Visually evoked local field potential changes in the caudate nucleus are remarkably more frequent in awake, behaving cats than in anaesthetized animals.

Previous results show that halothane gas anaesthesia has a suppressive effect on the visually evoked single-cell activities in the feline caudate nucleus (CN). In this study, we asked whether the low-frequency neuronal signals, the local field potentials (LFP) are also suppressed in the CN of anaesthetized animals.To answer this question, we compared the LFPs recorded from the CN of two halothane-anaesthetized (1.0%), paralyzed, and two awake, behaving cats during static and dynamic visual stimulation. The behaving animals were trained to perform a visual fixation task.Our results denoted a lower proportion of significant power changes to visual stimulation in the CN of the anesthetized cats in each frequency range (from delta to beta) of the LFPs, except gamma. These differences in power changes were more obvious in static visual stimulation, but still, remarkable differences were found in dynamic stimulation, too. The largest differences were found in the alpha and beta frequency bands for static stimulation. Concerning dynamic stimulation, the differences were the biggest in the theta, alpha and beta bands.Similar to the single-cell activities, remarkable differences were found between the visually evoked LFP changes in the CN of the anaesthetized, paralyzed and awake, behaving cats. The halothane gas anaesthesia and the immobilization suppressed the significant LFP power alterations in the CN to both static and dynamic stimulation. These results suggest the priority of the application of behaving animals even in the analysis of the visually evoked low-frequency electric signals, the LFPs recorded from the CN.

Automated preprocessing of 64 channel electroenchephalograms recorded by biosemi instruments.

Preprocessing is a mandatory step in electroencephalogram (EEG) signal analysis. Overcoming challenges posed by high noise levels and substantial amplitude artifacts, such as blink-induced electrooculogram (EOG) and muscle-related electromyogram (EMG) interference, is imperative. The signal-to-noise ratio significantly influences the reliability and statistical significance of subsequent analyses. Existing referencing approaches employed in multi-card systems, like using a single electrode or averaging across multiple electrodes, fall short in this respect. In this article, we introduce an innovative referencing method tailored to multi-card instruments, enhancing signal fidelity and analysis outcomes. Our proposed signal processing loop not only mitigates blink-related artifacts but also accurately identifies muscle activity. This work contributes to advancing EEG analysis by providing a robust solution for artifact removal and enhancing data integrity.•Removes blink•Marks muscle activity•Re-references with design specific enhancements.

PADAPT 1.0 - the Pannonian Dataset of Plant Traits.

The existing plant trait databases' applicability is limited for studies dealing with the flora and vegetation of the eastern and central part of Europe and for large-scale comparisons across regions, mostly because their geographical data coverage is limited and they incorporate records from several different sources, often from regions with markedly different climatic conditions. These problems motivated the compilation of a regional dataset for the flora of the Pannonian region (Eastern Central Europe). PADAPT, the Pannonian Dataset of Plant Traits relies on regional data sources and collates data on 54 traits and attributes of the plant species of the Pannonian region. The current version covers approximately 90% of the species of the region and consists of 126,337 records on 2745 taxa. By including species of the eastern part of Europe not covered by other databases, PADAPT can facilitate studying the flora and vegetation of the eastern part of the continent. Although data coverage is far from complete, PADAPT meets the longstanding need for a regional database of the Pannonian flora.

How to not trade water for carbon with tree planting in water-limited temperate biomes?

The most widespread nature-based solution for mitigating climate change is tree planting. When realized as forest restoration in historically forested biomes, it can efficiently contribute to the sequestration of atmospheric carbon and can also entail significant biodiversity and ecosystem service benefits. Conversely, tree planting in naturally open biomes can have adverse effects, of which water shortage due to increased evapotranspiration is among the most alarming ones. Here we assessed how soil texture affects the strength of the trade-off between tree cover and water balance in the forest-steppe biome, where the global pressure for afforestation is threatening with increasing tree cover above historical levels. Here we monitored vertical soil moisture dynamics in four stands in each of the most common forest types of lowland Hungary on well-drained, sandy (natural poplar groves, and Robinia and pine plantations) and on poorly drained, silty-clayey soils (natural oak stands and Robinia plantations), and neighboring grasslands. We found that forests on sand retain moisture in the topsoil (approx. 20 cm) throughout the year, but a thick dry layer develops below that during the vegetation period, significantly impeding groundwater recharge. Neighboring sandy grasslands showed an opposite pattern, with often dry topsoil but intact moisture reserves below, allowing deep percolation. In contrast, forests on silty-clayey soils did not desiccate lower soil layers compared neighboring grasslands, which in turn showed moisture patterns similar to sandy grasslands. We conclude that, in water-limited temperate biomes where landscape-wide water regime depends on deep percolation, soil texture should drive the spatial allocation of tree-based climate mitigation efforts. On sand, the establishment of new forests should be kept to a minimum and grassland restoration should be preferred. The trade-off between water and carbon is less pronounced on silty-clayey soils, making forest patches and wooded rangelands viable targets for both climate mitigation and ecosystem restoration.

Co-seeding grasses and forbs supports restoration of species-rich grasslands and improves weed control in ex-arable land.

Sowing is widely used for the restoration of species-rich grasslands but still there are knowledge gaps regarding the most suitable application of different seed mixtures. We tested the effect of seed mixtures application timing on the establishment of sown forbs and weed control. 36 experimental plots with nine sowing treatments were established in an abandoned cropland in Hungary. Grass-seeds, diverse forb seed mixture and the combination of the two were applied: diverse forb mixture was sown simultaneously or 1, 2 or 3 years after grass sowing, in plots sown previously with grass or in empty plots (fallows). All sowing treatments supported the rapid establishment of the sown species in large cover and hampered weed encroachment. Forbs performed better when sown into fallows than in grass-matrix and forbs establishment was worse in older fallows than in younger ones. Grasses expressed a strong priority effect, especially when forbs were sown at least two years later than grasses. We also investigated the relation between seed germinability, weather parameters and establishment success. Germination rate in the greenhouse could not predict the establishment success of forbs in the field and showed great differences between years, hence we recommend sowing target forbs in multiple years.

Patch and matrix characteristics determine the outcome of ecosystem engineering by mole rats in dry grasslands.

Background: Burrowing mammals are important ecosystem engineers, especially in open ecosystems where they create patches that differ from the surrounding matrix in their structure or ecosystem functions. Methods: We evaluated the fine-scale effects of a subterranean ecosystem engineer, the Lesser blind mole rat on the vegetation composition of sandy dry grasslands in Hungary. In this model system we tested whether the characteristics of the patch (mound size) and the matrix (total vegetation cover in the undisturbed grassland) influence the structural and functional contrasts between the mounds and the undisturbed grasslands. We sampled the vegetation of 80 mounds and 80 undisturbed grassland plots in four sites, where we recorded the total vegetation cover, and the occurrence and cover of each vascular plant species. We used two proxies to characterise the patches (mounds) and the matrix (undisturbed grassland): we measured the perimeter of the mounds and estimated the total vegetation cover of the undisturbed grasslands. First, we compared the vegetation characteristics of the mounds and the surrounding grasslands with general linear models. Second, we characterised the contrasts between the mounds and the undisturbed grassland by relative response indices (RRIs) of the vegetation characteristics studied in the first step. Results: Species composition of the vegetation of the mounds and undisturbed grasslands was well separated in three out of the four study sites. Mounds were characterised by lower vegetation cover, lower cover of perennial graminoids, and higher diversity, and evenness compared to undisturbed grasslands. The contrast in vegetation cover between mounds and undisturbed grasslands increased with decreasing patch size. Increasing vegetation cover in the matrix grasslands increased the contrasts between the mounds and undisturbed grasslands in terms of total cover, perennial graminoid cover, diversity, and evenness. Our results suggest that mole rat mounds provide improved establishment conditions for subordinate species, because they are larger than other types of natural gaps and are characterised by less intense belowground competition. The ecosystem engineering effect, i.e., the contrast between the patches and the matrix was the largest in the more closed grasslands.

Visual consequent stimulus complexity affects performance in audiovisual associative learning.

In associative learning (AL), cues and/or outcome events are coupled together. AL is typically tested in visual learning paradigms. Recently, our group developed various AL tests based on the Rutgers Acquired Equivalence Test (RAET), both visual and audiovisual, keeping the structure and logic of RAET but with different stimuli. In this study, 55 volunteers were tested in two of our audiovisual tests, SoundFace (SF) and SoundPolygon (SP). The antecedent stimuli in both tests are sounds, and the consequent stimuli are images. The consequents in SF are cartoon faces, while in SP, they are simple geometric shapes. The aim was to test how the complexity of the applied consequent stimuli influences performance regarding the various aspects of learning the tests assess (stimulus pair learning, retrieval, and generalization of the previously learned associations to new but predictable stimulus pairs). In SP, behavioral performance was significantly poorer than in SF, and the reaction times were significantly longer, for all phases of the test. The results suggest that audiovisual associative learning is significantly influenced by the complexity of the consequent stimuli.

Vertical distribution of soil seed bank and the ecological importance of deeply buried seeds in alkaline grasslands.

Background: Soil seed banks play a central role in vegetation dynamics and may be an important source of ecological restoration. However, the vast majority of seed bank studies examined only the uppermost soil layers (0-10 cm); hence, our knowledge on the depth distribution of seed bank and the ecological significance of deeply buried seeds is limited. The aim of our study was to examine the fine-scale vertical distribution of soil seed bank to a depth of 80 cm, which is one of the largest studied depth gradients so far. Our model systems were alkaline grasslands in East-Hungary, characterised by harsh environmental conditions, due to Solonetz soil reference group with Vertic horizon. We asked the following questions: (1) How do the seedling density and species richness of soil seed bank change along a vertical gradient and to what depth can germinable seeds be detected? (2) What is the relationship between the depth distribution of the germinable seeds and the species traits? Methods: In each of the five study sites, four soil cores (4 cm diameter) of 80 cm depth were collected with an auger for soil seed bank analysis. Each sample was divided into sixteen 5-cm segments by depth (320 segments in total). Samples were concentrated by washing over sieves and then germinated in an unheated greenhouse. Soil penetration resistance was measured in situ next to each core location (0-80 cm depth, 1-cm resolution). We tested the number and species richness of seedlings observed in the soil segments (N = 320), using negative binomial generalized linear regression models, in which sampling layer and penetration resistance were the predictor variables. We ran the models for morphological groups (graminoids/forbs), ecological groups (grassland species/weeds) and life-form categories (short-lived/perennial). We also tested whether seed shape index, seed mass, water requirement or salt tolerance of the species influence the vertical distribution of their seed bank. Results: Germinable seed density and species richness in the seed bank decreased with increasing soil depth and penetration resistance. However, we detected nine germinable seeds of six species even in the deepest soil layer. Forbs, grassland species and short-lived species occurred in large abundance in deep layers, from where graminoids, weeds and perennial species were missing. Round-shaped seeds were more abundant in deeper soil layers compared to elongated ones, but seed mass and ecological indicator values did not influence the vertical seed bank distribution. Our research draws attention to the potential ecological importance of the deeply buried seeds that may be a source of recovery after severe disturbance. As Vertisols cover 335 million hectares worldwide, these findings can be relevant for many regions and ecosystems globally. We highlight the need for similar studies in other soil and habitat types to test whether the presence of deep buried seeds is specific to soils with Vertic characteristics.

The Influence of Stimulus Complexity on the Effectiveness of Visual Associative Learning.

Visually guided equivalence learning is a special type of associative learning, which can be evaluated using the Rutgers Acquired Equivalence Test (RAET) among other tests. RAET applies complex stimuli (faces and colored fish) between which the test subjects build associations. The complexity of these stimuli offers the test subject several clues that might ease association learning. To reduce the number of such clues, we developed an equivalence learning test (Polygon), which is structured as RAET but uses simple grayscale geometric shapes instead of faces and colored fish. In this study, we compared the psychophysical performances of the same healthy volunteers in both RAET and Polygon test. Equivalence learning, which is a basal ganglia-associated form of learning, appears to be strongly influenced by the complexity of the visual stimuli. The simple geometric shapes were associated with poor performance as compared to faces and fish. However, the difference in stimulus complexity did not affect performance in the retrieval and transfer parts of the test phase, which are assumed to be mediated by the hippocampi.

Investigation of Surface Properties and Free Volumes of Chitosan-Based Buccal Mucoadhesive Drug Delivery Films Containing Ascorbic Acid.

Nowadays, the buccal administration of mucoadhesive films is very promising. Our aim was to prepare ascorbic acid-containing chitosan films to study the properties and structures important for applicability and optimize the composition. During the formulation of mucoadhesive films, chitosan as the polymer basis of the film was used. Ascorbic acid, which provided the acidic pH, was used in different concentrations (2-5%). The films were formulated by the solvent casting method. The properties of films important for applicability were investigated, such as physical parameters, mucoadhesive force, surface free energy, and breaking strength. The fine structure of the films was analyzed by atomic force microscopy, and the free volume was analyzed by PALS, which can be important for drug release kinetics and the location of the drug in the film. The applicability of the optimized composition was also tested with two different types of active ingredients. The structure of the films was also analyzed by XRPD and FTIR. Ascorbic acid can be used well in chitosan films, where it can function as a permeation enhancer when reacting to chitosan, it is biodegradable, and can be applied in 2% of our studies.

Linking environmental heterogeneity and plant diversity: The ecological role of small natural features in homogeneous landscapes.

Small natural features (SNFs), such as road verges, midfield islets, rocky outcrops and ancient burial mounds, provide safe havens for species of natural habitats in human-modified landscapes; therefore, their great ecological importance is in contrast to their small size. SNFs often have a high topographical heterogeneity and abiotic conditions, which differ from their surroundings; therefore, they provide a unique opportunity for establishing links between environmental heterogeneity (EH) and biodiversity. However, no study has so far investigated the EH components of topographically heterogeneous SNFs in a comprehensive framework, by linking environmental and biotic parameters. To fill this knowledge gap, we evaluated the EH components and their effect on biodiversity on ancient mounds covered by semi-natural grasslands in the Pannonian (Hungary) and Continental (Bulgaria) biogeographical regions. We designated 16 study sites, each containing a few-metre-high mounds with five microsites (top, north-, east-, south- and west-facing slopes) and a nearby plain grassland. At each microsite, we measured soil moisture, soil chemical properties, solar radiation and microclimate; and recorded the cover of vascular plants in a total of 480 plots. On the mounds, topographical heterogeneity was associated with sharp differences in microclimate and soil properties. Besides the contrast between mild north-facing and harsh south-facing slopes, east- and west-facing slopes also sustained unique microsites characterised by dynamic diurnal changes in air temperature and vapour pressure deficit. Various combinations of the EH components resulted in unique plant species compositions within the microsites, and supported the co-occurrence of species typical of contrasting habitat types, even within a couple of metres. By combining high-resolution measurements of abiotic factors with fine-scale vegetation sampling, our study provides evidence that widespread SNFs with complex topography harbour several grassland-specialist plant species and introduce a high level of EH to otherwise homogeneous plain landscapes, which cover one third of the global land area.

Effects of Sucrose Palmitate on the Physico-Chemical and Mucoadhesive Properties of Buccal Films.

In our current research, sucrose palmitate (SP) was applied as a possible permeation enhancer for buccal use. This route of administration is a novelty as there is no literature on the use of SP in buccal mucoadhesive films. Films containing SP were prepared at different temperatures, with different concentrations of SP and different lengths of hydroxypropyl methylcellulose (HPMC) chains. The mechanical, structural, and in vitro mucoadhesive properties of films containing SP were investigated. Tensile strength and mucoadhesive force were measured with a device and software developed in our Institute. Positron annihilation lifetime spectroscopy (PALS) and X-ray powder diffractometry (XRPD) were applied for the structure analysis of the films. Mucoadhesive work was calculated in two ways: from the measured contact angle and compared with direct mucoadhesive work, which measured mucoadhesive force, which is direct mucoadhesion work. These results correlate linearly with a correlation coefficient of 0.98. It is also novel because it is a new method for the determination of mucoadhesive work.

Design and characterization of chitosan/citrate films as carrier for oral macromolecule delivery.

The oral delivery of biopharmaceuticals requires the including of absorption enhancer, protease inhibitor and a suitable carrier system. The aim of the present work was to formulate and characterize chitosan solutions/films incorporating citric acid (CA) as potential excipient in comparison to the well-known acetic acid (AA)-based films as a reference. Films were made by the solvent casting method with/without glycerol (G), propylene glycol (PG) and polyethylene glycol (PEG-400) as plasticizers. The minimum film forming temperature (MFFT) of the prepared solutions, film thickness, hardness/deformation, mucoadhesivity, moisture content, FT-IR spectra and surface free energy (SFE) were investigated. Chitosan has been reported as a safe and effective paracellular absorption enhancer for hydrophilic macromolecules, therefore there would be more rationale for incorporating CA as a solubility enhancer, a permeation enhancer and an enzyme inhibitor. CA shows good cross-linking, an ideal plasticizing property and increases both tensile strength and mucoadhesivity, thus its incorporation simplifies the formulation while improving effectiveness. We concluded that CA (3.5, 4 and 5 w/v %)-based chitosan solution could be used as a novel coating/subcoating polymer for oral macromolecule delivery, or as oral mucoadhesive films.

Preparation and investigation of core-shell nanoparticles containing human interferon-α.

Sustained release of active interferon-α (IFN-α) has been achieved from core-shell nanoparticles (NPs) prepared by aqueous precipitation of IFN-α-enriched human serum albumin (HSA-IFN-α) and layer-by-layer (L-b-L) by coating of the IFN-α NPs with poly(sodium-4-styrene) sulphonate (PSS) and chitosan (Chit). The concentration and the pH of HSA solution were optimized during the development of this method. Dynamic light scattering (DLS), zeta-potential, thermal analysis (differential scanning calorimetry (DSC) and termogravimetry (TG)), X-ray diffraction (XRD), IFN-α activity and morphology (transmission electron microscope (TEM)) studies were used to control the preparation and analyse the products. The dissolution kinetics of NPs was measured in vitro over 7 days in Hanson dissolution tester with Millex membrane. In vivo studies in Pannon white rabbit detected steady IFN-α plasma level for 10 days after subcutaneous injection administration of the HSA-IFN-α NPs. The IFN-α plasma concentration was detected by using the enzyme-linked immunosorbent assay (ELISA) method. In the present paper we discuss the preparation method, the optimization steps and the results of in vitro and in vivo release studies. It was established that 76.13% HSA-IFN-α are encapsulated in the core-shell NPs.

Density-Dependent Plant-Plant Interactions Triggered by Grazing.

Plant species performance in rangelands highly depends on the effect of grazing and also on the occurrence of unpalatable benefactor species that can act as biotic refuges protecting neighboring plants from herbivores. The balance between facilitation and competition may changes with the benefactor density. Despite the high number of studies on the role of biotic refuges, the density dependent effects of unpalatable herbaceous plants on the performance of other species, and on the habitat heterogeneity of rangelands are still unclear. Therefore, we performed a study to test the following hypotheses: (i) Performances of understory species follow a humped-back relationship along the density gradient of the unpalatable benefactor species. (ii) Small-scale heterogeneity of the vegetation decreases with increasing benefactor density. We studied meadow steppes with medium intensity cattle grazing in Hungary. We surveyed understory species' performance (number of flowering shoots and cover scores) along the density gradient of a common, native unpalatable species (Althaea officinalis). Our findings supported both hypotheses. We found unimodal relationship between the benefactor cover and both the flowering success and richness of understory species. Moreover, small-scale heterogeneity declined with increasing benefactor cover. In this study we detected a humped-back pattern of facilitation along the density gradient of an herbaceous benefactor in pastures. Indeed, this pattern was predictable based on such conceptual models like "consumer pressure-abiotic stress model," "humped-back model," "intermediate disturbance hypothesis," and "disturbance heterogeneity model"; but until now the validity of these relationships has not been demonstrated for herbaceous species. By the demonstration of this effect between herbaceous species we can better forecast the responses of grasslands to changes in management.

Both mass ratio effects and community diversity drive biomass production in a grassland experiment.

The maintenance of biodiversity is crucial for ecosystem processes such as plant biomass production, as higher species richness is associated with increased biomass production in plant communities. However, the effects of evenness and functional diversity on biomass production are understudied. We manipulated the composition of an experimental grassland by sowing various seed mixtures and examined the effects of diversity and evenness on biomass production after three years. We found that biomass production increased with greater species and functional richness but decreased with greater species and functional evenness. Standing biomass increased but species number and functional richness decreased with increasing proportion of perennial grasses. Our findings emphasise the importance of productive dominant species, as the proportion of perennial grasses had a positive effect on standing biomass, while species and functional evenness had a negative effect on it. Thus, our findings support the theory that, besides diversity, dominance effects and the so-called mass ratio hypothesis may also play a key role in explaining primary biomass production.

Vegetation type and grazing intensity jointly shape grazing effects on grassland biodiversity.

In the Palaearctic steppe zone, overgrazing was identified as one of the key drivers of declining grassland biodiversity, which underlines the necessity of the functional evaluation of increased grazing pressure on grassland vegetation. We tested the following hypotheses: (a) The effect of grazing intensity on species and functional diversity is strongly dependent on grassland type. (b) The magnitude of diet selectivity of grazers decreases with increasing grazing intensity. (c) Increasing grazing intensity increases evenness and functional evenness of the subjected grasslands. We analyzed vegetation patterns in four types of grasslands (Dry alkali short-grass steppes, Dry loess steppes, Non-alkali wet and Alkali wet grasslands) along an intensity gradient of beef cattle grazing at 73 sites in Hungary. Species richness, Shannon diversity, evenness, and four leaf traits were analyzed. We calculated community-weighted means for each single trait, and multi-trait functional richness, functional evenness, and divergence for all leaf traits. All species and functional diversity metrics were significantly affected by the grassland type, except leaf dry matter content. The effect of interaction between grazing intensity and grassland type was also significant for functional richness, functional evenness, community-weighted means of leaf area, and for species richness and evenness. An upward trend of specific leaf area was detected in all grasslands with the highest scores for the overgrazed sites, but the change was also grassland type dependent. The detected trend suggests that with increased intensity the overall selectivity of grazing decreased. We found that evenness was affected but functional evenness was not affected by grazing intensity. Functional evenness scores were more related to the grassland type than to changes in grazing intensity, and displayed a high variability. We stress that one-size-fits-all strategies cannot be recommended and actions should be fine-tuned at least at the level of grassland type.

Ecosystem engineering by foxes is mediated by the landscape context-A case study from steppic burial mounds.

In intensively used landscapes, remnant grassland fragments are often restricted to places unsuitable for agricultural cultivation. Such refuges are the ancient burial mounds called "kurgans," which are typical landscape elements of the Eurasian steppe and forest steppe zone. Due to their hill-like shape, loose soil structure and undisturbed status kurgans provide proper habitats for burrowing mammals. Accordingly, grassland vegetation on kurgans is often exposed to bioturbation, which can influence the habitat structure and plant species pool. In our study, we explored the effect of fox burrows and landscape context on the habitat properties and vegetation composition of small landscape elements, using kurgans as model habitats. We surveyed the vegetation of fox burrows and that of the surrounding grassland on five kurgans situated in cleared landscapes surrounded by arable lands and five kurgans in complex landscapes surrounded by grazed grasslands. We recorded the percentage cover of vascular plants, the amount of litter, and soil moisture content in twelve 0.5 m × 0.5 m plots per kurgan, in a total of 120 plots. We found that foxes considerably transformed habitat conditions and created microhabitats by changing the soil nutrient availability and reducing total vegetation cover and litter. Several grassland specialist species, mostly grasses (Agropyron cristatum, Elymus hispidus, and Stipa capillata) established in the newly created microhabitats, although the cover of noxious species was also considerable. We found that landscape context influenced the sort of species which could establish on kurgans by affecting the available species pool and soil moisture. Our results revealed that foxes act as ecosystem engineers on kurgans by transforming abiotic and biotic conditions by burrowing. Their engineering activity maintains disturbance-dependent components of dry grasslands and increases local environmental heterogeneity.

Litter removal does not compensate detrimental fire effects on biodiversity in regularly burned semi-natural grasslands.

Regulation of plant biomass accumulation is a key issue in effective grassland conservation in Europe. Burning is an alternative tool to regulate biomass dynamics in semi-natural grasslands even in the absence of grazing or mowing. We tested the effects of regular spring burning on the biomass fractions and fine-scale plant species composition of species-rich foothill grasslands in North-Hungary. There were five regularly burned and five control grasslands in the study; we collected twenty 20×20-cm sized biomass samples from each. We analyzed the main fractions (litter, graminoid and forb biomass), and the species-level biomass scores, and flowering success in the control and burned grasslands. We revealed that fire increased the amount of forb biomass and decreased the amount of litter, which suggested that regular burning might be feasible for regulating biomass dynamics. The non-metric multi-dimensional scaling (NMDS) showed a high similarity of the control and burned grasslands in species composition. However, plant diversity, and the number of flowering shoots decreased significantly in the burned grasslands. In regularly burned sites we found a significant decline of specialist species, as well as of steppic flora elements. Our results showed that besides its positive effect on biomass dynamics, high-frequency burning threatens the overall diversity and specialist plant species in semi-natural grasslands. We recommend that proper fire regimes should be first studied experimentally, to provide a scientific basis for the application of prescribed burning management in such habitats.

New aspects of grassland recovery in old-fields revealed by trait-based analyses of perennial-crop-mediated succession.

Classical old-field succession studies focused on vegetation changes after the abandonment of annual croplands or on succession after the elimination of cultivated crops. Perennial-crop-mediated succession, where fields are initially covered by perennial crops, reveals alternative aspects of old-field succession theories. We tested the validity of classical theories of old-field succession for perennial-crop-mediated succession. We formulated the following hypotheses: (1) functional diversity increases with increasing field age; (2) resource acquisition versus conservation trade-off shifts toward conservation at community level during the succession; (3) the importance of spatial and temporal seed dispersal decreases during the succession; and (4) competitiveness and stress-tolerance increases and ruderality decreases at community level during the succession. We studied functional diversity, trait distributions and plant strategies in differently aged old-fields using chronosequence method. We found increasing functional richness and functional divergence, but also unchanged or decreasing functional evenness. We detected a shift from resource acquisition to resource conservation strategy of communities during the succession. The role of spatial and temporal seed dispersal was found to be important not only at the initial but also at latter successional stages. We found an increasing stress-tolerance and a decreasing ruderality during succession, while the competitiveness remained unchanged at the community level. Despite the markedly different starting conditions, we found that classical and perennial-crop-mediated old-field successions have some similarities regarding the changes of functional diversity, resource acquisition versus conservation trade-off, and seed dispersal strategies. However, we revealed also the subsequent differences. The competitive character of communities remained stable during the succession; hence, the initial stages of perennial-crop-mediated succession can be similar to the middle stages of classical old-field succession. Moreover, the occupied functional niche space and differentiation were larger in the older stages, but resources were not effectively utilized within this space, suggesting that the stabilization of the vegetation requires more time.