Molecular characterization of a stable antisense chalcone synthase phenotype in strawberry (Fragaria x ananassa)

An octaploid (Fragaria x ananassa cv. Calypso) genotype of strawberry was transformed with an antisense chalcone synthase (CHS) gene construct using a ripening related CHS cDNA from Fragaria x ananassa cv. Elsanta under the control of the constitutive CaMV 35S promoter via Agrobacterium tumefaciens. Out of 25 transgenic lines, nine lines showed a reduction in CHS mRNA accumulation of more than 50% as compared to the untransformed cv. Calypso control. The antisense CHS construct was found to be integrated into the genome, with a copy number ranging from one to four. The pigmentation of the fruit was only affected when less than 5% of the control CHS expression level was detected. A stable antisense phenotype over a period of 4 years was obtained in the primary transgenic lines at a rate of 1:20. As a consequence of the reduced activity of CHS, the levels of anthocyanins, flavonols, and proanthocyanidins were downregulated and precursors of the flavonoid pathway were shunted to the phenylpropanoid pathway leading to highly increased levels of cinnamoyl glucose (520% of control), caffeoyl glucose (816% of control), and feruloyl glucose (1092% of control) as well as p-coumaryl alcohol (363% of control) and p-coumaryl-1-acetate (1079% of control), which occur only as trace components in untransformed control fruits. These results demonstrate that the introduction of an antisense CHS construct in strawberry results in an unpredictable biochemical phenotype, thereby confirming that CHS function is an important regulatory point of substrate flow between the flavonoid and the phenylpropanoid pathways.     

Insight into the distribution of arabinoxylans, endoxylanases, and endoxylanase inhibitors in industrial wheat roller mill streams

To gain insight into the distribution of arabinoxylans (AX), endoxylanases, and endoxylanase inhibitors in industrial wheat roller milling, all streams, that is, 54 flour fractions, 4 bran fractions, and the germ, were analyzed for ash, starch, and protein contents, alpha-amylase activity levels, total (TOT-AX) and water-extractable arabinoxylan (WE-AX) contents, endoxylanase activity levels, and endoxylanase inhibitor (TAXI and XIP) contents. In general, bran fractions were significantly richer in TOT-AX and WE-AX contents, endoxylanase activity levels, and endoxylanase inhibitor contents than germ and, even more so, than flour fractions. In the 54 different flour fractions, minimal and maximal values for TOT-AX and WE-AX contents differed by ca. 2-fold, whereas they differed by ca. 15-fold for endoxylanase activity levels. The latter were positively correlated with ash and negatively correlated with starch content, suggesting that the endoxylanase activity in flour is strongly influenced by the level of bran contamination. TAXI contents in the flour fractions varied ca. 4-fold and were strongly correlated with bran-related parameters such as ash content and enzyme activity levels, whereas XIP contents varied ca. 3-fold and were not correlated with any of the parameters measured in this study. The results can be valuable in blending and optimizing wheat flour fractions to obtain flours with specific technological and nutritional benefits.     

Little effect of forest age on oribatid mites on the bark of trees

This study investigates the effect of forest age (20-, 50-, 160-year-old, and primeval forest) on oribatid mite communities on the bark of oak trees in the National Park Pusza Białowieża in eastern Poland. We hypothesized that oribatid mite diversity on bark peaks at forests of intermediate age and that the number of parthenogenetic species of oribatid mites is highest in young stands. In contrast to these hypotheses, the diversity, density, number of juveniles, community structure and the mode of reproduction of oribatid mite species were not significantly affected by forest age. None of the oribatid mite species occurred exclusively on trees of a specific age. The results suggest that oribatid mite communities on the bark of trees are minimally affected by tree harvesting regimes. In contrast to oribatid mites in soil, communities on bark appear to be less sensitive to disturbances.     

Die Oberflächenladung der Ton-Humusfraktion aus Schwarzerden als Funktion von pH und Ca-Konzentration

Surface charge of clay-humus fractions from Chernozems as a function of pH and Ca-concentration The charge characteristics of clay-humus fractions from A-horizons of several Chernozems as a function of Ca concentration and pH of the soil solution was investigated. The surface charge was measured by titration with charge compensating polyelectrolytes; the endpoint was at zero potential. A logarithmic dependence of the surface charge [mol_c/kg] on the Ca concentration of the soil solution in the range ? 2 mmol/L was observed. The influence of pH on the surface charge was almost linear over a wide range of pH-values, the change in pH-dependent charges being most pronounced for the horizons with low Ca saturation on the exchange sites. It was also found that the competition between protons and Ca ions for the exchange sites leads to a distinct decrease of the influence of Ca concentration on the surface charge in acid environment. The reason for slaking of the soil surface and of the displacement of clay-humus particles in Chernozems could be attributed to an increase in surface charge with decreasing Ca concentrations after leaching of lime. According to the present findings the drop in mobility of the clay-humus fractions during acidification could be explained by a charge compensating effect of protons and aluminium ions.     

Contribution of anecic earthworms to biopore formation during cultivation of perennial ley crops

Large sized biopores (diameter >2 mm) in the subsoil can be created by tap roots, which leave voids after their decay, or by the burrowing activity of anecic earthworms which may benefit from the temporary lack in tillage in perennial cropping systems. However, the interactions between root growth and earthworm activity in the process of biopore formation during perennial ley cropping are not well understood. The aim of this field study was to quantify the development of the abundance of the anecic earthworm Lumbricus terrestris and the biopore density during the cultivation of lucerne (Medicago sativa L.), chicory (Cichorium intybus L.) and tall fescue (Festuca arundinacea Schreb.) grown for either one, two or three years. An increased abundance of L. terrestris was already recorded after two years of continuous ley when compared with one year cultivation. The ley crop species had only minor influence on the abundance of L. terrestris. Biopore densities of both diameter classes under study (2–5 mm and >5 mm) were not significantly affected by the duration of ley cropping. In contrast, biopore densities were influenced by ley crop species. More biopores of diameter class 2–5 mm were recorded after chicory than after fescue. Lucerne cropping resulted in intermediate biopore density. Additionally, in an incubation experiment under field conditions, we quantified whether L. terrestris preferentially created new biopores or colonized abandoned, previously existing ones. After three weeks of incubation, one third of the adult individuals incubated in the experimental area created new biopores at 0.4 m soil depth. A similar percentage of individuals colonized previously existing biopores, partially widening the lumen of smaller sized biopores. The remaining individuals remained in the topsoil. Sub-adult individuals rarely formed new pores. Half of the introduced sub-adults remained in the topsoil. We conclude that in crop rotations new biopores can be generated during perennial ley cropping by taproot systems of ley crops, but that a two to three- year period without tillage is not sufficient for populations of anecic earthworms to make a marked contribution to biopore density in the subsoil. The relevance of anecic earthworms for altering physical and chemical properties of biopores during ley cropping still needs further investigation.     

Aspects of phytoremediation of organic pollutants

Phytoremediation is a quite novel technique to clean polluted soils using plants. In theory, phytoremediation methods are cheap, are accepted by the public and, compared to physical or chemical approaches, are ecologically advantageous. Until today, however, there are only a few examples of successful applications. One reason is that the processes involved are complex, and a full clean up may require many years. Plants affect the water balance of a site, they change redox potential and pH, and stimulate microbial activity of the soil. These indirect influences may accelerate degradation in the root zone or reduce leaching of compounds to groundwater. Compounds taken up into plants may be metabolised, accumulated, or volatilised into air. Based on these processes, several phytoremediation methods have been developed: Phytoextraction, rhizofiltra-tion, phytostabilisation, rhizo and phytodegradation, pump and tree, land farming, phytovolatilisation, hydraulic control and more. Already in use are plants (and here willow, poplar and grass) for the degradation of petroleum products, aromatic hydrocarbons (BTEX), chlorinated solvents, explosives and cyanides. However, phytotoxicity and pollutant mass balances were rarely documented. Often, the success of the projects was not controlled, and only estimates can be made about the applicability and the potential of phytoremediation. This lack of experience about possibilities and limitations seems to be a hindrance for a broader use of these techniques.     

Quantitative Mineralbestimmung in der Schlufffraktion von Böden auf der Grundlage der chemischen Analyse und unter Anwendung der Karl-Fischer-Titration. I. Verfahren

Quantitative estimation of the mineralogical composition of silt fractions of soils based on both, chemical analysis and application of Karl-Fischer-titration. I. Method This paper describes a method for calculating the mineralogical composition of the silt fractions of soils. The chemical analysis and the temperature dependence of water release are used for determining micas and feldspars, each in three components in a relatively short procedure. The release of water was measured by Karl-Fischer-titration at controlled temperatures. At temperatures above 550°C the water release of the particle size fractions correlates with their content of micas. The quantity of micas can be obtained from the amount of released water and the results of the chemical analysis. The influence of kaolinite, amphibole and the fluoride content on the calculation is described. By considering the potassium content of the mica fraction results of the chemical analysis are used to quantify feldspars.     

Statistical Modeling of the Partitioning of Nonylphenol in Soil

Partition coefficients K _P of nonylphenol (NP) in soil were determined for 193 soil samples which differed widely in content of soil organic carbon (SOC), hydrogen activity, clay content, and in the content of dissolved organic carbon (DOC). By means of multiple linear regression analysis (MLR), pedotransfer functions were derived to predict partition coefficients from soil data. SOC and pH affected the sorption, though the latter was in a range significantly below the pK_a of NP. Quality of soil organic matter presumably plays an important but yet not quantified role in sorption of NP. For soil samples with SOC values less than 3 g kg^?1, model prediction became uncertain with this linear approach. We suggest that using only SOC and pH data results in good prediction of NP sorption in soils with SOC higher than 3 g kg^?1. Considering the varying validity of the linear model for different ranges of the most sensitive parameter SOC, a more flexible, nonlinear approach was tested. The application of an artificial neuronal network (ANN) to predict sorption of NP in soils showed a sigmoidal relation between K _P and SOC. The nonlinear ANN approach provided good results compared to the MLR approach and represents an alternative tool for prediction of NP partitioning coefficients.     

Use of isotope-labeled aflatoxins for LC-MS/MS stable isotope dilution analysis of foods

Aflatoxins are a group of very carcinogenic mycotoxins that can be found on a wide range of food commodities including nuts, cereals, and spices. In this study, the first LC-MS/MS stable isotope dilution assay (SIDA) for the determination of aflatoxins in foods was developed. The development of this method was enabled by easily accessible isotope-labeled (deuterated) aflatoxins B2 and G2, which were synthesized by catalytic deuteration of aflatoxin B1 and G1, purified, and well-characterized by NMR and MS. All four aflatoxins of interest (B1, B2, G1, and G2) were quantified in food samples by using these two labeled internal standards. The response factors (RF) of the linear calibrations were revealed to be matrix independent for labeled aflatoxin B2/aflatoxin B2 and labeled aflatoxin G2/aflatoxin G2. For labeled aflatoxin B 2/aflatoxin B 1 and labeled aflatoxin B2/aflatoxin G1 matrix-matched calibration was performed for the model matrices almonds and wheat flour, showing significant differences of the RFs. Limits of detection (LOD) were determined by applying a statistical approach in the presence of the two model matrices, yielding 0.31 microg/kg (aflatoxin B1), 0.09 microg/kg (aflatoxin B2), 0.38 microg/kg (aflatoxin G1), and 0.32 microg/kg (aflatoxin G2) for almonds (similar LODs were obtained for wheat flour). Recovery rates were between 90 and 105% for all analytes. Coefficients of variation (CV) of 12% (aflatoxin B1), 3.6% (aflatoxin B2), 14% (aflatoxin G1), and 4.8% (aflatoxin G2) were obtained from interassay studies. For further validation, a NIST standard reference food sample was analyzed for aflatoxins B1 and B2. The method was successfully applied to determine trace levels of aflatoxins in diverse food matrices such as peanuts, nuts, grains, and spices. Aflatoxin contents in these samples ranged from about 0.5 to 6 microg/kg.