Leaf and tree age-related changes in leaf ecophysiological traits, nutrient, and adaptive strategies of Alnus nepalensis in the central Himalaya.

Leaf ecophysiological traits are known to change with leaf and tree age. In the present study, we measured the effect of leaf and tree age on leaf ecophysiological and morphological traits of nitrogen-fixing Alnus nepalensis (D. Don) which is a pioneer tree species in degraded lands. Three naturally occurring A. nepalensis forest stands, namely young (5-8 years old), mature (40-55 years old), and old (130-145 years old), were considered in this study. We also investigated the seasonal variations in leaf ecophysiological and morphological traits during leaf flushing, fully expanded, and leaf senescence phenological stages. The ecophysiological and morphological traits were compared between leaf and tree ages using a linear mixed-effect model (LMM) and Tukey's HSD test. Fully expanded leaves and young trees demonstrate ecophysiological traits consistent with acquisitive resource-use strategies. Our results revealed that net photosynthetic capacity (Aarea and Amass), leaf stomatal conductance (gswarea and gswmass), transpiration rate (Earea and Emass), specific leaf area (SLA), predawn and midday water potential (Ψ), leaf total chlorophyll concentration, photosynthetic N- and P-use efficiency (PNUE and PPUE) were higher in younger trees than mature and old trees. We found lower wateruse efficiency (WUE) and intrinsic water-use efficiency (WUEi) in young trees than in mature and old ones. Mass-based net photosynthetic capacity (Amass) was positively correlated with PNUE, PPUE, transpiration rate, stomatal conductance, SLA and chlorophyll concentrations but negatively correlated with WUE and WUEi. However, mass-based leaf nitrogen (N) and phosphorus (P) concentrations were the highest in fully expanded leaves and did not vary with tree age despite N concentration being negatively correlated with SLA. Overall, this study provides valuable insights into the age-related changes in leaf ecophysiological traits of A. nepalensis. The findings underscore the importance of considering tree age when studying plant ecophysiology and highlight the acquisitive resource-use strategies employed by young trees for rapid growth and establishment.

Ecosystem carbon stocks and sequestration rates in white oak forests in the central Himalaya: Role of nitrogen-fixing Nepalese alder.

Nitrogen-fixing Nepalese alder (Alnus nepalensis D. Don.), a pioneer species and nurse tree species, forms pure stands, and sometimes occurs in mixed stands in areas affected by landslides. The objective of this study was to understand the influence of A. nepalensis on carbon stock in white oak (Quercus leucotrichophora A. Camus) forests. We investigated the differences in vegetation biomass carbon (tree, sapling, seedling, shrub and herbs, and forest floor mass), soil organic carbon stock, and sequestration rates in five naturally occurring oak mixed alder (OMA) forest stands and five naturally occurring oak without alder (OWA) forest stands along the basal area gradient in order to investigate the role of A. nepalensis on ecosystem carbon stock. The total basal area ranged from 61.20 to 89.51 m2 ha-1 in the OMA stands and from 38.02 to 53.54 m2 ha-1 in the OWA stands. The total tree density of the OMA stands (1120 to 1330 trees ha-1) was higher than that of the OWA stands (950 to 1230 trees ha-1). The total ecosystem carbon stock in the OMA stands was significantly (P<0.05) higher than that in the OWA stands, ranging from 485.3 to 635.6 Mg C ha-1 in the former and from 378.8 to 472 Mg C ha-1 in the latter. Soil was the second largest carbon pool in all the studied stands, with the values ranging from 238.1 to 254.1 Mg C ha-1 in the OMA and 185.5 to 215.8 Mg C ha-1 in the OWA stands. The soil organic carbon (SOC) stock was 1.19 to 1.28 times higher in the OMA than in the OWA stands. Of the total ecosystem carbon stock in different OMA stands, A. nepalensis stored 16.2 to 38.8%. Annual carbon sequestration rates (6.6 to 9.5 Mg C ha-1 yr-1) in the OMA stands were significantly (P<0.05) higher than in the OWA (2.5 to 5.4 Mg C ha-1 yr-1) stands. Among all the species and across the stands, the greatest carbon sequestration was exhibited by A. nepalensis (3.4 to 5.3 Mg C ha-1 yr-1). The present results show the role of A. nepalensis in ecosystem carbon stock and sequestration rates. Significantly higher rates of carbon sequestration by oak in OMA stands than OWA stands clearly indicate the facilitative role of co-occurring nitrogen-fixing A. nepalensis. The results imply that Q. leucotrichophora mixed with a A. nepalensis plantation may be a good option for enhancing ecosystem carbon stock, carbon sequestration, and habitat restoration in the central Himalaya.

Annotated genome sequence of a fast-growing diploid clone of red alder (Alnus rubra Bong.).

Red alder (Alnus rubra Bong.) is an ecologically significant and important fast-growing commercial tree species native to western coastal and riparian regions of North America, having highly desirable wood, pigment, and medicinal properties. We have sequenced the genome of a rapidly growing clone. The assembly is nearly complete, containing the full complement of expected genes. This supports our objectives of identifying and studying genes and pathways involved in nitrogen-fixing symbiosis and those related to secondary metabolites that underlie red alder's many interesting defense, pigmentation, and wood quality traits. We established that this clone is most likely diploid and identified a set of SNPs that will have utility in future breeding and selection endeavors, as well as in ongoing population studies. We have added a well-characterized genome to others from the order Fagales. In particular, it improves significantly upon the only other published alder genome sequence, that of Alnus glutinosa. Our work initiated a detailed comparative analysis of members of the order Fagales and established some similarities with previous reports in this clade, suggesting a biased retention of certain gene functions in the vestiges of an ancient genome duplication when compared with more recent tandem duplications.

A Lignan from Alnus japonica Activates Myogenesis and Alleviates Dexamethasone-induced Myotube Atrophy.

To find inhibitors against skeletal muscle loss, we isolated a lignan compound ((-)-(2R,3R-1,4-O-diferuloylsecoisolarciresinol, DFS) from the stem of Alnus japonica. C2C12 myoblasts were treated with DFS during differentiation. To induce an in vitro atrophic condition, differentiated myotubes were treated with dexamethasone (a synthetic glucocorticoid). DFS (10 nM) increased expression levels of myogenic factors and the number of multi-nucleated myotubes expressing myosin heavy chain (MHC). The myogenic potential of DFS could be attributed to p38 MAPK activation. DFS also protected against dexamethasone-induced damage, showing increased expression of MHC and mammalian target of rapamycin (mTOR), a major anabolic factor. Under atrophic condition, the anti-myopathy effect of DFS was associated with inactivation of NF-κB signaling pathway and the subsequent suppression of muscle degradative E3 ligases and myostatin. DFS treatment also restored fast muscle fiber (type II a, II b, and II x), known to be susceptible to dexamethasone. These results indicate that DFS isolated from A. japonica can stimulate myogenesis via p38 MAPK activation and alleviate muscle atrophy by modulating the expression of genes associated with muscle protein anabolism/catabolism. Thus, we propose that DFS can be used as a pharmacological and nutraceutical agent for increasing muscle strength or protecting muscle loss.

Anti-Herbivore Activity of Oregonin, a Diarylheptanoid Found in Leaves and Bark of Red Alder (Alnus rubra).

Plants synthesize a wide range of bioactive secondary metabolites to defend against pests and pathogens. Red alder (Alnus rubra) bark, root, and leaf extract have a long history of use in traditional medicine and hygiene. Diarylheptanoids, especially oregonin ((5S)-1,7-bis(3,4-dihydroxyphenyl)-5-(ß-D-xylopyranosyloxy)-heptan-3-one), have been identified as major bioactive constituents. Diarylheptanoids have become a focus of research following reports of their antioxidant, antifungal, and anti-cancer activities. Recent data suggest that high oregonin concentration is associated with resistance of red alder leaves to western tent caterpillar (Malacosoma californicum) defoliation. Here we test effects of this compound directly on leaf-eating insects. Purified oregonin was examined in insect choice and toxicity tests using lepidopteran caterpillars. The compound exhibited significant anti-feedant activity against cabbage looper (Trichoplusia ni), white-marked tussock moth (Orgyia leucostigma), fall webworm (Hyphantria cunea), and M. californicum at concentrations corresponding to oregonin content of the most resistant alder clones in previous experiments. Toxicity tests were carried out with cabbage looper larvae only, but no contact or ingested toxicity was detected. Our results suggest that oregonin at levels found in red alder leaves early in the growing season may contribute to protecting red alder from leaf-eating insects.

Application of amendments for the phytoremediation of a former mine technosol by endemic pioneer species: alder and birch seedlings.

Metal(loid) pollution of soils has important negative effects on the environment and human health. For the rehabilitation of these soils, some eco-innovative strategies, such as phytoremediation, could be chosen. This practice could establish a plant cover to reduce the toxicity of the pollutants and stabilize the soil, preventing soil erosion and water leaching; this technique is called phytoremediation. For this, plants need to be tolerant to the pollutants present; thus, phytoremediation can have better outcomes if endemic species of the polluted area are used. Finally, to further improve phytoremediation success, amendments can be applied to ameliorate soil conditions. Different amendments can be used, such as biochar, a good metal(loid) immobilizer, compost, a nutrient-rich product and iron sulfate, an efficient arsenic immobilizer. These amendments can either be applied alone or combined for further positive effects. In this context, a mesocosm experiment was performed to study the effects of three amendments, biochar, compost and iron sulfate, applied alone or combined to a former mine technosol, on the soil properties and the phytoremediation potential of two endemic species, Alnus sp. and Betula sp. Results showed that the different amendments reduced soil acidity and decreased metal(loid) mobility, thus improving plant growth. Both species were able to grow on the amended technosols, but alder seedlings had a much higher growth compared to birch seedlings. Finally, the combination of compost with biochar and/or iron sulfate and the establishment of endemic alder plants could be a solution to rehabilitate a former mine technosol.

Can Siberian alder N-fixation offset N-loss after severe fire? Quantifying post-fire Siberian alder distribution, growth, and N-fixation in boreal Alaska.

Fire severity affects both ecosystem N-loss and post-fire N-balance. Climate change is altering the fire regime of interior Alaska, although the effects on Siberian alder (Alnus viridis ssp. fruticosa) annual N-fixation input (kg N ha-1 yr-1) and ecosystem N-balance are largely unknown. We established 263 study plots across two burn scars within the Yukon-Tanana Uplands ecoregion of interior Alaska. Siberian alder N-input was quantified by post-fire age, fire severity, and stand type. We modeled the components of Siberian alder N-input using environmental variables and fire severity within and across burn scars and estimated post-fire N-balance using N-loss (volatilized N) and N-gain [biological N-fixation and atmospheric deposition]. Mean nodule-level N-fixation rate was 70% higher 11-years post-fire (12.88 ± 1.18 µmol N g-1 hr-1) than 40-years post-fire (7.58 ± 0.59 µmol N g-1 hr-1). Structural equation modeling indicated that fire severity had a negative effect on Siberian alder density, but a positive effect on live nodule biomass (g nodule m-2 plant-1). Post-fire Siberian alder N-input was highest in 11-year old moderately burned deciduous stands (11.53 ± 0.22 kg N ha-1 yr-1), and lowest in 11-year old stands that converted from black spruce to deciduous dominance after severe fire (0.06 ± 0.003 kg N ha-1 yr-1). Over a 138-year fire return interval, N-gains in converted black spruce stands are estimated to offset 15% of volatilized N, whereas N-gains in burned deciduous stands likely exceed volatilized N by an order of magnitude. High Siberian alder density and nodule biomass drives N-input in burned deciduous stands, while low N-fixer density (including Siberian alder) limits N-input in high severity black spruce stands not underlain by permafrost. A severe fire regime that converts black spruce stands to deciduous dominance without alder recruitment may induce progressive N-losses which alter boreal forest ecosystem patterns and processes.

Nitrogen-fixing red alder trees tap rock-derived nutrients.

Symbiotic nitrogen (N)-fixing trees supply significant N inputs to forest ecosystems, leading to increased soil fertility, forest growth, and carbon storage. Rapid growth and stoichiometric constraints of N fixers also create high demands for rock-derived nutrients such as phosphorus (P), while excess fixed N can generate acidity and accelerate leaching of rock-derived nutrients such as calcium (Ca). This ability of N-fixing trees to accelerate cycles of Ca, P, and other rock-derived nutrients has fostered speculation of a direct link between N fixation and mineral weathering in terrestrial ecosystems. However, field evidence that N-fixing trees have enhanced access to rock-derived nutrients is lacking. Here we use strontium (Sr) isotopes as a tracer of nutrient sources in a mixed-species temperate rainforest to show that N-fixing trees access more rock-derived nutrients than nonfixing trees. The N-fixing tree red alder (Alnus rubra), on average, took up 8 to 18% more rock-derived Sr than five co-occurring nonfixing tree species, including two with high requirements for rock-derived nutrients. The increased access to rock-derived nutrients occurred despite spatial variation in community-wide Sr sources across the forest, and only N fixers had foliar Sr isotopes that differed significantly from soil exchangeable pools. We calculate that increased uptake of rock-derived nutrients by N-fixing alder requires a 64% increase in weathering supply of nutrients over nonfixing trees. These findings provide direct evidence that an N-fixing tree species can also accelerate nutrient inputs from rock weathering, thus increasing supplies of multiple nutrients that limit carbon uptake and storage in forest ecosystems.

Leaf shedding increases the photosynthetic rate of the canopy in N2-fixing and non-N2-fixing woody species.

It has long been hypothesized that timing of leaf shedding is critical for plant fitness but there is little experimental evidence to support the hypothesis. According to an optimality theory, shedding of old leaves increases canopy photosynthesis despite some nitrogen (N) being lost as litterfall, when the ratio of daily photosynthesis to leaf N (N-use efficiency, ε) in old leaves, expressed as a fraction of ε in new leaves, becomes lower than the fraction of leaf N that is resorbed before shedding (RN). This was shown to be true for N-poor plants but not for N-rich plants in a pot experiment; however, the use of planting pots imposes a variety of physical, chemical and biological constraints that could change the experimental results. Here we conducted a 3-year field survey in a cool temperate deciduous forest to examine whether Alnus sieboldiana Matsum. (N2-fixing) and Carpinus tschonoskii Maxim. (non-N2-fixing) shed their leaves to increase canopy photosynthesis in accord with the above criterion. These species often grow sympatrically and were chosen as representatives of N-rich and N-poor plants, respectively. Overall, daily photosynthesis decreased with leaf age, accompanied by small changes in leaf N, resulting in a decrease in ε. In both species, ε of leaves at shedding expressed as a fraction of ε in new leaves was nearly equal to RN in all years, implying that the old leaves were shed to increase canopy photosynthesis. Our results, together with those of previous field surveys, suggested that the timing of leaf shedding is explained by N use in maximizing canopy photosynthesis across broad groups of species.

Host plant frequency and secondary metabolites are concurrently associated with insect herbivory in a dominant riparian tree.

Herbivory is strongly influenced by different sources of plant variation, from traits such as secondary metabolites to features associated with population- and community-level variation. However, most studies have assessed the influence of these drivers in isolation. We conducted a large-scale study to evaluate the associations between multiple types of plant-based variation and insect leaf herbivory in alder ( Alnus glutinosa) trees sampled in riparian forests throughout northwestern Spain. We assessed the associations between insect leaf herbivory and alder mean production of leaf secondary metabolites (phenolic compounds), variation among neighbouring alder trees in leaf phenolics and community-related features including alder relative size and frequency and tree species phylogenetic diversity. Structural equation modelling indicated that increasing concentrations of alder leaf flavonoids (but not other types of phenolic compounds) and increasing variation in phenolics among neighbouring alders were both significantly negatively associated with herbivory. In addition, increasing relative frequency of alder was positively associated with leaf damage, whereas the size of alders relative to other trees and phylogenetic diversity were not significantly associated with herbivory. These results demonstrate the concurrent and independent influences of different sources of plant-based variation on insect herbivory and argue for further future work simultaneously addressing multiple plant-based bottom-up controls.

Alder, Nitrogen, and Lake Ecology: Terrestrial-Aquatic Linkages in the Postglacial History of Lone Spruce Pond, Southwestern Alaska.

Diatoms, combined with a multiproxy study of lake sediments (organic matter, N, δ15N, δ13C, biogenic silica, grain size, Cladocera and chironomids, Alnus pollen) from Lone Spruce Pond, Alaska detail the late-glacial to Holocene history of the lake and its response to regional climate and landscape change over the last 14.5 cal ka BP. We show that the immigration of alder (Alnus viridis) in the early Holocene marks the rise of available reactive nitrogen (Nr) in the lake as well as the establishment of a primarily planktonic diatom community. The later establishment of diatom Discostella stelligera is coupled to a rise of sedimentary δ15N, indicating diminished competition for this nutrient. This terrestrial-aquatic linkage demonstrates how profoundly vegetation may affect soil geochemistry, lake development, and lake ecology over millennial timescales. Furthermore, the response of the diatom community to strengthened stratification and N levels in the past confirms the sensitivity of planktonic diatom communities to changing thermal and nutrient regimes. These past ecosystem dynamics serve as an analogue for the nature of threshold-type ecological responses to current climate change and atmospheric nitrogen (Nr) deposition, but also for the larger changes we should anticipate under future climate, pollution, and vegetation succession scenarios in high-latitude and high-elevation regions.

Nitrogen isotope fractionation during N uptake via arbuscular mycorrhizal and ectomycorrhizal fungi into grey alder.

Arbuscular mycorrhizal (AM) and ectomycorrhizal (ECM) fungi affect plant nitrogen (N) dynamics. Plant N isotope patterns have been used to characterise the contribution of ECM fungi to plant N uptake. By quantifying and comparing the effects of an AM and an ECM fungus on growth, N uptake and isotopic composition of one host plant grown at different relative N supply levels, the aim of this study was to improve the mechanistic understanding of natural 15N abundance patterns in mycorrhizal plants and their underlying causes. Grey alders were inoculated with one ECM fungus or one AM fungus or left non-mycorrhizal. Plants were grown under semi-hydroponic conditions and were supplied with three rates of relative N supply ranging from deficient to luxurious. Neither mycorrhizal fungus increased plant growth or N uptake. AM root colonisation had no effect on whole plant δ15N and decreased foliar δ 15N only under N deficiency. The roots of these plants were 15N-enriched. ECM root colonisation consistently decreased foliar and whole plant δ15N. It is concluded, that both mycorrhizal fungi contributed to plant N uptake into the shoot. Nitrogen isotope fractionation during N assimilation and transformations in fungal mycelia is suggested to have resulted in plants receiving 15N-depleted N via the mycorrhizal uptake pathways. Negative mycorrhizal growth effects are explained by symbiotic resource trade on carbon and N and decreased direct plant N uptake.

Temperature and substrate chemistry as major drivers of interregional variability of leaf microbial decomposition and cellulolytic activity in headwater streams.

Abiotic factors, substrate chemistry and decomposers community composition are primary drivers of leaf litter decomposition. In soil, much of the variation in litter decomposition is explained by climate and substrate chemistry, but with a significant contribution of the specialisation of decomposer communities to degrade specific substrates (home-field advantage, HFA). In streams, however, HFA effects on litter decomposition have not been explicitly tested. We evaluated responses of microbial decomposition and ß-glucosidase activity to abiotic factors, substrate and decomposer assemblages, using a reciprocal litter transplant experiment: 'ecosystem type' (mountain vs lowland streams) × 'litter chemistry' (alder vs reed). Temperature, pH and ionic concentration were higher in lowland streams. Decomposition for both species was faster in lowland streams. Decomposition of reed was more accelerated in lowland compared with mountain streams than that of alder, suggesting higher temperature sensitivity of decomposition in reed. Q10 (5°C-15°C) values of ß-glucosidase activity were over 2. The alkaline pH and high ionic concentration of lowland streams depleted enzyme activity. We found similar relationships of decomposition or enzyme activity with abiotic factors for both species, suggesting limited support to the HFA hypothesis. Overall, our results suggest a prime role of temperature interacting with substrate chemistry on litter decomposition.

Physiological effects of major up-regulated Alnus glutinosa peptides on Frankia sp. ACN14a.

Alnus glutinosa has been shown previously to synthesize, in response to nodulation by Frankia sp. ACN14a, an array of peptides called Alnus symbiotic up-regulated peptides (ASUPs). In a previous study one peptide (Ag5) was shown to bind to Frankia nitrogen-fixing vesicles and to modify their porosity. Here we analyse four other ASUPs, alongside Ag5, to determine whether they have different physiological effects on in vitro grown Frankia sp. ACN14a. The five studied peptides were shown to have different effects on nitrogen fixation, respiration, growth, the release of ions and amino acids, as well as on cell clumping and cell lysis. The mRNA abundance for all five peptides was quantified in symbiotic nodules and one (Ag11) was found to be more abundant in the meristem part of the nodule. These findings point to some peptides having complementary effects on Frankia cells.

Effect of Uranium and Thorium Radionuclides on Biochemical Characteristics of Duschekia fruticosa in "Soil-Plant" System.

The biochemical characteristics of Duschekiafruticosa, grown for a long time under a variety of exposure doses of natural background radiation (up to 150 µR/h) was studied. Uranium was found to make the dominant contribution to the y-background exposure doses. The pH-values and the content of organic matter in soils within the surveyed territory remained unchanged. Accumulation of radionuclides of uranium and thorium in the "soil-plant" system was studied. It is shown for the D. fruticosa that U and Th uptake decreased with y-background increasing. Study of anti-free radical and anti-peroxide cells' protection system indicated a balanced activity of prooxidant-antioxidant systems in the cells of the D. fruticosa leaves. The combined effect of incorporated uranium and thorium is accompanied by a significant increase in chlorophyll content in D. fruticosa.

High resource-capture and -use efficiency, and effective antioxidant protection contribute to the invasiveness of Alnus formosana plants.

To investigate the traits contributing to the invasiveness of Alnus formosana and the mechanisms underlying its invasiveness, we compared A. formosana with its native congener (Alnus cremastogyne) under three light treatments (13%, 56%, and 100%). The consistently higher plant height, total leaf area, light-saturated photosynthetic rate (A(max)), light saturation point (LSP), light compensation point (LCP), respiration efficiency (RE), and non-photochemical quenching coefficient (NPQ) but lower root mass fraction (RMF) and specific leaf area (SLA) of the invader than of its native congener contributed to the higher RGR and total biomass of A. formosana across light regimes. The total biomass and RGR of the invader increased markedly with increased RMF, A(max), LSP, LCP, RE, stomatal conductance (G(s)) and total leaf area. Furthermore, compared with the native species, the higher plasticity index in plant height, RMF, leaf mass fraction (LMF), SMF, SLA, A(max) and dark respiration rate (R(d)) within the range of total light contributed to the higher performance of the invader. In addition, the activities of antioxidant enzymes were higher in the invader compared to the native, contributing to its invasion success under high/low light via photoprotection. With a decrease in light level, superoxide dismutase (SOD) and catalase (CAT) activities increased significantly, whereas total carotenoid (Car) and total chlorophyll (Chl) decreased; ascorbate peroxidase (APX) and glutathione reductase (GR) activities remained unchanged. These responses may help the invader to spread and invade a wide range of habitats and form dense monocultures, displacing native plant species. The results suggest that both resource capture-related traits (morphological and photosynthetic) and adaptation-related traits (antioxidant protection) contribute to the competitive advantage of the invader.

Ecological consequences of the expansion of N2-fixing plants in cold biomes.

Research in warm-climate biomes has shown that invasion by symbiotic dinitrogen (N2)-fixing plants can transform ecosystems in ways analogous to the transformations observed as a consequence of anthropogenic, atmospheric nitrogen (N) deposition: declines in biodiversity, soil acidification, and alterations to carbon and nutrient cycling, including increased N losses through nitrate leaching and emissions of the powerful greenhouse gas nitrous oxide (N2O). Here, we used literature review and case study approaches to assess the evidence for similar transformations in cold-climate ecosystems of the boreal, subarctic and upper montane-temperate life zones. Our assessment focuses on the plant genera Lupinus and Alnus, which have become invasive largely as a consequence of deliberate introductions and/or reduced land management. These cold biomes are commonly located in remote areas with low anthropogenic N inputs, and the environmental impacts of N2-fixer invasion appear to be as severe as those from anthropogenic N deposition in highly N polluted areas. Hence, inputs of N from N2 fixation can affect ecosystems as dramatically or even more strongly than N inputs from atmospheric deposition, and biomes in cold climates represent no exception with regard to the risk of being invaded by N2-fixing species. In particular, the cold biomes studied here show both a strong potential to be transformed by N2-fixing plants and a rapid subsequent saturation in the ecosystem's capacity to retain N. Therefore, analogous to increases in N deposition, N2-fixing plant invasions must be deemed significant threats to biodiversity and to environmental quality.

Temperature affects leaf litter decomposition in low-order forest streams: field and microcosm approaches.

Despite predicted global warming, the temperature effects on headwater stream functioning are poorly understood. We studied these effects on microbial-mediated leaf decomposition and the performance of associated aquatic hyphomycete assemblages. Alder leaves were incubated in three streams differing in winter water temperature. Simultaneously, in laboratory, leaf discs conditioned in these streams were incubated at 5, 10 and 15 °C. We determined mass loss, leaf N and sporulation rate and diversity of aquatic hyphomycete communities. In the field, decomposition rate correlated positively with temperature. Decomposition rate and leaf N presented a positive trend with dissolved nutrients, suggesting that temperature was not the only factor determining the process velocity. Under controlled conditions, it was confirmed that decomposition rate and leaf N were positively correlated with temperature, leaves from the coldest stream responding most clearly. Sporulation rate correlated positively with temperature after 9 days of incubation, but negatively after 18 and 27 days. Temperature rise affected negatively the sporulating fungi richness and diversity only in the material from the coldest stream. Our results suggest that temperature is an important factor determining leaf processing and aquatic hyphomycete assemblages and that composition and activity of fungal communities adapted to cold environments could be more affected by temperature rises. Highlight: Leaf decomposition rate and associated fungal communities respond to temperature shifts in headwater streams.

From tolerance to acute metabolic deregulation: contribution of proteomics to dig into the molecular response of alder species under a polymetallic exposure.

Alnus spp. are actinorhizal trees commonly found in wet habitats and able to grow effectively in soil slightly contaminated with metal trace- elements. Two clones belonging to two Alnus species, namely, A. incana and A. glutinosa, were grown in hydroponics and exposed for 9 weeks to a Cd + Ni + Zn polymetallic constraint. Although responding by a similar decrease in total biomass production, the proteomic analysis associated with the study of various biochemical parameters including carbohydrate and mineral analyses revealed that the two clones have a distinct stress-responsive behavior. All parameters indicated that the roots, the organ in direct contact with the media, are more affected than the leaves. In fact, in A. glutinosa the response was almost completely confined to the roots, whereas many proteins change significantly in the roots and in the leaves of the treated A. incana. In both clones, the changes affected a broad range of metabolic processes such as redox regulation and energy metabolism and induced the production of pathogenesis-related proteins. In particular, changes in the accumulation of bacterial proteins that were not identified as coming from the known symbionts of Alnus were reported. Further investigation should be performed to identify their origin and exact role in the plant response to the polymetallic exposure tested here.

Effects of riparian plant diversity loss on aquatic microbial decomposers become more pronounced with increasing time.

We examined the potential long-term impacts of riparian plant diversity loss on diversity and activity of aquatic microbial decomposers. Microbial assemblages were obtained in a mixed-forest stream by immersion of mesh bags containing three leaf species (alder, oak and eucalyptus), commonly found in riparian corridors of Iberian streams. Simulation of species loss was done in microcosms by including a set of all leaf species, retrieved from the stream, and non-colonized leaves of three, two or one leaf species. Leaves were renewed every month throughout six months, and microbial inoculum was ensured by a set of colonized leaves from the previous month. Microbial diversity, leaf mass loss and fungal biomass were assessed at the second and sixth months after plant species loss. Molecular diversity of fungi and bacteria, as the total number of operational taxonomic units per leaf diversity treatment, decreased with leaf diversity loss. Fungal biomass tended to decrease linearly with leaf species loss on oak and eucalyptus, suggesting more pronounced effects of leaf diversity on lower quality leaves. Decomposition of alder and eucalyptus leaves was affected by leaf species identity, mainly after longer times following diversity loss. Leaf decomposition of alder decreased when mixed with eucalyptus, while decomposition of eucalyptus decreased in mixtures with oak. Results suggest that the effects of leaf diversity on microbial decomposers depended on leaf species number and also on which species were lost from the system, especially after longer times. This may have implications for the management of riparian forests to maintain stream ecosystem functioning.