulic efficiency. Although a simplified description of cambial physiology is presented, this integrated modelling approach shows potential value for identifying universal patterns of tree-ring growth and anatomical features over a broad climatic gradient.

Description: Selecting plantation species to balance water use and production requires accurate models for predicting how species will tolerate and respond to environmental conditions. Although interspecific variation in water use occurs, species-specific parameters are rarely incorporated into physiologically based models because often the appropriate species parameters are lacking. To determine the physiological control over water use in Eucalyptus , five stands of Eucalyptus species growing in a common garden were measured for sap flux rates and their stomatal response to vapour pressure deficit ( D ) was assessed. Maximal canopy conductance and whole-canopy stomatal sensitivity to D and reduced water availability were lower in species originating from more arid climates of origin than those from humid climates. Species from humid climates showed a larger decline in maximal sap flux density ( J Smax ) with reduced water availability, and a lower D at which stomatal closure occurred than species from more arid climates, implying larger sensitivity to water availability and D in these species. We observed significant ( P  〈 0.05) correlations of species climate of origin with mean vessel diameter ( R 2  = 0.90), stomatal sensitivity to D ( R 2  = 0.83) and the size of the decline in J Smax to restricted water availability ( R 2  = 0.94). Thus aridity of climate of origin appears to have a selective role in constraining water-use response among the five Eucalyptus plantation species. These relationships emphasize that within this congeneric group of species, climate aridity constrains water use. These relationships have implications for species choices for tree plantation success against drought-induced losses and the ability to manage Eucalyptus plantations against projected changes in water availability and evaporation in the future.

Description: Mixtures can be more productive than monocultures and may therefore use more water, which may make them more susceptible to droughts. The species interactions that influence growth, transpiration and water-use efficiency (WUE, tree growth per unit transpiration) within a given mixture vary with intra- and inter-annual climatic variability, stand density and tree size, but these effects remain poorly quantified. These relationships were examined in mixtures and monocultures of Eucalyptus globulus Labill. and Acacia mearnsii de Wildeman. Growth and transpiration were measured between ages 14 and 15 years. All E. globulus trees in mixture that were growing faster than similar sized trees in monocultures had higher WUE, while trees with similar growth rates had similar WUE. By the age of 14 years A. mearnsii trees were beginning to senesce and there were no longer any relationships between tree size and growth or WUE. The relationship between transpiration and tree size did not differ between treatments for either species, so stand-level increases in transpiration simply reflected the larger mean tree size in mixtures. Increasing neighbourhood basal area increased the complementarity effect on E. globulus growth and transpiration. The complementarity effect also varied throughout the year, but this was not related to the climatic seasonality. This study shows that stand-level responses can be the net effect of a much wider range of individual tree-level responses, but at both levels, if growth has not increased for a given species, it appears unlikely that there will be differences in transpiration or WUE for that species. Growth data may provide a useful initial indication of whether mixtures have higher transpiration or WUE, and which species and tree sizes contribute to this effect.

Description: Tree growth is frequently linked to weather conditions prior to the growing season but our understanding of these lagged climate signatures is still poorly developed. We investigated the influence of masting behaviour on the relationship between growth and climate in European Beech ( Fagus sylvatica L.) using a rare long-term dataset of seed production and a new regional tree ring chronology. Fagus sylvatica is a masting species with synchronous variations in seed production which are strongly linked to the temperature in the previous two summers. We noted that the weather conditions associated with years of heavy seed production (mast years) were the same as commonly reported correlations between growth and climate for this species. We tested the hypothesis that a trade-off between growth and reproduction in mast years could be responsible for the observed lagged correlations between growth and previous summers' temperatures. We developed statistical models of growth based on monthly climate variables, and show that summer drought (negative correlation), temperature of the previous summer (negative) and temperature of the summer 2 years previous (positive) are significant predictors of growth. Replacing previous summers' temperature in the model with annual seed production resulted in a model with the same predictive power, explaining the same variance in growth. Masting is a common behaviour in many tree species and these findings therefore have important implications for the interpretation of general climate–growth relationships. Lagged correlations can be the result of processes occurring in the year of growth (that are determined by conditions in previous years), obviating or reducing the need for ‘carry-over’ processes such as carbohydrate depletion to be invoked to explain this climate signature in tree rings. Masting occurs in many tree species and these findings therefore have important implications for the interpretation of general climate–growth relationships.

Description: The physiological response of plants growing in their natural habitat is strongly determined by seasonal variations in environmental conditions and the interaction of abiotic and biotic stresses. Here, leaf water and nutrient contents, changes in cellular redox state and endogenous levels of stress-related phytohormones (abscisic acid (ABA), salicylic acid and jasmonates) were examined during the rainy and dry season in Vellozia gigantea , an endemic species growing at high elevations in the rupestrian fields of the Espinhaço Range in Brazil. Enhanced stomatal closure and increased ABA levels during the dry season were associated with an efficient control of leaf water content. Moreover, reductions in 12- oxo -phytodienoic acid (OPDA) levels during the dry season were observed, while levels of other jasmonates, such as jasmonic acid and jasmonoyl-isoleucine, were not affected. Changes in ABA and OPDA levels correlated with endogenous concentrations of iron and silicon, hydrogen peroxide, and vitamin E, thus indicating complex interactions between water and nutrient contents, changes in cellular redox state and endogenous hormone concentrations. Results also suggested crosstalk between activation of mechanisms for drought stress tolerance (as mediated by ABA) and biotic stress resistance (mediated by jasmonates), in which vitamin E levels may serve as a control point. It is concluded that, aside from a tight ABA-associated regulation of stomatal closure during the dry season, crosstalk between activation of abiotic and biotic defences, and nutrient accumulation in leaves may be important modulators of plant stress responses in plants growing in their natural habitat.

Description: The presence of the American root-rot disease fungus Heterobasidion irregulare Garbel. & Otrosina was detected in Italian coastal pine forests ( Pinus pinea L.) in addition to the common native species Heterobasidion annosum (Fries) Brefeld. High levels of tropospheric ozone (O 3 ) as an atmospheric pollutant are usually experienced in Mediterranean pine forests. To explore the effect of interaction between the two Heterobasidion species and ozone pollution on P. pinea , an open-top chamber (OTC) experiment was carried out. Five-year-old P. pinea seedlings were inoculated with the fungal species considered ( H. irregulare , H. annosum and mock-inoculation as control), and then exposed in charcoal-filtered open-top chambers (CF-OTC) and non-filtered ozone-enriched chambers (NF+) from July to the first week of August 2010 at the experimental facilities of Curno (North Italy). Fungal inoculation effects in an ozone-enriched environment were assessed as: (i) the length of the inoculation lesion; (ii) chlorophyll a fluorescence (ChlF) responses; and (iii) analysis of resin terpenes. Results showed no differences on lesion length between fungal and ozone treatments, whereas the short-term effects of the two stress factors on ChlF indicate an increased photosynthetic efficiency, thus suggesting the triggering of compensation/repair processes. The total amount of resin terpenes is enhanced by fungal infection of both species, but depressed by ozone to the levels observed in mock-inoculated plants. Variations in terpene profiles were also induced by stem base inoculations and ozone treatment. Ozone might negatively affect terpene defences making plants more susceptible to pathogens and insects.

Description: Initial growth of germinated seeds is an important life history stage, critical for establishment and succession in forests. Important questions remain regarding the differences among species in early growth potential arising from shade tolerance. In addition, the role of leaf habit in shaping relationships underlying shade tolerance-related differences in seedling growth remains unresolved. In this study we examined variation in morphological and physiological traits among seedlings of 10 forest tree species of the European temperate zone varying in shade tolerance and leaf habit (broadleaved winter-deciduous species vs needle-leaved conifers) during a 10-week period. Seeds were germinated and grown in a controlled environment simulating an intermediate forest understory light environment to resolve species differences in initial growth and biomass allocation. In the high-resource experimental conditions during the study, seedlings increased biomass allocation to roots at the cost of leaf biomass independent of shade tolerance and leaf habit. Strong correlations between relative growth rate (RGR), net assimilation rate (NAR), leaf area ratio (LAR), specific leaf area (SLA) and leaf mass fraction (LMF) indicate that physiology and biomass allocation were equally important determinants of RGR as plant structure and leaf morphology among these species. Our findings highlight the importance of seed mass- and seed size-related root morphology (specific root length—SRL) for shade tolerance during early ontogeny. Leaf and plant morphology (SLA, LAR) were more successful in explaining variation among species due to leaf habit than shade tolerance. In both broadleaves and conifers, shade-tolerant species had lower SRL and greater allocation of biomass to stems (stem mass fraction). Light-seeded shade-intolerant species with greater SRL had greater RGR in both leaf habit groups. However, the greatest plant mass was accumulated in the group of heavy-seeded shade-tolerant broadleaves. The results of our study suggest that the combinations of plant attributes enhancing growth under high light vary with shade tolerance, but differ between leaf habit groups.

Description: Rhizospheric nitric oxide (NO) and carbon dioxide (CO 2 ) are signalling compounds known to affect physiological processes in plants. Their joint influence on tree nitrogen (N) nutrition, however, is still unknown. Therefore, this study investigated, for the first time, the combined effect of rhizospheric NO and CO 2 levels on N uptake and N pools in European beech ( Fagus sylvatica L.) seedlings depending on N availability. For this purpose, roots of seedlings were exposed to one of the nine combinations (i.e., low, ambient, high NO plus CO 2 concentration) at either low or high N availability. Our results indicate a significant effect of rhizospheric NO and/or CO 2 concentration on organic and inorganic N uptake. However, this effect depends strongly on NO and CO 2 concentration, N availability and N source. Similarly, allocation of N to different N pools in the fine roots of beech seedlings also shifted with varying rhizospheric gas concentrations and N availability.

Description: Respiration from vegetation is a substantial part of the global carbon cycle and the responses of plant respiration to daily and seasonal fluctuations in temperature and light must be incorporated in models of terrestrial respiration to accurately predict these CO 2 fluxes. We investigated how leaf respiration ( R ) responded to changes in leaf temperature ( T leaf ) and irradiance in field-grown saplings of an evergreen tree ( Eucalyptus pauciflora Sieb. ex Spreng). Seasonal shifts in the thermal sensitivity of leaf R in the dark ( R dark ) and in the light ( R light ) were assessed by allowing T leaf to vary over the day in field-grown leaves over a year. The Q 10 of R (i.e., the relative increase in R for a 10 °C increase in T leaf ) was similar for R light and R dark and had a value of ~2.5; there was little seasonal change in the Q 10 of either R light or R dark , indicating that we may be able to use similar functions to model short-term temperature responses of R in the dark and in the light. Overall, rates of R light were lower than those of R dark , and the ratio of R light / R dark tended to increase with rising T leaf , such that light suppression of R was reduced at high T leaf values, in contrast to earlier work with this species. Our results suggest we cannot assume that R light / R dark decreases with increasing T leaf on daily timescales, and highlights the need for a better mechanistic understanding of what regulates light suppression of R in leaves.

Description: Climate-related variations in functional traits of boreal tree species can result both from physiological acclimation and genetic adaptation of local populations to their biophysical environment. To improve our understanding and prediction of the physiological and growth responses of populations to climate change, we studied the role of climate of seed origin in determining variations in functional traits and its implications for tree improvement programs for a commonly reforested boreal conifer, white spruce ( Picea glauca (Moench) Voss). We evaluated growth, root-to-shoot ratio (R/S), specific leaf area (SLA), needle nitrogen ( N mass ), total non-structural carbohydrates (NSC) and photosynthetic traits of 3-year-old seedlings in a greenhouse experiment using seed from six seed orchards (SO) representing the different regions where white spruce is reforested in Québec. Height and total dry mass (TDM) were positively correlated with photosynthetic capacity ( A max ), stomatal conductance ( g s ) and mesophyll conductance ( g m ). Total dry mass, but not height growth, was strongly correlated with latitude of seed origin (SO) and associated climate variables. A max , g s , g m and more marginally, photosynthetic nitrogen-use efficiency (PNUE) were positively associated with the mean July temperature of the SO, while water use efficiency (WUE) was negatively associated. Maximum rates of carboxylation ( V cmax ), maximum rates of electron transport ( J max ), SLA, N mass , NSC and R/S showed no pattern. Our results did not demonstrate a higher A max for northern seed orchards, although this has been previously hypothesized as an adaptation mechanism for maintaining carbon uptake in northern regions . We suggest that g s , g m , WUE and PNUE are the functional traits most associated with fine-scale geographic clines and with the degree of local adaptation of white spruce populations to their biophysical environments. These geographic patterns may reflect in situ adaptive genetic differences in photosynthetic efficiency along the cline.

Description: We evaluated the long-term (1995–2008) trends in foliar and sapwood metabolism, soil solution chemistry and tree mortality rates in response to chronic nitrogen (N) additions to pine and hardwood stands at the Harvard Forest Long Term Ecological Research (LTER) site. Common stress-related metabolites like polyamines (PAs), free amino acids (AAs) and inorganic elements were analyzed for control, low N (LN, 50 kg NH 4 NO 3  ha –1  year –1 ) and high N (HN, 150 kg NH 4 NO 3  ha –1  year –1 ) treatments. In the pine stands, partitioning of excess N into foliar PAs and AAs increased with both N treatments until 2002. By 2005, several of these effects on N metabolites disappeared for HN, and by 2008 they were mostly observed for LN plot. A significant decline in foliar Ca and P was observed mostly with HN for a few years until 2005. However, sapwood data actually showed an increase in Ca, Mg and Mn and no change in PAs in the HN plot for 2008, while AAs data revealed trends that were generally similar to foliage for 2008. Concomitant with these changes, mortality data revealed a large number of dead trees in HN pine plots by 2002; the mortality rate started to decline by 2005. Oak trees in the hardwood plot did not exhibit any major changes in PAs, AAs, nutrients and mortality rate with LN treatment, indicating that oak trees were able to tolerate the yearly doses of 50 kg NH 4 NO 3 ha –1  year –1 . However, HN trees suffered from physiological and nutritional stress along with increased mortality in 2008. In this case also, foliar data were supported by the sapwood data. Overall, both low and high N applications resulted in greater physiological stress to the pine trees than the oaks. In general, the time course of changes in metabolic data are in agreement with the published reports on changes in soil chemistry and microbial community structure, rates of soil carbon sequestration and production of woody biomass for this chronic N study. This correspondence of selected metabolites with other measures of forest functions suggests that the metabolite analyses are useful for long-term monitoring of the health of forest trees.

Description: To buffer against the high spatial and temporal heterogeneity of the riparian habitat, riparian tree species, such as black poplar ( Populus nigra L.), may display a high level of genetic variation and phenotypic plasticity for functional traits. Using a multisite common garden experiment, we estimated the relative contribution of genetic and environmental effects on the phenotypic variation expressed for individual leaf area, leaf shape, leaf structure and leaf carbon isotope discrimination ( 13 C) in natural populations of black poplar. Twenty-four to 62 genotypes were sampled in nine metapopulations covering a latitudinal range from 48°N to 42°N in France and in Italy and grown in two common gardens at Orléans (ORL) and at Savigliano (SAV). In the two common gardens, substantial genetic variation was expressed for leaf traits within all metapopulations, but its expression was modulated by the environment, as attested by the genotype x environment ( G x E ) interaction variance being comparable to or even greater than genetic effects. For LA, G x E interactions were explained by both changes in genotype ranking between common gardens and increased variation in SAV, while these interactions were mainly attributed to changes in genotype ranking for 13 C. The nine P. nigra metapopulations were highly differentiated for LA, as attested by the high coefficient of genetic differentiation ( Q ST = 0.50 at ORL and 0.51 at SAV), and the pattern of metapopulation differentiation was highly conserved between the two common gardens. In contrast, they were moderately differentiated for 13 C ( Q ST = 0.24 at ORL and 0.25 at SAV) and the metapopulation clustering changed significantly between common gardens. Our results evidenced that the nine P. nigra metapopulations present substantial genetic variation and phenotypic plasticity for leaf traits, which both represent potentially significant determinants of populations' capacities to respond, on a short-term basis and over generations, to environmental variations.

Description: Conifers have incurred high mortality during recent global-change-type drought(s) in the western USA. Mechanisms of drought-related tree mortality need to be resolved to support predictions of the impacts of future increases in aridity on vegetation. Hydraulic failure, carbon starvation and lethal biotic agents are three potentially interrelated mechanisms of tree mortality during drought. Our study compared a suite of measurements related to these mechanisms between 49 mature piñon pine ( Pinus edulis Engelm.) trees that survived severe drought in 2002 (live trees) and 49 trees that died during the drought (dead trees) over three sites in Arizona and New Mexico. Results were consistent over all sites indicating common mortality mechanisms over a wide region rather than site-specific mechanisms. We found evidence for an interactive role of hydraulic failure, carbon starvation and biotic agents in tree death. For the decade prior to the mortality event, dead trees had twofold greater sapwood cavitation based on frequency of aspirated tracheid pits observed with scanning electron microscopy (SEM), smaller inter-tracheid pit diameter measured by SEM, greater diffusional constraints to photosynthesis based on higher wood 13 C, smaller xylem resin ducts, lower radial growth and more bark beetle (Coleoptera: Curculionidae) attacks than live trees. Results suggest that sapwood cavitation, low carbon assimilation and low resin defense predispose piñon pine trees to bark beetle attacks and mortality during severe drought. Our novel approach is an important step forward to yield new insights into how trees die via retrospective analysis.

Description: Knowledge of copepod in situ diet is critical for accurate assessment of trophic linkages and transfer efficiencies of the marine food web but is limited due to technical challenges. Here we report, using a recently developed eukaryote-universal copepod-excluding ectobiotic ciliate-blocking protocol, to investigate the natural diets of the copepods Temora turbinata , Subeucalanus subcrassus and Canthocalanus pauper in coastal waters in Sanya Bay, China. Analysis of the resultant 18S rDNA clone libraries revealed diverse diet composition for all the three copepod species, with 11 prey species for C. pauper , 9 for T. turbinata and 9 for S. subcrassus . The ingested materials included land plants, green algae, Metazoa, Euglenozoa and Rhizaria, although species numbers from each of these lineages differed. Broussonetia sp. (land plant), which might have been ingested in the form of pollen or fresh detritus were common among all three copepods, and accounted for a significant proportion (〉55%) of the clones sequenced. These results suggest that copepods in Sanya coastal waters might use terrigenous detritus as supplementary food sources when phytoplankton production is limited. However, the significance of the plant detritus as a nutrition source of these copepods remains to be determined by analyzing carbon-based proportion and digestion/assimilation rate of the ingested plant materials.

Description: Predation is considered an important source of mortality for plankton, but documenting variation in planktonic predation, particularly across interacting environmental cycles, remains logistically difficult, thus our understanding remains limited. To test for the combined effects of prey life history stage, diel or light level phase (including crepuscular periods) and seasonal upwelling on the risk of predation, we deployed tethered adult and larval brine shrimp Artemia franciscana using dock-based plankton tethering units (PTUs). Risk was higher overall during upwelling, but life history stage also interacted with season. There was no seasonal difference in risk for adults. Larvae were at significantly lower risk of predation during non-upwelling than during upwelling. Larvae were also at lower risk during non-upwelling than were adults during either season. During upwelling, there was no significant difference in risk between the two prey categories. With respect to the diel cycle, dusk was safer than daytime. For larvae, the diel pattern in risk remained consistent across seasons while risk for adults at night was slightly lower during upwelling than during non-upwelling. Variation in planktonic predation risk across diel and seasonal cycles differs for different life history stages and thus, generalizations fail to capture the complexity of interactions between factors.

Description: Protistan grazers and viruses are major agents of mortality in marine microbial communities with substantially different implications for food-web dynamics, carbon cycling and diversity maintenance. While grazers and viruses are typically studied independently, their impacts on microbial communities may be complicated by direct and indirect interactions of their mortality effects. Using a modification of the seawater dilution approach, we quantified growth and mortality rates for total phytoplankton and picophytoplankton populations ( Prochlorococcus , Synechococcus , picoeukaryotes) at four contrasting sites in the California Current Ecosystem. Grazing mortality was significant in 10 of 15 cases, while viral effects were significant for 2 cases. Nonetheless, mortality estimates for the entire phytoplankton community based on chlorophyll a were 38 ± 13% higher when viral effects were included, relative to grazing alone. Mortality estimates for picophytoplankton varied in space and among groups. We also explored a potential methodological constraint of this method and hypothesize that heterotrophic bacteria may be affected by the dilution of their growth-sustaining substrates. For all picophytoplankton, estimates of grazing and viral mortality were inversely related within and across experiments. Indirect interactions among grazers and viruses may be important to consider if there are tradeoffs in the grazing and virus resistance strategies of prey/host cells.

Description: Dispersal of native species from the regional pool can recover invaded communities to a pre-invaded state by supplementing declining populations or providing resistant species. However, dispersal may also exacerbate the negative effects of an invader. Introduced species can open or create new niche space, which could facilitate the establishment of competitors or predators that previously could not succeed in the uninvaded local community. To investigate the interaction between dispersal and invasion by a non-native consumer, we conducted a field mesocosm experiment that introduced zebra mussels into native zooplankton communities. Regional zooplankton were collected and added to both invaded and uninvaded communities. In uninvaded communities, zooplankton dispersal reduced cladoceran diversity by ~40%, rotifer abundance by ~65% and copepod nauplii abundance by ~80%. In invaded communities, dispersal increased cladoceran diversity by ~60%, but also further exacerbated the negative effects of zebra mussels on rotifer abundance. This experiment illustrates the potential for dispersal to both positively and negatively affect local communities, and how these effects may change with disturbance and the taxa or community metric of study.

Description: Pulse amplitude-modulated (PAM) fluorometry was used to obtain rapid light curves (RLCs) for phytoplankton from small Canadian lakes of varying water clarity, including metalimnetic communities from two clear lakes. RLCs were measured before and after exposure to a number of experimental spectra containing ultraviolet radiation (UVR) and photosynthetically active radiation (PAR). Photochemical (qP) and non-photochemical (NPQ) quenching during RLCs were positively and significantly correlated with recent in situ light history, but the maximum quantum efficiency of Photosystem II (α) and maximum relative rate of electron transport (rETR max ) were not. rETR max and α were diminished by experimental exposures to UVR and/or high PAR, but significantly less so in phytoplankton from brighter environments. UVR exposures diminished the inducible (photoprotective) NPQ of most epilimnetic phytoplankton. Both the inducible and total (photoprotective + photoinhibitory) NPQ of metalimnetic phytoplankton were stimulated by spectral exposures. Our results are the first obtained from natural communities of freshwater phytoplankton to show that aspects of PSII photophysiology (as inferred from RLCs) vary according to in situ light history.

Description: Inducible defenses are a common phenotypically plastic response to a heterogeneous predation risk. Once induced, these defenses cannot only lose their benefit, but even become costly, should the predator disappear. Consequently, some organisms have developed the ability to reverse their defensive traits. However, despite extensive research on inducible defenses, reports on reversibility are rare and mostly concentrate on defensive behavior. In our study, we investigated the reversibility of morphological defenses in the freshwater crustacean Daphnia barbata . This species responds to Notonecta glauca and Triops cancriformis with two distinctively defended morphotypes. Within the numerous defensive traits, we found both trait- and predator-specific reversibility. Body torsion and tail-spine-related traits were highly reversible, whereas helmet-related traits remained stable, suggesting different physiological constraints. However, in general, we found the defenses against Triops mostly reversible, while Notonecta -induced defenses were persistent and grew further, even in the absence of a predator.

Description: Marine phytoplankton are a taxonomically and functionally diverse group of organisms that are key players in the most important biogeochemical cycles. Phytoplankton taxa show different resource utilization strategies (e.g. nutrient-uptake rates and cellular allocation) and traits. Therefore, acknowledging this diversity is crucial to understanding how elemental cycles operate, including the origin and dynamics of elemental ratios. In this paper, we focus on trait-based models as tools to study the role of phytoplankton diversity in the stoichiometric phenomenology observed in the laboratory and in the open ocean. We offer a compilation of known empirical results on stoichiometry and summarize how trait-based approaches have attempted to replicate these results. By contrasting the different ecological and evolutionary approaches available in the literature, we explore the strengths and limitations of the existing models. We thus try to identify existing gaps and challenges, and point to potential new directions that can be explored to fill these gaps. We aim to highlight the potential of including diversity explicitly in our modeling approaches, which can help us gain important knowledge about changes in local and global stoichiometric patterns.

Description: Highly intermittent spatial variability of phytoplankton is observed ubiquitously in marine ecosystems, especially when measurements are performed at the micro-scale level. Therefore, theoretical developments and new modelling tools are required to understand the observed small-scale vertical structure and its relationship to ecosystem behaviour. Nearly all current ecosystem models are formulated entirely based on the mean field approximation, ignoring sub-grid scale variability. Even if such approximation may be reasonable for meso-scales (and above), it cannot account for micro-scale dynamics, which may also impact macroscopic properties at the larger scale. To consider intermittency of variables in plankton ecosystem models, we apply a newly developed modelling approach called the closure approach. Detailed simulations were conducted, combining fluid-dynamics of the 1D water column with the nutrient-phytoplankton closure ecosystem model for application to a site in the northern North Sea. Compared with a control model, which does not account for such intermittency, the closure model produced substantially different spatio-temporal patterns of mean phytoplankton biomass and growth rate, which depended on the overall level of variability. In this study, we (i) seek to explore the effects of sub-scale variability coupled with physical transport and (ii) begin to address the yet unresolved question of how to consistently model the advection and diffusion of the variances and co-variances used to represent sub-scale variability in the closure approach. Our results suggest that it may be necessary to account explicitly for the intermittent distribution of plankton and nutrients, even in large-scale biogeochemical models.

Description: Seawater viscosity is influenced by temperature as well as through excretion of exopolymers by some plankton. We examined the role of viscosity on the movement patterns and sensory abilities of the dinoflagellate Heterocapsa triquetra , manipulating the viscosity of seawater to simulate a 10 ± 1.5°C temperature change. In a second treatment, we seeded the water with microbeads to examine swimming behaviours in the presence of a mechanical stimulus. Increased viscosity reduced distances between conspecifics 4.7-fold and increased distances between protists and microbeads by 3.4-fold. Increased viscosity also affected other aspects of motility, with an overall reduction in swimming speed of 2.0- and 7.0-fold for treatments with and without mechanical stimuli. Higher viscosities were associated with upward vertical migration, in both the presence and absence of microbeads. Cells were highly sensitive to disturbances to the velocity field, by as little as 1.5%, and different approach distances of H. triquetra to conspecifics over mechanical stimuli suggest sensory capacity to distinguish types of particles. Mediation of motility and migratory behaviours through viscosity implies ramifications for the distribution of protists and their encounters with resources, predators and conspecifics triggered by events such as temperature changes and phytoplankton bloom events.

Description: High-resolution stem diameter variations (SDV) are widely recognized as a useful drought stress indicator and have therefore been used in many irrigation scheduling studies. More recently, SDV have been used in combination with other plant measurements and biophysical modelling to study fundamental mechanisms underlying whole-plant functioning and growth. The present review aims to scrutinize the important insights emerging from these more recent SDV applications to identify trends in ongoing fundamental research. The main mechanism underlying SDV is variation in water content in stem tissues, originating from reversible shrinkage and swelling of dead and living tissues, and irreversible growth. The contribution of different stem tissues to the overall SDV signal is currently under debate and shows variation with species and plant age, but can be investigated by combining SDV with state-of-the-art technology like magnetic resonance imaging. Various physiological mechanisms, such as water and carbon transport, and mechanical properties influence the SDV pattern, making it an extensive source of information on dynamic plant behaviour. To unravel these dynamics and to extract information on plant physiology or plant biophysics from SDV, mechanistic modelling has proved to be valuable. Biophysical models integrate different mechanisms underlying SDV, and help us to explain the resulting SDV signal. Using an elementary modelling approach, we demonstrate the application of SDV as a tool to examine plant water relations, plant hydraulics, plant carbon relations, plant nutrition, freezing effects, plant phenology and dendroclimatology. In the ever-expanding SDV knowledge base we identified two principal research tracks. First, in detailed short-term experiments, SDV measurements are combined with other plant measurements and modelling to discover patterns in phloem turgor, phloem osmotic concentrations, root pressure and plant endogenous control. Second, long-term SDV time series covering many different species, regions and climates provide an expanding amount of phenotypic data of growth, phenology and survival in relation to microclimate, soil water availability, species or genotype, which can be coupled with genetic information to support ecological and breeding research under on-going global change. This under-exploited source of information has now encouraged research groups to set up coordinated initiatives to explore this data pool via global analysis techniques and data-mining.

Description: Even-aged forest stands are competitive communities where competition for light gives advantages to tall individuals, thereby inducing a race for height. These same individuals must however balance this competitive advantage with height-related mechanical and hydraulic risks. These phenomena may induce variations in height–diameter growth relationships, with primary dependences on stand density and tree social status as proxies for competition pressure and access to light, and on availability of local environmental resources, including water. We aimed to investigate the effects of stand density, tree social status and water stress on the individual height–circumference growth allocation ( h – c ), in even-aged stands of Quercus petraea Liebl. (sessile oak). Within-stand c was used as surrogate for tree social status. We used an original long-term experimental plot network, set up in the species production area in France, and designed to explore stand dynamics on a maximum density gradient. Growth allocation was modelled statistically by relating the shape of the h – c relationship to stand density, stand age and water deficit. The shape of the h – c relationship shifted from linear with a moderate slope in open-grown stands to concave saturating with an initial steep slope in closed stands. Maximum height growth was found to follow a typical mono-modal response to stand age. In open-grown stands, increasing summer soil water deficit was found to decrease height growth relative to radial growth, suggesting hydraulic constraints on height growth. A similar pattern was found in closed stands, the magnitude of the effect however lowering from suppressed to dominant trees. We highlight the high phenotypic plasticity of growth in sessile oak trees that further adapt their allocation scheme to their environment. Stand density and tree social status were major drivers of growth allocation variations, while water stress had a detrimental effect on height in the h – c allocation.

Description: Phenological synchronisms between apical and lateral meristems could clarify some aspects related to the physiological relationships among the different organs of trees. This study correlated the phenological phases of bud development and xylem differentiation during spring 2010–14 in balsam fir ( Abies balsamea Mill.) and black spruce [( Picea mariana Mill. (BSP)] of the Monts-Valin National Park (Quebec, Canada) by testing the hypothesis that bud development occurs after the reactivation of xylem growth. From May to September, we conducted weekly monitoring of xylem differentiation using microcores and bud development with direct observations on terminal branches. Synchronism between the beginning of bud development and xylem differentiation was found in both species with significant correlations between the phases of bud and xylem phenology. Degree-day sum was more appropriate in assessing the date of bud growth resumption, while thermal thresholds were more suitable for cambium phenology. Our results provide new knowledge on the dynamics of spring phenology and novel information on the synchronisms between two meristems in coniferous trees. The study demonstrates the importance of precisely defining the phases of bud development in order to correctly analyse the relationships with xylem phenology.

Description: Salicylic acid (SA) is a defense-related key signaling molecule involved in plant immunity. In this study, a subgroup IIa WRKY gene PtrWRKY40 was isolated from Populus trichocarpa , which displayed amino acid sequence similar to Arabidopsis AtWRKY40 , AtWRKY18 and AtWRKY60. PtrWRKY40 transcripts accumulated significantly in response to SA, methyl jasmonate and hemibiotrophic fungus Dothiorella gregaria Sacc. Overexpression of PtrWRKY40 in transgenic poplar conferred higher susceptibility to D. gregaria infection. This susceptibility was coupled with reduced expression of SA-associated genes ( PR1.1 , PR2.1 , PR5.9 , CPR5 and SID2 ) and jasmonic acid (JA)-related gene JAZ8 . Decreased accumulation of endogenous SA was observed in transgenic lines overexpressing PtrWRKY40 when compared with wild-type plants. However, constitutive expression of PtrWRKY40 in Arabidopsis thaliana displayed resistance to necrotrophic fungus Botrytis cinerea , and the expression of JA-defense-related genes such as PDF1.2 , VSP2 and PR3 was remarkably increased in transgenic plants upon infection with fugal pathogens. Together, our findings indicate that PtrWRKY40 plays a negative role in resistance to hemibiotrophic fungi in poplar but functions as a positive regulator of resistance toward the necrotrophic fungi in Arabidopsis .

Description: Urea fertilization decreases Pinus taeda L. growth in clay soils of subtropical areas. The negative effect of urea is related to changes in some hydraulic traits, similar to those observed in plants growing under drought. The aims of this work were (i) to determine whether different sources of nitrogen applied as fertilizers produce similar changes in growth and hydraulic traits to those observed by urea fertilization and (ii) to analyze the impact of those changes in plant drought tolerance. Plants fertilized with urea, nitrate (NO3–) or ammonium (NH4+) were grown well watered or with reduced water supply. Urea and NO3– fertilization reduced plant growth and increased root hydraulic conductance scaled by root dry weight (DW). NH4+ fertilization did not reduce plant growth and increased shoot hydraulic conductance and stem hydraulic conductivity. We conclude that NO3– is the ion involved in the changes linked to the negative effect of urea fertilization on P. taeda growth. NH4+ fertilization does not change drought susceptibility and it produces changes in shoot hydraulic traits, therefore plants avoid the depressive effect of fertilization. Urea and NO3– fertilizers induce changes in DW and root hydraulic conductance and consequently plants are less affected by drought.

Description: It is unclear how or even if phosphorus (P) input alters the influence of nitrogen (N) deposition in a forest. In theory, nutrients in leaves and twigs differing in age may show different responses to elevated nutrient input. To test this possibility, we selected Chinese fir ( Cunninghamia lanceolata ) for a series of N and P addition experiments using treatments of +N1 – P (50 kg N ha –1 year –1 ), +N2 – P (100 kg N ha –1 year –1 ), –N + P (50 kg P ha –1 year –1 ), +N1 + P, +N2 + P and –N – P (without N and P addition). Soil samples were analyzed for mineral N and available P concentrations. Leaves and twigs in summer and their litters in winter were classified as and sorted into young and old components to measure N and P concentrations. Soil mineral N and available P increased with N and P additions, respectively. Nitrogen addition increased leaf and twig N concentrations in the second year, but not in the first year; P addition increased leaf and twig P concentrations in both years and enhanced young but not old leaf and twig N accumulations. Nitrogen and P resorption proficiencies in litters increased in response to N and P additions, but N and P resorption efficiencies were not significantly altered. Nitrogen resorption efficiency was generally higher in leaves than in twigs and in young vs old leaves and twigs. Phosphorus resorption efficiency showed a minimal variation from 26.6 to 47.0%. Therefore, P input intensified leaf and twig N enrichment with N addition, leaf and twig nutrients were both gradually resorbed with aging, and organ and age effects depended on the extent of nutrient limitation.

Description: Manipulating tree belowground carbon (C) transport enables investigation of the ecological and physiological roles of tree roots and their associated mycorrhizal fungi, as well as a range of other soil organisms and processes. Girdling remains the most reliable method for manipulating this flux and it has been used in numerous studies. However, girdling is destructive and irreversible. Belowground C transport is mediated by phloem tissue, pressurized through the high osmotic potential resulting from its high content of soluble sugars. We speculated that phloem transport may be reversibly blocked through the application of an external pressure on tree stems. Thus, we here introduce a technique based on compression of the phloem, which interrupts belowground flow of assimilates, but allows trees to recover when the external pressure is removed. Metal clamps were wrapped around the stems and tightened to achieve a pressure theoretically sufficient to collapse the phloem tissue, thereby aiming to block transport. The compression's performance was tested in two field experiments: a 13 C canopy labelling study conducted on small Scots pine ( Pinus sylvestris L.) trees [2–3 m tall, 3–7 cm diameter at breast height (DBH)] and a larger study involving mature pines (~15 m tall, 15–25 cm DBH) where stem respiration, phloem and root carbohydrate contents, and soil CO 2 efflux were measured. The compression's effectiveness was demonstrated by the successful blockage of 13 C transport. Stem compression doubled stem respiration above treatment, reduced soil CO 2 efflux by 34% and reduced phloem sucrose content by 50% compared with control trees. Stem respiration and soil CO 2 efflux returned to normal within 3 weeks after pressure release, and 13 C labelling revealed recovery of phloem function the following year. Thus, we show that belowground phloem C transport can be reduced by compression, and we also demonstrate that trees recover after treatment, resuming C transport in the phloem.

Description: The Mediterranean region is an area of special interest for conservation where the incidence of multiple drivers of global change is expected to increase. One of the factors predicted to change is soil-nutrient availability, an essential factor for plant growth. Thus, study of the effects of variation in this parameter is especially relevant in species with a circum-Mediterranean distribution, such as Arbutus unedo L., in which the different provenances grow in different habitats, which must differ in nutritional conditions. We aimed to determine the effect of provenance on plasticity, to establish whether structural and morphological traits differ in the level of plasticity and to assess how nutrients affect the photosynthetic light response. In a common garden experiment, we studied seven provenances from the circum-Mediterranean range of A. unedo and established two nutrient treatments (low and high nutrient availability). We measured physiological and structural traits in 1-year-old sapling and determined a phenotypic plasticity index (PPI) to quantify the level of plasticity, whereas the radiation effects were tested by construction and analysis of light response curves. Interestingly, provenance did not explain a significant amount of variance, but the plasticity was four times higher for the structural traits than for the physiological traits. Therefore, the plasticity to nutrient availability will not favour or prevent the expansion or contraction of the range of any of these provenances of A. unedo . Furthermore, the structural plasticity demonstrated the ability of the strawberry tree to optimize resource allocation, whereas the physiology remained stable, thus avoiding extra expenditure. The study findings also suggest that increased availability of nutrients would improve the performance of the species during the Mediterranean summer, characterized by high irradiance. These abilities will be key to the survival of saplings of the species under the future scenario of changes in nutrient availability.

Description: An increase in temperature along with a decrease in summer precipitation in Central Europe will result in an increased frequency of drought events and gradually lead to a change in species composition in forest ecosystems. In the present study, young oaks ( Quercus robur L. and Quercus petraea (Matt.) Liebl.) were transplanted into large mesocosms and exposed for 3 years to experimental warming and a drought treatment with yearly increasing intensities. Carbon and oxygen isotopic ( 13 C and 18 O) patterns were analysed in leaf tissue and tree-ring cellulose and linked to leaf physiological measures and tree-ring growth. Warming had no effect on the isotopic patterns in leaves and tree rings, while drought increased 18 O and 13 C. Under severe drought, an unexpected isotopic pattern, with a decrease in 18 O, was observed in tree rings but not in leaves. This decrease in 18 O could not be explained by concurrent physiological analyses and is not supported by current physiological knowledge. Analysis of intra-annual tree-ring growth revealed a drought-induced growth cessation that interfered with the record of isotopic signals imprinted on recently formed leaf carbohydrates. This missing record indicates isotopic uncoupling of leaves and tree rings, which may have serious implications for the interpretation of tree-ring isotopes, particularly from trees that experienced growth-limiting stresses.