PHYLOGENETIC ANALYSES, GENE DOSAGE EFFECT AND SELECTION IN MEMBERS OF THE LATERAL BOUNDARY DOMAIN (LBD) GENES TRANSCRIPTION FACTOR FAMILY

Wood formation is an economically and environmentally important process and has played a significant role in the evolution of terrestrial plants. Despite its significance, the molecular underpinnings of the process are still poorly understood. We have previously shown that four Lateral Boundary Domain (LBD) transcription factors have important roles in the regulation of wood formation with two (LBD1 and LBD4) involved in secondary phloem and ray cell development and two (LBD15 and LBD18) in secondary xylem formation. Here, we used comparative phylogenetic analyses to test potential roles of the four LBD genes in the evolution of woodiness. We studied the copy number and variation in DNA and amino acid sequences of the four LBDs in a wide range of woody and herbaceous plant taxa with fully sequenced and annotated genomes. LBD1 showed the highest gene copy number across the studied species, and LBD1 gene copy number was strongly and significantly correlated with the level of ray seriation. The lianas, cucumber and grape, with multiseriate ray cells showed the highest gene copy number (12 and 11, respectively). Because lianas’ growth habit requires significant twisting and bending, the less lignified ray parenchyma cells likely facilitate stem flexibility and maintenance of xylem conductivity. We further demonstrate conservation of amino acids in the LBD18 protein sequences that are specific to woody taxa. Neutrality tests showed evidence for strong purifying selection on these gene regions across various orders, indicating adaptive convergent evolution of LBD18. Structural modeling demonstrates that the conserved amino acids have a significant impact on the tertiary protein structure and thus are likely of significant functional importance.

The Effect of Hydrogen Overvoltage on the Electrolytic Deposition of Zinc

In the deposition of metallic zinc by electro­lysis from neutral or acid solution, little difficulty is experienced provided certain impurities are absent from the electrolyte. The use of the process has long been considered as a potential source, patents on the process having been issued as early as 1880. However, the early experimenters failed to realize the importance of impurities in the electrolyte, and for this reason, the process suffered several severe setbacks when commercial plants were built.

Homeopathic treatment of Arabidopsis thaliana plants infected with Pseudomonas syringae

Homeopathic basic research is still in the screening phase to identify promising model systems that are adapted to the needs and peculiarities of homeopathic medicine and pharmacy. We investigated the potential of a common plant-pathogen system, Arabidopsis thaliana infected with the virulent bacteria Pseudomonas syringae, regarding its response towards a homeopathic treatment. A. thaliana plants were treated with homeopathic preparations before and after infection. Outcome measure was the number of P. syringae bacteria in the leaves of A. thaliana, assessed in randomized and blinded experiments. After a screening of 30 homeopathic preparations, we investigated the effect of Carbo vegetabilis 30x, Magnesium phosphoricum 30x, Nosode 30x, Biplantol (a homeopathic complex remedy), and Biplantol 30x on the infection rate in five or six independent experiments in total. The screening yielded significant effects for four out of 30 tested preparations. In the repeated experimental series, only the homeopathic complex remedy Biplantol induced a significant reduction of the infection rate (p = 0.01; effect size, d = 0.38). None of the other four repeatedly tested preparations (Carbo vegetabilis 30x, Magnesium phosphoricum 30x, Nosode 30x, Biplantol 30x) yielded significant effects in the overall evaluation. This phytopathological model yielded a small to medium effect size and thus might be of interest for homeopathic basic research after further improvement. Compared to Bion (a common SAR inducer used as positive control), the magnitude of the treatment effect of Biplantol was about 50%. Thus, homeopathic formulations might have a potential for the treatment of plant diseases after further optimization. However, the ecological impact should be investigated more closely before widespread application.

Different Land Use Intensities in Grassland Ecosystems Drive Ecology of Microbial Communities Involved in Nitrogen Turnover in Soil

Understanding factors driving the ecology of N cycling microbial communities is of central importance for sustainable land use. In this study we report changes of abundance of denitrifiers, nitrifiers and nitrogen-fixing microorganisms (based on qPCR data for selected functional genes) in response to different land use intensity levels and the consequences for potential turnover rates. We investigated selected grassland sites being comparable with respect to soil type and climatic conditions, which have been continuously treated for many years as intensely used meadows (IM), intensely used mown pastures (IP) and extensively used pastures (EP), respectively. The obtained data were linked to above ground biodiversity pattern as well as water extractable fractions of nitrogen and carbon in soil. Shifts in land use intensity changed plant community composition from systems dominated by s-strategists in extensive managed grasslands to c-strategist dominated communities in intensive managed grasslands. Along the different types of land use intensity, the availability of inorganic nitrogen regulated the abundance of bacterial and archaeal ammonia oxidizers. In contrast, the amount of dissolved organic nitrogen determined the abundance of denitrifiers (nirS and nirK). The high abundance of nifH carrying bacteria at intensive managed sites gave evidence that the amounts of substrates as energy source outcompete the high availability of inorganic nitrogen in these sites. Overall, we revealed that abundance and function of microorganisms involved in key processes of inorganic N cycling (nitrification, denitrification and N fixation) might be independently regulated by different abiotic and biotic factors in response to land use intensity.

Biodiversity Effects on Plant Stoichiometry

In the course of the biodiversity-ecosystem functioning debate, the issue of multifunctionality of species communities has recently become a major focus. Elemental stoichiometry is related to a variety of processes reflecting multiple plant responses to the biotic and abiotic environment. It can thus be expected that the diversity of a plant assemblage alters community level plant tissue chemistry. We explored elemental stoichiometry in aboveground plant tissue (ratios of carbon, nitrogen, phosphorus, and potassium) and its relationship to plant diversity in a 5-year study in a large grassland biodiversity experiment (Jena Experiment). Species richness and functional group richness affected community stoichiometry, especially by increasing C:P and N:P ratios. The primacy of either species or functional group richness effects depended on the sequence of testing these terms, indicating that both aspects of richness were congruent and complementary to expected strong effects of legume presence and grass presence on plant chemical composition. Legumes and grasses had antagonistic effects on C:N (−27.7% in the presence of legumes, +32.7% in the presence of grasses). In addition to diversity effects on mean ratios, higher species richness consistently decreased the variance of chemical composition for all elemental ratios. The diversity effects on plant stoichiometry has several non-exclusive explanations: The reduction in variance can reflect a statistical averaging effect of species with different chemical composition or a optimization of nutrient uptake at high diversity, leading to converging ratios at high diversity. The shifts in mean ratios potentially reflect higher allocation to stem tissue as plants grew taller at higher richness. By showing a first link between plant diversity and stoichiometry in a multiyear experiment, our results indicate that losing plant species from grassland ecosystems will lead to less reliable chemical composition of forage for herbivorous consumers and belowground litter input.

Holocene climatic change and the development of the lake-effect snowbelt in Michigan, USA

Lake-effect snow is an important constraint on ecological and socio-economic systems near the North American Great Lakes. Little is known about the Holocene history of lake-effect snowbelts, and it is difficult to decipher how lake-effect snowfall abundance affected ecosystem development. We conducted oxygen-isotope analysis of calcite in lake-sediment cores from northern Lower Michigan to infer Holocene climatic variation and assess snowbelt development. The two lakes experience the same synoptic-scale climatic systems, but only one of them (Huffman Lake) receives a significant amount of lake-effect snow. A 177-cm difference in annual snowfall causes groundwater inflow at Huffman Lake to be 18O-depleted by 2.3‰ relative to O'Brien Lake. To assess when the lake-effect snowbelt became established, we compared calcite-δ18O profiles of the last 11,500 years from these two sites. The chronologies are based on accelerator-mass-spectrometry 14C ages of 11 and 17 terrestrial-plant samples from Huffman and O'Brien lakes, respectively. The values of δ18O are low at both sites from 11,500 to 9500 cal yr BP when the Laurentide Ice Sheet (LIS) exerted a dominant control over the regional climate and provided periodic pulses of meltwater to the Great Lakes basin. Carbonate δ18O increases by 2.6‰ at O'Brien Lake and by 1.4‰ at Huffman Lake between 9500 and 7000 cal yr BP, suggesting a regional decline in the proportion of runoff derived from winter precipitation. The Great Lakes snowbelt probably developed between 9500 and 5500 cal yr BP as inferred from the progressive 18O-depletion at Huffman Lake relative to O'Brien Lake, with the largest increase of lake-effect snow around 7000 cal yr BP. Lake-effect snow became possible at this time because of increasing contact between the Great Lakes and frigid arctic air. These changes resulted from enhanced westerly flow over the Great Lakes as the LIS collapsed, and from rapidly rising Great Lakes levels during the Nipissing Transgression. The δ18O difference between Huffman and O'Brien lakes declines after 5500 cal yr BP, probably because of a northward shift of the polar vortex that brought increasing winter precipitation to the entire region. However, δ18O remains depleted at Huffman Lake relative to O'Brien Lake because of the continued production of lake-effect snow.

Differences between recent and historical records of upperspecies limits in the northern European Alps

Are there differences in historical and recent upper range limits of vascular plants and are such differences more pronounced in individual species groups? The limits of 1103 plants of the Northern Alps are compared to range limits in the mid-19th century. The comparison is based on two surveys. The first survey was conducted by Otto Sendtner in 1848–1853, the second in 1991–2008 during a habitat inventory. To our knowledge this is the first comparative studies reaching back to the end of the “Little Ice Age” and comprising an almost entire regional flora covering the complete range of habitats. During the recent survey, most species were found at higher elevations. Even though the differences fit well with the expected shifts due to climate warming we cannot exclude effects of sampling bias. However, we assume that the relative differences between species groups can be safely interpreted. The differences in upper limits between both surveys were significantly larger among forest species. The most important reason is probably discontinued pasture and mowing, which may have amplified possible warming effects. Nitrogen deposits may have contributed to this effect by placing competitive species in a more advantageous position.

The Effect of Heat Treatment of Forages on Degradation Kinetics and Escape Protein Concentration

An in situ study was conducted to evaluate the effects of heat treatments on the degradation kinetics and escape protein concentrations of forages (alfalfa and berseem clover). Alfalfa collected at 4 and 7 weeks post-harvest and berseem clover collected at 5 and 7 weeks postharvest were freeze-dried and then heated to 100, 125, and 150o C for 2 hours. Heat treatment effects were determined by placing two bags of sample (for each treatment, maturity, and forage species for a given incubation times) into the rumen of one fistulated steer fed alfalfa hay. Bags were incubated for periods of 0 to 48 hours. Increasing levels of heat treatments of forages increased concentrations of neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent insoluble nitrogen (ADIN) and non-degradable protein (NDP), potentially degradable protein proportion (PDP), and protein escaping rumen degradation (PEP) while decreasing water soluble protein (WSP) and the rates of crude protein (CP), except immature berseem clover and cell wall (CW) degradation. PEP was greater and rate of CP degradation was lower at 100 and 150o C compared to 125o C in immature berseem clover.

Comparison of inorganic nitrogen uptake dynamics following snowmelt and at peak biomass in subalpine grasslands

Subalpine grasslands are highly seasonal environments and likely subject to strong variability in nitrogen (N) dynamics. Plants and microbes typically compete for N acquisition during the growing season and particularly at plant peak biomass. During snowmelt, plants could potentially benefit from a decrease in competition by microbes, leading to greater plant N uptake associated with active growth and freeze-thaw cycles restricting microbial growth. In managed subalpine grasslands, we expect these interactions to be influenced by recent changes in agricultural land use, and associated modifications in plant and microbial communities. At several subalpine grasslands in the French Alps, we added pulses of 15N to the soil at the end of snowmelt, allowing us to compare the dynamics of inorganic N uptake in plants and microbes during this period with that previously reported at the peak biomass in July. In all grasslands, while specific shoot N translocation (per g of biomass) of dissolved inorganic nitrogen (DIN) was two to five times greater at snowmelt than at peak biomass, specific microbial DIN uptakes were similar between the two sampling dates. On an area basis, plant communities took more DIN than microbial communities at the end of snowmelt when aboveground plant biomasses were at least two times lower than at peak biomass. Consequently, inorganic N partitioning after snowmelt switches in favor of plant communities, allowing them to support their growing capacities at this period of the year. Seasonal differences in microbial and plant inorganic N-related dynamics were also affected by past (terraced vs. unterraced) rather than current (mown vs. unmown) land use. In terraced grasslands, microbial biomass N remained similar across seasons, whereas in unterraced grasslands, microbial biomass N was higher and microbial C : N lower at the end of snowmelt as compared to peak biomass. Further investigations on microbial community composition and their organic N uptake dynamics are required to better understand the decrease in microbial DIN uptake.

Herbivore identity mediates the strength of trophic cascades on individual plants

The strength of top-down indirect effects of carnivores on plants (trophic cascades) varies greatly and may depend on the identity of the intermediate (herbivore) species. If the effect strength is linked to functional traits of the herbivores then this would allow for more general predictions. Due to the generally sub-lethal effects of herbivory in terrestrial systems, trophic cascades manifest themselves in the first instance in the fitness of individual plants, affecting both their numerical and genetic contributions to the population. We directly compare the indirect predator effects on growth and reproductive output of individual Vicia faba plants mediated by the presence of two aphid species: Acyrtosiphon pisum is characterised by a boom and bust strategy whereby colonies grow fast and overexploit their host plant individual while Megoura viciae appear to follow a more prudent strategy that avoids over-exploitation and death of the host plant.Plants in the field were infested with A. pisum, M. viciae or both and half the plants were protected from predators. Exposure to predators had a strong impact on the biomass of individual plants and the strength of this effect differed significantly between the different herbivore treatments.A. pisum had a greater direct impact on plants and this was coupled with a significantly stronger indirect predator effect on plant biomass.Although the direct impact of predators was strongest on M. viciae, this was not transmitted to the plant level, indicating that the predator-prey interactions strength is not as important as the plant-herbivore link for the magnitude of the indirect predator impact. At the individual plant level, the indirect predator effect was purely due to consumptive effects on herbivore densities with no evidence for increased herbivore dispersal in response to presence of predators. The nature of plant-herbivore interactions is the key to terrestrial trophic cascade strength. The two herbivores that we compared were similar in feeding mode and body size but differed their way how they exploit host plants, which was the important trait explaining the strength of the trophic cascade.

Are ectomycorrhizal fungi alleviating or aggravating nitrogen limitation of tree growth in boreal forests?

•Symbioses between plant roots and mycorrhizal fungi are thought to enhance plant uptake of nutrients through a favourable exchange for photosynthates. Ectomycorrhizal fungi are considered to play this vital role for trees in nitrogen (N)-limited boreal forests. •We followed symbiotic carbon (C)–N exchange in a large-scale boreal pine forest experiment by tracing 13CO2 absorbed through tree photosynthesis and 15N injected into a soil layer in which ectomycorrhizal fungi dominate the microbial community. •We detected little 15N in tree canopies, but high levels in soil microbes and in mycorrhizal root tips, illustrating effective soil N immobilization, especially in late summer, when tree belowground C allocation was high. Additions of N fertilizer to the soil before labelling shifted the incorporation of 15N from soil microbes and root tips to tree foliage. •These results were tested in a model for C–N exchange between trees and mycorrhizal fungi, suggesting that ectomycorrhizal fungi transfer small fractions of absorbed N to trees under N-limited conditions, but larger fractions if more N is available. We suggest that greater allocation of C from trees to ectomycorrhizal fungi increases N retention in soil mycelium, driving boreal forests towards more severe N limitation at low N supply.

Effect of continuous gastric enteral feeding on tonometric measurement of gastric intramucosal carbon dioxide levels in healthy volunteers

Purpose. The aim of this research was to evaluate the effect of enteral feeding on tonometric measurement of gastric regional carbon dioxide levels (PrCO2) in normal healthy volunteers. Design and methods. The sample included 12 healthy volunteers recruited by the University Clinical Research Center (UCRC). An air tonometry system monitored PrCO2 levels using a tonometer placed in the lumen of the stomach via orogastric intubation. PrCO2 was automatically measured and recorded every 10 minutes throughout the five hour study period. An oral dose of famotidine 40 mg was self-administered the evening prior to and the morning of the study. Instillation of Isocal® High Nitrogen (HN) was used for enteral feeding in hourly escalating doses of 0, 40, 60, and 80 ml/hr with no feeding during the fifth hour. Results . PrCO2 measurements at time 0 and 10 minutes (41.4 ± 6.5 and 41.8 ± 5.7, respectively) demonstrated biologic precision (Levene's Test statistic = 0.085, p-value 0.774). Biologic precision was lost between T130 and T140 40 when compared to baseline TO (Levene's Test statistic = 1.70, p-value 0.205; and 3.205, p-value 0.042, respectively) and returned to non-significant levels between T270 and T280 (Levene's Test statistic = 3.083, p-value 0.043; and 2.307, p-value 0.143, respectively). Isocal® HN significantly affected the biologic accuracy of PrCO2 measurements (repeated measures ANOVA F 4.91, p-value <0.001). After 20 minutes of enteral feeding at 40 ml/hr, PrCO2 significantly increased (41.4 ± 6.5 to 46.6 ± 4.25, F = 5.4, p-value 0.029). Maximum variance from baseline (41.4 ± 6.5 to 61.3 ± 15.2, F = 17.22, p-value <0.001) was noted after 30 minutes of Isocal® HN at 80 ml/hr or 210 minutes from baseline. The significant elevations in PrCO2 continued throughout the study. Sixty minutes after discontinuation of enteral feeding, PrCO2 remained significantly elevated from baseline (41.4 ± 6.5 to 51.8 ± 9.2, F = 10.15, p-value 0.004). Conclusion. Enteral feeding with Isocal® HN significantly affects the precision and accuracy of PrCO2 measurements in healthy volunteers. ^

More efficient aboveground nitrogen use in more diverse Central European forest canopies

We hypothesized that biodiversity improves ecosystem functioning and services such as nutrient cycling because of increased complementarity. We examined N canopy budgets of 27 Central European forests of varying dominant tree species, stand density, and tree and shrub species diversity (Shannon index) in three study regions by quantifying bulk and fine particulate dry deposition and dissolved below canopy N fluxes. Average regional canopy N retention ranged from 16% to 51%, because of differences in the N status of the ecosystems. Canopy N budgets of coniferous forests differed from deciduous forest which we attribute to differences in biogeochemical N cycling, tree functional traits and canopy surface area. The canopy budgets of N were related to the Shannon index which explained 14% of the variance of the canopy budgets of N, suggesting complementary aboveground N use of trees and diverse understorey vegetation. The relationship between plant diversity and canopy N retention varied among regional site conditions and forest types. Our results suggest that the traditional view of belowground complementarity of nutrient uptake by roots in diverse plant communities can be transferred to foliar uptake in forest canopies.

Seasonal controls on grassland microbial biogeography: Are they governed by plants, abiotic properties or both?

Temporal dynamics create unique and often ephemeral conditions that can influence soil microbial biogeography at different spatial scales. This study investigated the relation between decimeter to meter spatial variability of soil microbial community structure, plant diversity, and soil properties at six dates from April through November. We also explored the robustness of these interactions over time. An historically unfertilized, unplowed grassland in southwest Germany was selected to characterize how seasonal variability in the composition of plant communities and substrate quality changed the biogeography of soil microorganisms at the plot scale (10 m x 10 m). Microbial community spatial structure was positively correlated with the local environment, i.e. physical and chemical soil properties, in spring and autumn, while the density and diversity of plants had an additional effect in the summer period. Spatial relationships among plant and microbial communities were detected only in the early summer and autumn periods when aboveground biomass increase was most rapid and its influence on soil microbial communities was greatest due to increased demand by plants for nutrients. Individual properties exhibited varying degrees of spatial structure over the season. Differential responses of Gram positive and Gram negative bacterial communities to seasonal shifts in soil nutrients were detected. We concluded that spatial distribution patterns of soil microorganisms change over a season and that chemical soil properties are more important controlling factors than plant density and diversity. Finer spatial resolution, such as the mm to cm scale, as well as taxonomic resolution of microbial groups, could help determine the importance of plant species density, composition, and growth stage in shaping microbial community composition and spatial patterns. (C) 2014 The Authors. Published by Elsevier Ltd. All rights reserved.