Contribution of microbial activity and vegetation cover to the spatial distribution of soil respiration in mountains.

The patterns of change in bioclimatic conditions determine the vegetation cover and soil properties along the altitudinal gradient. Together, these factors control the spatial variability of soil respiration (RS) in mountainous areas. The underlying mechanisms, which are poorly understood, shape the resulting surface CO2 flux in these ecosystems. We aimed to investigate the spatial variability of RS and its drivers on the northeastern slope of the Northwest Caucasus Mountains, Russia (1,260-2,480 m a.s.l.), in mixed, fir, and deciduous forests, as well as subalpine and alpine meadows. RS was measured simultaneously in each ecosystem at 12 randomly distributed points using the closed static chamber technique. After the measurements, topsoil samples (0-10 cm) were collected under each chamber (n = 60). Several soil physicochemical, microbial, and vegetation indices were assessed as potential drivers of RS. We tested two hypotheses: (i) the spatial variability of RS is higher in forests than in grasslands; and (ii) the spatial variability of RS in forests is mainly due to soil microbial activity, whereas in grasslands, it is mainly due to vegetation characteristics. Unexpectedly, RS variability was lower in forests than in grasslands, ranging from 1.3-6.5 versus 3.4-12.7 µmol CO2 m-1 s-1, respectively. Spatial variability of RS in forests was related to microbial functioning through chitinase activity (50% explained variance), whereas in grasslands it was related to vegetation structure, namely graminoid abundance (27% explained variance). Apparently, the chitinase dependence of RS variability in forests may be related to soil N limitation. This was confirmed by low N content and high C:N ratio compared to grassland soils. The greater sensitivity of grassland RS to vegetation structure may be related to the essential root C allocation for some grasses. Thus, the first hypothesis concerning the higher spatial variability of RS in forests than in grasslands was not confirmed, whereas the second hypothesis concerning the crucial role of soil microorganisms in forests and vegetation in grasslands as drivers of RS spatial variability was confirmed.

Functional responses of two Mediterranean pine species in an ozone Free-Air Controlled Exposure (FACE) experiment.

Effects of the phytotoxic and widespread ozone (O3) pollution may be species specific, but knowledge on Mediterranean conifer responses to long-term realistic exposure is still limited. We examined responses regarding to photosynthesis, needle biochemical stress markers and carbon and nitrogen (N) isotopes of two Mediterranean pine species (Pinus halepensis Mill. and Pinus pinea L.). Seedlings were grown in a Free-Air Controlled Exposure experiment with three levels of O3 (ambient air, AA [38.7 p.p.b. as daily average]; 1.5 × AA and 2.0 × AA) during the growing season (May-October 2019). In P. halepensis, O3 caused a significant decrease in the photosynthetic rate, which was mainly due to a reduction of both stomatal and mesophyll diffusion conductance to CO2. Isotopic analyses indicated a cumulative or memory effect of O3 exposure on this species, as the negative effects were highlighted only in the late growing season in association with a reduced biochemical defense capacity. On the other hand, there was no clear effect of O3 on photosynthesis in P. pinea. However, this species showed enhanced N allocation to leaves to compensate for reduced photosynthetic N- use efficiency. We conclude that functional responses to O3 are different between the two species determining that P. halepensis with thin needles was relatively sensitive to O3, while P. pinea with thicker needles was more resistant due to a potentially low O3 load per unit mass of mesophyll cells, which may affect species-specific resilience in O3-polluted Mediterranean pine forests.

Diversity of Microbial Communities, PAHs, and Metals in Road and Leaf Dust of Functional Zones of Moscow and Murmansk.

The impact of geographical factors, functional zoning, and biotope type on the diversity of microbial communities and chemical components in the dust of urban ecosystems was studied. Comprehensive analyses of bacterial and fungal communities, polycyclic aromatic hydrocarbons (PAHs), and metals in road and leaf dust in three urban zones of Murmansk and Moscow with contrasting anthropogenic load were conducted. We found that the structure of bacterial communities affected the functional zoning of the city, biotope type, and geographical components. Fungal communities were instead impacted only by biotope type. Our findings revealed that the structure of fungal communities was mostly impacted by PAHs whereas bacterial communities were sensitive to metals. Bacteria of the genus Sphingomonas in road and leaf dust as indicators of the ecological state of the urban ecosystems were proposed.

Polycyclic Aromatic Hydrocarbon-Degrading Bacteria in Three Different Functional Zones of the Cities of Moscow and Murmansk.

We performed a comparative study of the total bacterial communities and communities of cultivable polycyclic aromatic hydrocarbons (PAH)-degrading bacteria in different functional zones of Moscow and Murmansk that were formed under the influence of the PAH composition in road and leaf dust. The PAHs were determined by high-performance liquid chromatography (HPLC); the bacterial communities' diversity was assessed by metabarcoding. The degraders were isolated by their direct plating on a medium with the PAHs. The PAH total quantity declined in the leaf dust from the traffic to the recreational zone. For the road dust, a negative gradient with pollution was observed for Rhodococcus and Acinetobacter degraders and for their relative abundance in the microbiome for the functional zones of Moscow. The opposite effect was observed in the Murmansk leaf dust for the Rothia and Pseudomonas degraders and in the Moscow road dust for Microbacterium. The PCA and linear regression analyses showed that the Micrococcus degraders in the dust were sensitive to anthropogenic pollution, so they can be used as a tool for monitoring anthropogenic changes in the biosphere. The data on the degraders' and microbial communities' diversity suggest that minor degrading strains can play a key role in PAH degradation.

Consequences of Grazing Cessation for Soil Environment and Vegetation in a Subalpine Grassland Ecosystem.

Areas covered by seminatural grasslands have been in constant decline for decades in Europe. This trend is particularly strong for mountain territories, where such traditional agricultural practices as cattle grazing are no longer economically feasible. This study was conducted in the subalpine pasture of Cinte Tesino (TN, Italy), where local farmers have applied the following different management strategies: shorter and longer grazing durations during the season and a complete abandonment for the last 15 years. We aimed to study how these different management strategies impact the functioning and diversity of vegetation and the chemical and biological characteristics of the soil. Species richness was higher in plots subjected to longer grazing with a prevalence of D. caespitosa in terms of biomass share. A decline in species richness in abandoned plots was accompanied by an increase in the share of other graminoids in collected biomass. A concomitant increase in leaf N concentration and light availability in grazed plots resulted in higher photosynthetic efficiency in some species, as revealed by the δ13C of plant tissues. Soils under grazing were characterised by a higher concentration of total and extractable N, almost doubled microbial biomass C and increased extracellular enzymes activity, evidencing nutrient cycling mobilization. While the microbial pool was characterised by lower mineralization rates, C was lost from the soil with 15 years of abandonment. The longer grazing season demonstrated to be the most beneficial, promoting species richness, C accumulation and better soil microbial functioning. A change in soil pH from strongly acidic to moderately acidic with longer grazing is likely one of the important factors adding to the success in the functioning of primary producers and decomposers in this site.

Phylloplane Biodiversity and Activity in the City at Different Distances from the Traffic Pollution Source.

The phylloplane is an integrated part of green infrastructure which interacts with plant health. Taxonomic characterization of the phylloplane with the aim to link it to ecosystem functioning under anthropogenic pressure is not sufficient because only active microorganisms drive biochemical processes. Activity of the phylloplane remains largely overlooked. We aimed to study the interactions among the biological characteristics of the phylloplane: taxonomic diversity, functional diversity and activity, and the pollution grade. Leaves of Betula pendula were sampled in Moscow at increasing distances from the road. For determination of phylloplane activity and functional diversity, a MicroResp tool was utilized. Taxonomic diversity of the phylloplane was assessed with a combination of microorganism cultivation and molecular techniques. Increase of anthropogenic load resulted in higher microbial respiration and lower DNA amount, which could be viewed as relative inefficiency of phylloplane functioning in comparison to less contaminated areas. Taxonomic diversity declined with road vicinity, similar to the functional diversity pattern. The content of Zn in leaf dust better explained the variation in phylloplane activity and the amount of DNA. Functional diversity was linked to variation in nutrient content. The fraction of pathogenic fungi of the phylloplane was not correlated with any of the studied elements, while it was significantly high at the roadsides. The bacterial classes Gammaproteobacteria and Cytophagia, as well as the Dothideomycetes class of fungi, are exposed to the maximal effect of distance from the highway. This study demonstrated the sensitivity of the phylloplane to road vicinity, which combines the effects of contaminants (mainly Zn according to this study) and potential stressful air microclimatic conditions (e.g., low relative air humidity, high temperature, and UV level). Microbial activity and taxonomic diversity of the phylloplane could be considered as an additional tool for bioindication.

Carbon allocation strategies and water uptake in young grafted and own-rooted hazelnut (Corylus avellana L.) cultivars.

In this study, grafted and own-rooted young hazelnut plants of three high-quality cultivars were cultivated in Central Italy to investigate possible differences in growth, fruit and flower production, and physiological processes encompassing water uptake, photosynthetic variables and non-structural carbohydrate allocation. Stable isotopes and photosynthetic measurements were used to study carbon and water fluxes in plants. For the first time, an ecophysiological study was carried out to understand the seasonal growth dynamics of grafted plants in comparison with own-rooted plants. The own-rooted hazelnuts showed rapid above-ground development with large canopy volume, high amount of sprouts and earlier yield. The grafted plants showed greater below-ground development with lower canopy volumes and lower yield. However, later, the higher growth rates of the canopy led these plants to achieve the same size as that of the own-rooted hazelnuts and to enter the fruit production phase. Different seasonal behaviour in root water uptake and leaf photosynthesis-related variables was detected between the two types of plants. The grafted plants showed root development that allowed deeper water uptake than that of the own-rooted hazelnuts. Moreover, the grafted plants were characterized by a higher accumulation of carbohydrate reserves in their root tissues and by higher stomatal reactivity, determining significant plasticity in response to seasonal thermal variations.

Winter's bite: beech trees survive complete defoliation due to spring late-frost damage by mobilizing old C reserves.

Late frost can destroy the photosynthetic apparatus of trees. We hypothesized that this can alter the normal cyclic dynamics of C-reserves in the wood. We measured soluble sugar concentrations and radiocarbon signatures (Δ14 C) of soluble nonstructural carbon (NSC) in woody tissues sampled from a Mediterranean beech forest that was completely defoliated by an exceptional late frost in 2016. We used the bomb radiocarbon approach to estimate the time elapsed since fixation of mobilized soluble sugars. During the leafless period after the frost event, soluble sugar concentrations declined sharply while Δ14 C of NSC increased. This can be explained by the lack of fresh assimilate supply and a mobilization of C from reserve pools. Soluble NSC became increasingly older during the leafless period, with a maximum average age of 5 yr from samples collected 27 d before canopy recovery. Following leaf re-growth, soluble sugar concentrations increased and Δ14 C of soluble NSC decreased, indicating the allocation of new assimilates to the stem soluble sugars pool. These data highlight that beech trees rapidly mobilize reserve C to survive strong source-sink imbalances, for example due to late frost, and show that NSC is a key trait for tree resilience under global change.

Emission of constitutive isoprene, induced monoterpenes, and other volatiles under high temperatures in Eucalyptus camaldulensis: A 13 C labelling study.

Eucalypts are major emitters of biogenic volatile organic compounds (BVOCs), especially volatile isoprenoids. Emissions and incorporation of 13 C in BVOCs were measured in Eucalyptus camaldulensis branches exposed to rapid heat stress or progressive temperature increases, in order to detect both metabolic processes and their dynamics. Isoprene emission increased and photosynthesis decreased with temperatures rising from 30°C to 45°C, and an increasing percentage of unlabelled carbon was incorporated into isoprene in heat-stressed leaves. Intramolecular labelling was also incomplete in isoprene emitted by heat-stressed leaves, suggesting increasing contribution of respiratory (and possibly also photorespiratory) carbon. At temperature above 45°C, a drop of isoprene emission was mirrored by the appearance of unlabelled monoterpenes, green leaf volatiles, methanol, and ethanol, indicating that the emission of stored volatiles was mainly induced by cellular damage. Emission of partially labelled acetaldehyde was also observed at very high temperatures, suggesting a double source of carbon, with a large unlabelled component likely transported from roots and associated to the surge of transpiration at very high temperatures. Eucalypt plantations cover large areas worldwide, and our findings may dramatically change forecast and modelling of future BVOC emissions at planetary level, especially considering climate warming and frequent heat waves.

Effects of rain shortage on carbon allocation, pools and fluxes in a Mediterranean shrub ecosystem - a 13C labelling field study.

Hydrological cycle is expected to become the primary cause of ecosystem's degradation in near future under changing climate. Rain manipulation experiments under field conditions provide accurate picture on the responses of biotic processes to changed water availability for plants. A field experiment, mimicking expected changes in rain patterns, was established in a Mediterranean shrub community at Porto Conte, Italy, in 2001. In November 2011 Cistus monspeliensis, one of the dominating shrub species in the Mediterranean basin, was 13C labelled on plots subjected to extended rain shortage period and on control non manipulated plots. Carbon (C) allocation was traced by 13C dynamics in shoots, shoot-respired CO2, roots, microbial biomass, K2SO4-extractable C and CO2 respired from soil. Most of the recovered 13C (60%) was respired by shoots within 2weeks in control plots. In rain shortage treatment, 13C remained incorporated in aboveground plant parts. Residence time of 13C in leaves was longer under the rain shortage because less 13C was lost by shoot respiration and because 13C was re-allocated to leaves from woody tissues. The belowground C sink was weak (3-4% of recovered 13C) and independent on rain manipulation. Extended rain shortage promoted C exudation into rhizosphere soil in expense of roots. Together with lowered photosynthesis, this "save" economy of new C metabolites reduces the growing season under rain shortage resulting in decrease of shrub cover and C losses from the system on the long-term.

Carbon and nitrogen allocation strategy in Posidonia oceanica is altered by seawater acidification.

Rising atmospheric CO2 causes ocean acidification that represents one of the major ecological threats for marine biota. We tested the hypothesis that long-term exposure to increased CO2 level and acidification in a natural CO2 vent system alters carbon (C) and nitrogen (N) metabolism in Posidonia oceanica L. (Delile), affecting its resilience, or capability to restore the physiological homeostasis, and the nutritional quality of organic matter available for grazers. Seawater acidification decreased the C to N ratio in P. oceanica tissues and increased grazing rate, shoot density, leaf proteins and asparagine accumulation in rhizomes, while the maximum photochemical efficiency of photosystem II was unaffected. The 13C-dilution in both structural and non-structural C metabolites in the acidified site indicated quali-quantitative changes of C source and/or increased isotopic fractionation during C uptake and carboxylation associated with the higher CO2 level. The decreased C:N ratio in the acidified site suggests an increased N availability, leading to a greater storage of 15N-enriched compounds in rhizomes. The amount of the more dynamic C storage form, sucrose, decreased in rhizomes of the acidified site in response to the enhanced energy demand due to higher shoot recruitment and N compound synthesis, without affecting starch reserves. The ability to modulate the balance between stable and dynamic C reserves could represent a key ecophysiological mechanism for P. oceanica resilience under environmental perturbation. Finally, alteration in C and N dynamics promoted a positive contribution of this seagrass to the local food web.

Investigating the European beech (Fagus sylvatica L.) leaf characteristics along the vertical canopy profile: leaf structure, photosynthetic capacity, light energy dissipation and photoprotection mechanisms.

Forest functionality and productivity are directly related to canopy light interception and can be affected by potential damage from high irradiance. However, the mechanisms by which leaves adapt to the variable light environments along the multilayer canopy profile are still poorly known. We explored the leaf morphophysiological and metabolic responses to the natural light gradient in a pure European beech (Fagus sylvatica L.) forest at three different canopy heights (top, middle and bottom). Structural adjustment through light-dependent modifications in leaf mass per area was the reason for most of the variations in photosynthetic capacity. The different leaf morphology along the canopy influenced nitrogen (N) partitioning, water- and photosynthetic N-use efficiency, chlorophyll (Chl) fluorescence and quali-quantitative contents of photosynthetic pigments. The Chl a to Chl b ratio and the pool of xanthophyll-cycle pigments (VAZ) increased at the highest irradiance, as well as lutein and ß-carotene. The total pool of ascorbate and phenols was higher in leaves of the top and middle canopy layers when compared with the bottom layer, where the ascorbate peroxidase was relatively more activated. The non-photochemical quenching was strongly and positively related to the VAZ/(Chl a + b) ratio, while Chl a/Chl b was related to the photochemical efficiency of photosystem II. Along the multilayer canopy profile, the high energy dissipation capacity of leaves was correlated to an elevated redox potential of antioxidants. The middle layer gave the most relevant contribution to leaf area index and carboxylation capacity of the canopy. In conclusion, a complex interplay among structural, physiological and biochemical traits drives the dynamic leaf acclimation to the natural gradients of variable light environments along the tree canopy profile. The relevant differences observed in leaf traits within the canopy positions of the beech forest should be considered for improving estimation of carbon fluxes in multilayer canopy models of temperate forests.