Biochemical composition changes can be linked to the tolerance of four grassland species under more persistent precipitation regimes.

Climate models suggest that the persistence of summer precipitation regimes (PRs) is on the rise, characterized by both longer dry and longer wet durations. These PR changes may alter plant biochemical composition and thereby their economic and ecological characteristics. However, impacts of PR persistence have primarily been studied at the community level, largely ignoring the biochemistry of individual species. Here, we analyzed biochemical components of four grassland species with varying sensitivity to PR persistence (Holcus lanatus, Phleum pratense, Lychnis flos-cuculi, Plantago lanceolata) along a range of increasingly persistent PRs (longer consecutive dry and wet periods) in a mesocosm experiment. The more persistent PRs decreased nonstructural sugars, whereas they increased lignin in all species, possibly reducing plant quality. The most sensitive species Lychnis seemed less capable of altering its biochemical composition in response to altered PRs, which may partly explain its higher sensitivity. The more tolerant species may have a more robust and dynamic biochemical network, which buffers the effects of changes in individual biochemical components on biomass. We conclude that the biochemical composition changes are important determinants for plant performance under increasingly persistent precipitation regimes.

Community evolution increases plant productivity at low diversity.

Species extinctions from local communities negatively affect ecosystem functioning. Ecological mechanisms underlying these impacts are well studied, but the role of evolutionary processes is rarely assessed. Using a long-term field experiment, we tested whether natural selection in plant communities increased biodiversity effects on productivity. We re-assembled communities with 8-year co-selection history adjacent to communities with identical species composition but no history of co-selection ('naïve communities'). Monocultures, and in particular mixtures of two to four co-selected species, were more productive than their corresponding naïve communities over 4 years in soils with or without co-selected microbial communities. At the highest diversity level of eight plant species, no such differences were observed. Our findings suggest that plant community evolution can lead to rapid increases in ecosystem functioning at low diversity but may take longer at high diversity. This effect was not modified by treatments simulating co-evolutionary processes between plants and soil organisms.

Elevated CO2 and water addition enhance nitrogen turnover in grassland plants with implications for temporal stability.

Temporal variation in soil nitrogen (N) availability affects growth of grassland communities that differ in their use and reuse of N. In a 7-year-long climate change experiment in a semi-arid grassland, the temporal stability of plant biomass production varied with plant N turnover (reliance on externally acquired N relative to internally recycled N). Species with high N turnover were less stable in time compared to species with low N turnover. In contrast, N turnover at the community level was positively associated with asynchrony in biomass production, which in turn increased community temporal stability. Elevated CO2 and summer irrigation, but not warming, enhanced community N turnover and stability, possibly because treatments promoted greater abundance of species with high N turnover. Our study highlights the importance of plant N turnover for determining the temporal stability of individual species and plant communities affected by climate change.

Root traits are more than analogues of leaf traits: the case for diaspore mass.

Root traits are often thought to be analogues of leaf traits along the plant economics spectrum. But evolutionary pressures have most likely shaped above- and belowground patterns differentially. Here, we aimed to identify the most important aboveground traits for explaining root traits without an a priori focus on known concepts. We measured morphological root traits in a glasshouse experiment on 141 common Central European grassland species. Using random forest algorithms, we built predictive models of six root traits from 97 aboveground morphological, ecological and life history traits. Root tissue density was best predicted by leaf dry matter content, whereas traits related to root fineness were best predicted by diaspore mass: the heavier the diaspore, the coarser the root system. Specific leaf area (SLA) was not an important predictor for any of the root traits. This study confirms the hypothesis that root traits are more than analogues of leaf traits within a plant economics spectrum. The results reveal a novel ecological pattern and highlight the power of root data to close important knowledge gaps in trait-based ecology.

Dry matter yield, chemical composition and estimated extractable protein of legume and grass species during the spring growth.

BACKGROUND: Knowledge of the variation of extractable protein amount in legumes and grasses as affected by harvest time is important for identifying optimal combinations to enable a high protein production in a biorefinery. The extractability of protein was estimated using the Cornell Net Carbohydrate and Protein System across six harvests during the spring growth. RESULTS: The estimated extractable protein [g kg-1 dry matter (DM)] defined as the easily available fractions B1 +B2 was significantly higher in white clover and lucerne at all harvests while, if the more cell wall attached fraction B3 can be extracted, white clover had the highest extractable protein amongst all species. Total yield of B1 +B2 per ha was higher in white clover and red clover at the early growth while B1 +B2 +B3 was by far the highest for red clover through all harvests. CONCLUSION: White clover could be a good candidate for protein production purpose in a biorefinery due to its high extractable protein content per kg DM. In order to maximise the protein production capacity, harvest should take place during early growth due to a decline in protein extractability with maturity. The final economy of the concept will depend on the value of the fibre after extraction of the protein. © 2017 Society of Chemical Industry.

Grassland species differentially regulate proline concentrations under future climate conditions: an integrated biochemical and modelling approach.

Proline (Pro) is a versatile metabolite playing a role in the protection of plants against environmental stresses. To gain a deeper understanding of the regulation of Pro metabolism under predicted future climate conditions, including drought stress, elevated temperature and CO2 , we combined measurements in contrasting grassland species (two grasses and two legumes) at multiple organisational levels, that is, metabolite concentrations, enzyme activities and gene expression. Drought stress (D) activates Pro biosynthesis and represses its catabolism, and elevated temperature (DT) further elevated its content. Elevated CO2 attenuated the DT effect on Pro accumulation. Computational pathway control analysis allowed a mechanistic understanding of the regulatory changes in Pro metabolism. This analysis indicates that the experimentally observed coregulation of multiple enzymes is more effective in modulating Pro concentrations than regulation of a single step. Pyrroline-5-carboxylate synthetase (P5CS) and pyrroline-5-carboxylate reductase (P5CR) play a central role in grasses (Lolium perenne, Poa pratensis), and arginase (ARG), ornithine aminotransferase (OAT) and P5CR play a central role in legumes (Medicago lupulina, Lotus corniculatus). Different strategies in the regulation of Pro concentrations under stress conditions were observed. In grasses the glutamate pathway is activated predominantly, and in the legumes the ornithine pathway, possibly related to differences in N-nutritional status.

Seed dispersal limitation causes negative legacy effect on restoration of grassland plant diversity on ski slopes.

Past forest use often has a long-term negative impact on the recovery of the original plant composition of semi-natural grasslands, which is known as a legacy effect. This study investigates the impact of seed dispersal limitations on the restoration of grassland plant diversity on ski slopes with past forest use, highlighting the negative legacy effect on biodiversity recovery. Focusing on ski areas, our research contrasts the vegetation on ski slopes originally created on semi-natural grasslands such as pasture (pasture slopes) and constructed by clearing secondary forests or conifer plantations (forest slopes). We examined species richness and diversity, considering seed dispersal modes, grassland management history, and seed source proximity. We reveal that the proximity to species-rich grassland sources is pivotal for the restoration of native grassland vegetation. Particularly, wind-dispersed species show significant recovery on slopes with sustained management for more than 70 years and those with neighboring species-rich grasslands, suggesting that both the duration of management and the proximity to seed sources are critical for overcoming the legacy effects of past forest use. Meanwhile, gravity-dispersed species failed to recover their richness and diversity regardless of both the duration of management and the proximity to seed source grasslands, which their diversity recovered where seed sources neighbored. Our findings emphasize the importance of considering seed dispersal limitation and management history in the restoration and conservation of grasslands and their biodiversity, particularly in landscapes experiencing past human intervention.

Drought induces moderate, diverse changes in the odour of grassland species.

Plants react to drought stress with numerous changes including altered emissions of volatile organic compounds (VOC) from leaves, which provide protection against oxidative tissue damage and mediate numerous biotic interactions. Despite the share of grasslands in the terrestrial biosphere, their importance as carbon sinks and their contribution to global biodiversity, little is known about the influence of drought on VOC profiles of grassland species. Using coupled gas chromatography-mass spectrometry, we analysed the odorants emitted by 22 European grassland species exposed to an eight-week-lasting drought treatment (DT; 30% water holding capacity, WHC). We focused on the odorants emitted during the light phase from whole plant shoots in their vegetative stage. Emission rates were standardised to the dry weight of each shoot. Well-watered (WW) plants (70% WHC) served as control. Drought-induced significant changes included an increase in total emission rates of plant VOC in six and a decrease in three species. Diverging effects on the number of emitted VOC (chemical richness) or on the Shannon diversity of the VOC profiles were detected in 13 species. Biosynthetic pathways-targeted analyses revealed 13 species showing drought-induced higher emission rates of VOC from one, two, three, or four major biosynthetic pathways (lipoxygenase, shikimate, mevalonate and methylerythritol phosphate pathway), while six species exhibited reduced emission rates from one or two of these pathways. Similarity trees of odorant profiles and their drought-induced changes based on a biosynthetically informed distance metric did not match species phylogeny. However, a phylogenetic signal was detected for the amount of terpenoids released by the studied species under WW and DT conditions. A comparative analysis of emission rates of single compounds released by WW and DT plants revealed significant VOC profile dissimilarities in four species only. The moderate drought-induced changes in the odorant emissions of grassland species are discussed with respect to their impact on trophic interactions across the food web. (294 words).

Inoculation with Arbuscular Mycorrhizal Fungi Supports the Uptake of Macronutrients and Promotes the Growth of Festuca ovina L. and Trifolium medium L., a Candidate Species for Green Urban Infrastructure.

Green roofs and walls play an important role in promoting biodiversity, reducing the urban heat island effect and providing ecosystem services in urban areas. However, the conditions on green walls/roofs (low nutrient and organic matter content, drought, high temperatures) are often unfavorable for plant growth. Arbuscular mycorrhizal fungi (AMF) can improve the growth and development of plants under stress conditions as they can increase nutrient and water uptake. In a 6-month pot experiment, we investigated the effect of AMF inoculation on the growth and NPK uptake of Festuca ovina L. and Trifolium medium L., which are used for green roofs and walls. Two variants of mycorrhizal inoculation were used in the experiment: a commercial mycorrhizal inoculant AM Symbivit (Symbiom Ltd., Lanskroun, Czech Republic) and a mycorrhizal inoculant collected from calcareous grassland in the Silesia region (Poland). Funneliformis mosseae was the most abundant species in the roots of F. ovina and T. medium with IM inoculum. In the CM variant, a dominance of F. mosseae was observed in the roots of F. ovina. In contrast, Archaeosporaceae sp. node 317 dominated in the roots of T. medium. Both inoculations had a positive effect on the increase in dry weight of the shoots of T. medium, but only the commercial inoculum had a positive effect on the growth of F. ovina. Both inoculations improved the P uptake by the roots and the P and K uptake into the shoots of T. medium. In addition, both inoculations improved the K uptake by the roots of F. ovina and the N, P and K uptake into the shoots. In conclusion, both AMF communities included in the inoculations had a positive effect on plant growth and nutrient uptake, but the effect depends on the plant and the mycorrhizal fungus species.

The Effect of Combined Drought and Temperature Stress on the Physiological Status of Calcareous Grassland Species as Potential Candidates for Urban Green Infrastructure.

Nature-based solutions are promising for climate adaptation and environmental management in urban areas, but urban conditions are stressful for vegetation. In particular, the interaction of drought and high temperatures may be detrimental. Guiding plant selection for urban greening with native species requires a far better knowledge of plant adaptations and stress acclimation. We tested the physiological responses of four candidate calcareous grassland species for green roofs and walls to the combined effects of drought and high temperatures under controlled conditions. The tested species proved relatively resistant to stress despite different strategies to protect the photosynthetic apparatus, maintain water balance, and repair damages. Based on the physiological responses, we rank the species in descending order of resistance to the stress factors tested: Trifolium medium > Festuca ovina > Carex flacca > Potentilla reptans, but all four can serve as potential candidates for green walls and roofs. Physiological stress screening of plant species for use on green roofs and walls supplements the habitat template approach to provide a stronger and wider base for prioritizations.

Does previous exposure to extreme precipitation regimes result in acclimated grassland communities?

Climate change will likely increase weather persistence in the mid-latitudes, resulting in precipitation regimes (PR) with longer dry and wet periods compared to historic averages. This could affect terrestrial ecosystems substantially through the increased occurrence of repeated, prolonged drought and water logging conditions. Climate history is an important determinant of ecosystem responses to consecutive environmental extremes, through direct damage, community restructuring as well as morphological and physiological acclimation in species or individuals. However, it is unclear how community restructuring and individual metabolic acclimation effects interact to determine ecosystem responses to subsequent climate extremes. Here, we investigated, if and how, differences in exposure to extreme or historically normal PR induced long-lasting (i.e. legacy) effects at the level of community (e.g., species composition), plant (e.g., biomass), and molecular composition (e.g., sugars, lipids, stress markers). Experimental grassland communities were exposed to long (extreme) or short (historically normal) dry/wet cycles in year 1 (Y1), followed by exposure to an identical PR or the opposite PR in year 2 (Y2). Results indicate that exposure to extreme PR in Y1, reduced diversity but induced apparent acclimation effects in all climate scenarios, stimulating biomass (higher productivity and structural sugar content) in Y2. In contrast, plants pre-exposed to normal PR, showed more activated stress responses (higher proline and antioxidants) under extreme PR in Y2. Overall, Y1 acclimation effects were strongest in the dominant grasses, indicating comparatively high phenotypical plasticity. However, Y2 drought intensity also correlated with grass productivity and structural sugar findings, suggesting that responses to short-term soil water deficits contributed to the observed patterns. Interactions between different legacy effects are discussed. We conclude that more extreme PR will likely alter diversity in the short-to midterm and select for acclimated grassland communities with increased productivity and attenuated molecular stress responses under future climate regimes.

Silicon Effects on Biomass Carbon and Phytolith-Occluded Carbon in Grasslands Under High-Salinity Conditions.

Changes in climate and land use are causing grasslands to suffer increasingly from abiotic stresses, including soil salinization. Silicon (Si) amendment has been frequently proposed to improve plant resistance to multiple biotic and abiotic stresses and increase ecosystem productivity while controlling the biogeochemical carbon (C) cycle. However, the effects of Si on plant C distribution and accumulation in salt-suffering grasslands are still unclear. In this study, we investigated how salt ions affected major elemental composition in plants and whether Si enhanced biomass C accumulation in grassland species in situ. In samples from the margins of salt lakes, our results showed that the differing distance away from the shore resulted in distinctive phytocoenosis, including halophytes and moderately salt-tolerant grasses, which are closely related to changing soil properties. Different salinity (Na+/K+, ranging from 0.02 to 11.8) in plants caused negative effects on plant C content that decreased from 53.9 to 29.2% with the increase in salinity. Plant Si storage [0.02-2.29 g Si m-2 dry weight (dw)] and plant Si content (0.53 to 2.58%) were positively correlated with bioavailable Si in soils (ranging from 94.4 to 192 mg kg-1). Although C contents in plants and phytoliths were negatively correlated with plant Si content, biomass C accumulation (1.90-83.5 g C m-2 dw) increased due to the increase of Si storage in plants. Plant phytolith-occluded carbon (PhytOC) increased from 0.07 to 0.28‰ of dry mass with the increase of Si content in moderately salt-tolerant grasses. This study demonstrates the potential of Si in mediating plant salinity and C assimilation, providing a reference for potential manipulation of long-term C sequestration via PhytOC production and biomass C accumulation in Si-accumulator dominated grasslands.

Variable Influences of Water Availability and Rhizobacteria on the Growth of Schizachyrium scoparium (Little Bluestem) at Different Ages.

There is significant interest in understanding the role of plant growth-promoting rhizobacteria (PGPR) in alleviating different types of plant stress. Schizachyrium scoparium (little bluestem) is a moderately drought tolerant, perennial bunchgrass native to North America. The goal of this experiment was to evaluate whether the addition of a bacterial root isolate in the Pseudomonas genus promoted the growth of S. scoparium with changes in water availability. Pseudomonas are common rhizobacteria and have been shown to improve plant growth. It was hypothesized that plants inoculated with the PGPR strain would have greater growth and health, and would be less affected by shifts in water availability. Pseudomonas strains were isolated from the roots of native S. scoparium plants. After germination, S. scoparium seedlings were subjected to four treatment groups: low water; high water; low water with PGPR; and high water with PGPR. The experiment was run three times with plants at different starting ages; 14-, 28-, and 70-day-old plants. The effects of the water and PGPR treatments were variable between the experimental trials. There were no significant effects of the water treatments on plant growth in Trial 1 (14-day-old plants) or Trial 2 (28-day-old plants), however, there was a significant negative effect of the high watering treatment on the shoot length and biomass in Trial 3. High water availability was significantly associated with greater plant health in Trial 1, but appeared to reduce plant health in Trials 2 and 3. The PGPR treatment appeared to promote root growth and biomass in Trial 2, and was associated with greater plant health in all three trials, especially when paired with the low water treatment. Results from a permutational MANOVA indicate that plant growth was significantly different between the trials due to differences in the starting age of the plants and the duration of the experiments. Thus, methodological choices, such as plant life history stage and experiment duration, may affect the response of plants to PGPR in the rhizosphere. This research provides an insight into the interactions between PGPR and water availability on the growth and health of native plants.

Predicting methane yield by linear regression models: A validation study for grassland biomass.

The objectives of this study were to assess and validate previously published prediction models with an independent dataset and to expose the power and limitations of linear regression models for predicting biomethane potential. Two datasets were used for the validation, one with all individual samples and one with the average values of each cultivar. The results revealed similar performances of all four models for the individual samples. The methane yields of the cultivars were predicted more accurately than the methane yields of the individual samples. The grassland specific model predicted the variation in the dataset with an R2 of 0.84 and the slope of the regression line was equal to 1.0. Linear regression models are suitable to depict the variation in methane yield and for substrate ranking. However, the prediction error of the absolute values may be high since systematic external effects cannot be determined by a regression model.

Redescription of Yphthimoides patricia (Hayward, 1957), with taxonomic notes on the names Euptychia saltuensis Hayward, 1962 and Yphthimoides manasses (C. Felder R. Felder, 1867) (Nymphalidae: Satyrinae).

Euptychia saltuensis Hayward, 1962, new synonym, currently regarded as a nomen dubium and possibly a junior subjective synonym of Yphthimoides manasses (C. Felder R. Felder, 1867), is here treated as a junior subjective synonym of Yphthimoides patricia (Hayward, 1957), based on morphological characters of the male genitalia and the DNA barcode. The taxonomic status of Y. patricia is re-examined, and a detailed redescription of the adult morphology, including the male genitalia, is presented. Information on the distribution, habitat and immature stages of Y. patricia is also provided. Yphthimoides patricia is clearly a distinct species from Y. manasses based on the analysis of DNA barcode sequences and the morphology of the male genitalia.

Reduced germination success of temperate grassland seeds sown in dung: consequences for post-dispersal seed fate.

Endozoochory is one of the main drivers shaping temperate grassland communities by maintaining plant populations of its constituents and enabling plants to colonize new habitats. Successful endozoochorous dispersal implies that seeds not only get consumed and survive the digestive tract but are also able to develop into viable seedlings in a dung environment. We experimentally assessed the germination probability and timing of 15 annual and perennial temperate European grassland species in cattle and horse dung and in different climatic conditions (greenhouse and outdoor conditions). Interspecific variation in germinability and germination timing are found, while life strategy had only an effect on germination timing. We found adverse effects of both cattle and horse dung on the germination characteristics of all tested grassland species, but the effects of cattle dung were more pronounced. In comparison with the control treatment, fewer seeds emerged in dung and more time was needed to germinate. Also, germination metrics clearly differed between the artificial greenhouse and outdoor conditions, with generally a lower germinability in outdoor conditions. According to our results, a large cost seems to be associated with endozoochorous dispersal in this stage of the life cycle, as seed dispersal effectiveness strongly depends on the quality of the deposition site with a lowered survival and germination probability when seeds are deposited in dung.

Elevated CO2 mitigates drought and temperature-induced oxidative stress differently in grasses and legumes.

Increasing atmospheric CO2 will affect plant growth, including mitigation of stress impact. Such effects vary considerably between species-groups. Grasses (Lolium perenne, Poa pratensis) and legumes (Medicago lupulina, Lotus corniculatus) were subjected to drought, elevated temperature and elevated CO2. Drought inhibited plant growth, photosynthesis and stomatal conductance, and induced osmolytes and antioxidants in all species. In contrast, oxidative damage was more strongly induced in the legumes than in the grasses. Warming generally exacerbated drought effects, whereas elevated CO2 reduced stress impact. In the grasses, photosynthesis and chlorophyll levels were more protected by CO2 than in the legumes. Oxidative stress parameters (lipid peroxidation, H2O2 levels), on the other hand, were generally more reduced in the legumes. This is consistent with changes in molecular antioxidants, which were reduced by elevated CO2 in the grasses, but not in the legumes. Antioxidant enzymes decreased similarly in both species-groups. The ascorbate-glutathione cycle was little affected by drought and CO2. Overall, elevated CO2 reduced drought effects in grasses and legumes, and this mitigation was stronger in the legumes. This is possibly explained by stronger reduction in H2O2 generation (photorespiration and NADPH oxidase), and a higher availability of molecular antioxidants. The grass/legume-specificity was supported by principal component analysis.

Discrimination of grassland species and their classification in botanical families by laboratory scale NIR hyperspectral imaging: preliminary results.

The objective of this study was to discriminate by a NIR line scan hyperspectral imaging, taxonomic plant families comprised of different grassland species. Plants were collected from semi-natural meadows of the National Apuseni Park, Apuseni Mountains, Gârda area (Romania) according to botanical families. Chemometric tools such as PLS-DA were used to discriminate distinct grassland species, and assign the different species to botanical families. Species within the Poacea family and other Botanical families could be distinguished (R(2)=0.91 and 0.90, respectively) with greater accuracy than those species in the Fabacea family (R(2)=0.60). A correct classification rate of 99% was obtained in the assignment of the various species to the proper family. Moreover a complete study based on wavelength selection has been performed in order to identify the chemical compound related to each botanical family and therefore to the possible toxicity of the plant. This work could be considered as a first step for the development of a complete procedure for the detection and quantification of possible toxic species in semi-natural meadows used by grazing animals.

A simple method for in situ-labelling with 15N and 13C of grassland plant species by foliar brushing.

1.Labelling plants with 15N and 13C stable isotopes usually require cultivation of plants in isotopically enriched soil and gas-tight labelling chambers - both approaches are not suitable if one aims to investigate in situ species interactions in real plant communities. 2.In this greenhouse experiment, we tested a labelling method in which dual-labelled (15N, 13C) urea solution is brushed directly onto leaves of twelve temperate grassland species representing grasses, non-leguminous forbs and legumes. 3.Across all plant species, shoots (15N: 0·145; 13C: 0·090 atom percent excess, APE) and roots (15N: 0·051; 13C: 0·023 APE) were significantly enriched after five daily labelling events. Generally, isotopic enrichments were significantly higher in shoots than in roots. No clear pattern of absolute isotopic enrichment was observed between plant functional groups; however, grasses showed a more even allocation between shoots and roots than forbs and legumes. Isotopic enrichment levels after 4 weeks were lower, higher or unchanged compared to those of week one and varied between species or plant parts. 4.Considering the consistent enrichment levels and simplicity of this method, we conclude that it can be applied widely in ecological studies of above-belowground plant-plant or plant-animal interactions even in real plant communities.