Micro-scaled high-throughput digestion of plant tissue samples for multi-elemental analysis

Background  

Quantitative multi-elemental analysis by inductively coupled plasma (ICP) spectrometry depends on a complete digestion of solid samples. However, fast and thorough sample digestion is a challenging analytical task which constitutes a bottleneck in modern multi-elemental analysis. Additional obstacles may be that sample quantities are limited and elemental concentrations low. In such cases, digestion in small volumes with minimum dilution and contamination is required in order to obtain high accuracy data.     

Herbivory of an invasive slug in a model grassland community can be affected by earthworms and mycorrhizal fungi

Invasion of non-native species is among the top threats for the biodiversity and functioning of native and agricultural ecosystems worldwide. We investigated whether the herbivory of the slug Arion vulgaris (formerly Arion lusitanicus; Gastropoda), that is listed among the 100 worst alien species in Europe, is affected by soil organisms commonly present in terrestrial ecosystems (i.e. earthworms—Annelida: Lumbricidae and arbuscular mycorrhizal fungi—AMF, Glomerales). We hypothesized that slug herbivory would be affected by soil organisms via altered plant nutrient availability and plant quality. In a greenhouse experiment, we created a simple plant community consisting of a grass, a forb, and a legume species and inoculated these systems with either two earthworm species and/or four AMF taxa. Slugs were introduced after plants were established. Earthworms significantly reduced total slug herbivory in AMF-inoculated plant communities (P?=?0.013). Across plant species, earthworms increased leaf total N and secondary metabolites, AMF decreased leaf thickness. Mycorrhizae induced a shift in slug feeding preference from non-legumes to legumes; the grass was generally avoided by slugs. AMF effects on legume herbivory can partly be explained by the AMF-induced increase in total N and decrease in C/N ratio; earthworm effects are less clear as no worm-induced alterations of legume plant chemistry were observed. The presence of earthworms increased average AMF colonization of plant roots by 140 % (P?<?0.001). Total shoot mass was significantly increased by AMF (P?<?0.001). These data suggest that the feeding behavior of this invasive slug is altered by a belowground control of plant chemical quality and community structure.     

Carbon isotopic ratios of wetland and terrace soil sequences in the Maya Lowlands of Belize and Guatemala

This article provides new data and synthesizes earlier findings on the carbon isotope ratios of the humin part of soil organic matter from a range of sites in the central Maya Lowlands. Changes down the soil profile in carbon isotope ratios can provide an important line of evidence for vegetation change and erosion over time, especially in well dated aggrading profiles. Research thus far has provided substantial evidence for significant inputs from C₄ vegetation in buried layers from the Ancient Maya periods in depositional soils but equivocal evidence from sloping soils. We present new findings from soil profiles through ancient Maya wetland fields, upland karst wetlands, ancient Maya aguadas (reservoirs), and ancient Maya terraces. Most of the profiles exhibited δ¹³C enrichment greater than the 2.5–3‰ typical from bacterial fractionation. Seven of nine ancient Maya wetland profiles showed δ¹³C enrichment ranging from 4.25 to 8.56‰ in ancient Maya-dated sediments that also contained phytolith and pollen evidence of grass (C₄ species) dominance. Upland karst sinks and ancient reservoirs produced more modest results for δ¹³C enrichment. These seasonal wetland profiles exhibited δ¹³C enrichment ranging from 1 to 7.3‰ from the surface to ancient Maya-period sediments. Agricultural terraces produced mixed results, with two terraces having substantial δ¹³C enrichment of 5.34 and 5.66‰ and two producing only equivocal results of 1.88 and 3.03‰ from modern topsoils to Maya Classic-period buried soils. Altogether, these findings indicate that C₄ plants made up c. 25% of the vegetation at our sites in the Maya Classic period and only a few percent today. These findings advance the small corpus of studies from ancient terraces, karst sinks, and ancient wetland fields by demonstrating substantial δ¹³C and thus C₄ plant enrichment in soil profile sections dated to ancient Maya times. These studies are also providing a new line of evidence about local and regional soil and ecological change in this region of widespread environmental change in the Late Holocene.     

Spatial modelling of the fraction of photosynthetically active radiation absorbed by a boreal mixedwood forest using a lidar–hyperspectral approach

The spatial variability of the fraction of photosynthetically active radiation absorbed by the canopy (fPAR) was characterized for a heterogeneous boreal mixedwood forest site located in northern Ontario, Canada, based on relationships found between fPAR and light detection and ranging (lidar) data over different canopy architectures. Estimates of fPAR were derived from radiation measurements made above the canopy at a flux tower and below-canopy radiation was measured across a range of species compositions and canopy architectures. Airborne lidar data were used to characterize spatial variability of canopy structure around the flux tower and a map of mean canopy chlorophyll concentration was derived from airborne hyperspectral imagery. Different volumes of lidar points for the locations directly above each photosynthetically active radiation (PAR) sensor were examined to determine if there is an optimal method of relating lidar returns to estimated fPAR values.The strongest correlations between mean lidar height and fPAR occurred when using points that fell within a theoretical cone which originated at the PAR sensor having a solid angle α = 55°. For diffuse conditions, the correlation (r) between mean lidar height versus fPAR × chlorophyll was stronger than between mean lidar height versus fPAR by 8% for mean daily fPAR and from 10 to 20% for diurnal fPAR, depending on solar zenith angle. For direct light conditions, the relationship was improved by 12% for mean daily fPAR and 12–41% for diurnal relationships.Linear regression models of mean daily fPAR × chlorophyll versus mean lidar height were used in conjunction with gridded lidar data and the canopy chlorophyll map to generate maps of mean daily fPAR for direct and diffuse sunlight conditions. Site average fPAR calculated from these maps was 0.79 for direct light conditions and 0.78 for diffuse conditions. When compared to point estimates of mean daily fPAR calculated on the tower, the average fPAR was significantly lower than the point estimate. Subtracting the direct sunlight fPAR map from the diffuse sunlight fPAR map revealed a distinct spatial pattern showing that areas with open canopies and relatively low chlorophyll (e.g., black spruce patches) have a higher fPAR under direct sunlight conditions, while closed canopies with higher chlorophyll (e.g., deciduous species) absorb more PAR under diffuse conditions. These findings have implications for scaling from point measurements at flux towers to larger resolution satellite imagery and addressing local scale heterogeneity in flux tower footprints.     

Soil properties and the macrofauna community in abandoned irrigated rice fields of northeastern Argentina

This study compared soil physical, chemical, and biological characteristics between natural grassland and recently abandoned rice fields in order to identify those variables that might explain the observed increase of Camponotus punctulatus anthills in abandoned rice paddy fields from Northern Argentina. Mainly due to a reduction of macropores and mesopores, overall porosity decreased by around 6% and bulk density was about 7% greater, in the 0- to 10- and 10- to 20-cm layers of the abandoned rice fields. Carbon and nitrogen content from organic matter increased (29% and 41% respectively for the 0- to 20-cm horizon) during cultivation but decreased (38% and 24%) 2 years after the last rice harvest. Forty percent of natural grassland-organic matter and 30% of abandoned rice-organic matter mineralized in less than 2 years. There was a different community structure between the abandoned rice fields and the undisturbed natural grassland and only a 20.6% (i.e. only 19 species from a total of 92) overlap in species composition. The abundance of macrofauna was greater in abandoned rice fields (2,208 individuals m^–2) in comparison to natural grasslands (288 ind m^–2) due to higher densities of small earthworms and Camponotus punctulatus ants; however, the Shannon index showed lower values in comparison to natural grasslands. Earthworms and C. punctulatus in the abandoned rice fields showed a change in their ¹³C signature indicating a switch in diet from natural grassland organic matter (C4) to organic matter from rice (C3). Our results indicate that the effects of rice cultivation practices did not seem to produce any physical or trophic limitations to recolonization by the macrofauna. It seems that changes in overall soil conditions have favored a change in the construction behavior of C. punctulatus which, in combination with population increases, could explain the explosion in number of anthills.     

Classification of Azolla spp., section Azolla

Summary Azolla accessions (section Azolla) from the germplasm collections of the International Rice Research Institute and Washington State University were fingerprinted and classified by enzyme electrophoresis and leaf trichome morphology. A. filiculoides was enzymatically distinctive and also reliably identified by its prominent one-celled trichomes. Neotropical accessions labelled as A. filiculoides proved to be members of other species. Two groups of isolates were designated A. rubra, but those from Japan were identified as A. filiculoides. The A. rubra of Australia-New Zealand was biochemically unique and possessed less protuberant trichomes than A. filiculoides. A. microphylla, A. mexicana, and A. caroliniana were phenetically similar, but a. microphylla was identifiable from the others in the banding patterns of certain enzymes. A. mexicana and A. caroliniana were closely related enzymatically. The two-celled leaf trichomes of these three species were similar in size and shape.     

Estimation of genetic variance components and heritabilities for cut-flower yield in gerbera using least squares and maximum likelihood methods

Summary Genetic variance components and heritability were estimated for cut-flower yield of gerbera in the Davis population using ordinary least squares and maximum likelihood methods. The overall estimate of narrow-sense heritability is 0.33 based on least squares (LS) and 0.31 based on maximum likelihood (ML). The results of the study indicate that (1) ML and LS provide very similar results if sample size is large enough, suggesting both are useful for plant breeding programs; (2) about one third of the variation in gerbera cut-flower yield is additive, implying selection in cut-flower will be successful; and (3) although additive variation gradually decreased, heritability remained near 0.27 suggesting there is still potential variation in the population for further selection.     

The use of lysimeter data for the test of two soil–water balance models: A case study

Two soil–water balance models were tested by a comparison of simulated with measured daily rates of actual evapotranspiration, soil water storage, groundwater recharge, and capillary rise. These rates were obtained from twelve weighable lysimeters with three different soils and two different lower boundary conditions for the time period from January 1, 1996 to December 31, 1998. In that period, grass vegetation was grown on all lysimeters. These lysimeters are located in Berlin‐Dahlem, Germany. One model calculated the soil water balance using the Richards equation. The other one used a capacitance approach. Both models used the same modified Penman formula for the estimation of potential evapotranspiration and the same simple empirical vegetation model for the calculation of transpiration, interception, and evaporation. The comparisons of simulated with measured model outputs were analyzed using the modeling‐efficiency index IA and the root mean squared error RMSE. At some lysimeters, the uncalibrated application of both models led to an underestimation of cumulative and annual rates of groundwater recharge and capillary rise, despite a good simulation quality in terms of IA and RMSE. A calibration of soil‐hydraulic and vegetation parameters such as maximum rooting depth resulted in a better fit between simulated and observed cumulative and annual rates of groundwater recharge and capillary rise, but in some cases also decreased the simulation quality of both models in terms of IA and RMSE. The results of this calibration indicated that, in addition to a precise determination of the soil water‐retention functions, vegetation parameters such as rooting depth should also be observed. Without such information, the rooting depth is a calibration parameter. However, in some cases, the uncalibrated application of both models also led to an acceptable fit between measured and simulated model outputs.     

Orosensory-directed identification of astringent mouthfeel and bitter-tasting compounds in red wine

Application of sequential solvent extraction, followed by HPLC combined with the taste dilution analysis, enabled the localization of the most intense velvety astringent, drying, and puckering astringent, as well as bitter-tasting, compounds in red wine, respectively. Isolation of the taste components involving gel adsorption chromatography, ultrafiltration, and synthesis revealed the identification of 26 sensory-active nonvolatiles, among which several hydroxybenzoic acids, hydroxycinnamic acids, flavon-3-ol glycosides, and dihydroflavon-3-ol rhamnosides as well as a structurally undefined polymeric fraction (>5 kDa) were identified as the key astringent components. In contradiction to literature suggestions, flavan-3-ols were found to be not of major importance for astringency and bitter taste, respectively. Surprisingly, a series of hydroxybenzoic acid ethyl esters and hydroxycinnamic acid ethyl esters were identified as bitter compounds in wine. Taste qualities and taste threshold concentrations of the individual wine components were determined by means of a three-alternative forced-choice test and the half-mouth test, respectively.     

Concept and feasibility study for the integrated evaluation of environmental monitoring data in forests

In the 1970s unexpected forest damages, called “new type of forest damage” or “forest decline”, were observed in Germany and other European countries. The Federal Republic of Germany and the German Federal States implemented a forest monitoring system in the early 1980s, in order to monitor and assess the forest condition. Due to the growing public awareness of possible adverse effects of air pollution on forests, in 1985 the ICP Forests was launched under the convention on long-range transboundary air pollution (CLRTAP) of the United Nations Economic Commission for Europe (UN-ECE). The German experience in forest monitoring was a base for the implementation of the European monitoring system. In 2001 the interdisciplinary case study “concept and feasibility study for the integrated evaluation of environmental monitoring data in forests”, funded by the German Federal Ministry of Education and Research, concentrated on in-depths evaluations of the German data of forest monitoring. The objectives of the study were: (a) a reliable assessment of the vitality and functioning of forest ecosystems, (b) the identification and quantification of factors influencing forest vitality, and (c) the clarification of cause-effect-relationships leading to leaf/needle loss. For these purposes additional data from external sources were acquired: climate and deposition, for selected level I plots tree growth data, as well as data on groundwater quality. The results show that in particular time series analysis (crown condition, tree growth, and tree ring analysis), in combination with climate and deposition are valuable and informative, as well as integrated evaluation of soil, tree nutrition and crown condition data. Methods to combine information from the extensive and the intensive monitoring, and to transfer process information to the large scale should be elaborated in future.