Extending the working range of immunoanalysis by exploitation of two monoclonal antibodies

A newly established rat monoclonal antibody (mAb) for isoproturon, namely, IOC 10G7, is described. This mAb shows a standard curve for isoproturon in phosphate-buffered saline with a test midpoint of 5.5 +/- 1.8 microg/L (n = 15). In combination with the formerly developed mAb IOC 7E1, IOC 10G7 can be exploited to extend the working range for the analysis of isoproturon. Both antibodies were formatted into a competitive enzyme-linked immunosorbent assay (ELISA), using the same enzyme-tracer. MAb IOC 7E1 and mAb IOC 10G7 have different affinities for the target compound, but the signal-response curves of the single antibodies overlap. Cross-reactivity (CR) patterns of both antibodies are comparable, showing the highest CR for the metabolite 1-(4-isopropylphenyl)-3-methylurea (IOC 10G7, 9%; IOC 7E1, 19%). The system described here includes the combined, but individual, usage of both assays on one microtiter plate, as well as the strategy for mixing the two antibodies for the utilization in one assay. When standards are performed in Milli-Q water, the working range for isoproturon with the individual ELISAs using mAb IOC 7E1 is from 0.01 to 1 microg/L (test midpoint = 0.11 +/- 0.03 microg/L; n = 17) and with IOC 10G7, it is 1-100 microg/L (test midpoint = 10.3 +/- 1.6 microg/L; n = 32). The working range with mixed antibodies is usually on the order of 0.03-30 microg/L (test midpoint = 0.5 +/- 0.2 microg/L; n = 17). These strategies (mAbs individually and mixed) cover a range of 4 and 3 orders of magnitude, respectively. As a demonstration, water samples of different origins and an extract of mixed sediment were analyzed. The advantages of these strategies are discussed.     

Crystal structure of a photolyase bound to a CPD-like DNA lesion after in situ repair

DNA photolyases use light energy to repair DNA that comprises ultraviolet-induced lesions such as the cis-syn cyclobutane pyrimidine dimers (CPDs). Here we report the crystal structure of a DNA photolyase bound to duplex DNA that is bent by 50 degrees and comprises a synthetic CPD lesion. This CPD lesion is flipped into the active site and split there into two thymines by synchrotron radiation at 100 K. Although photolyases catalyze blue light-driven CPD cleavage only above 200 K, this structure apparently mimics a structural substate during light-driven DNA repair in which back-flipping of the thymines into duplex DNA has not yet taken place.     

Simple linear relationship between dry matter,specific gravity,and tissue specific gravity in a diploid potato breeding population

Summary Two hundred clones were randomly selected in 1988 and in 1989 from a hybrid population ofSolanum phureja andSolanum stenotomum to determine the relationship between specific gravity (SG) and percent dry matter (%DM), and between tissue specific gravity (TSG) and %DM. The objectives were: (1) to determine the dry matter prediction equation for a diploid potato population, (2) to determine whether estimates of %DM and efficiency of selection for %DM could be improved by measuring TSG, and (3) to determine the influence of year on the estimates. In 1988 the prediction equation for the relationship between %DM and SG was %DM=198.71 (SG-191.30,r=0.85. Ranges and means for the variables were: (1) %DM: 16.99–32.67, mean 25.16; (2) SG: 1.054–1.122, mean 1.089. In 1989 the relationship of %DM and SG was predicted as %DM=194.46(SG)-187.53,r=0.87; whereas the relationship of %DM and TSG was predicted as %DM=196.77(TSG)-192.91,r=0.87. Ranges and means for the variables were: (1) %DM: 10.68–34.57, mean 21.32; (2) SG: 1.030–1.122, mean 1.074; (3) TSG: 1.046–1.141, mean 1.089. Slopes were not significantly different between any of the prediction equations, but intercepts were significantly different. Estimates of %DM were not improved by measuring TSG. Year did influence the relationship between %DM and SG.     

Effects of continuous optimal fertilization on belowground ectomycorrhizal community structure in a Norway spruce forest

Studies of effects of fertilizer treatment on ectomycorrhizal fungal community structure have predominantly been based on large, single additions of nitrogen. Studies involving chronic additions of nutrients in combination with irrigation are much less common. We used morphotyping to study effects of balanced additions of a nutrient solution on ectomycorrhizal fungal community structure in a 36-year-old stand of Picea abies (L.) Karst. Despite high variability among individual samples, principal components analysis revealed a clear shift in community structure in response to fertilization. Irrigated plots receiving only water did not differ significantly from untreated control plots. Mycorrhizal root tips colonized by Cenococcum geophilum Fr. were significantly more common in fertilized plots than in control plots. Possible responses by other ectomycorrhizal species were masked by high variability. Over sixty morphotypes were distinguished, but there was no measurable effect of either fertilizer or irrigation treatment on morphotype richness or total number of root tips.     

Effect of varied soil nitrogen supply on growth and nutrient uptake of young Norway spruce plants grown in a shaded environment

To understand the effect of increased soil N supply on tree growth and nutrient uptake, three-year-old Norway spruce seedlings were grown in pots on low-nutrient mineral forest soil supplemented with N in mineral or organic form. Outdoor shaded growth conditions were used, to test the hypothesis that shaded plants are particularly susceptible to high soil N supply. Plants were harvested eleven months after planting. Shoot growth was not affected by the N supply, but N concentrations in needles and roots were increased in plants supplied with mineral N (150 or 300 mg N [kg soil]^—1). Root growth was drastically reduced and root/shoot ratios were decreased in plants with higher N uptake. A high supply of mineral N to soil also decreased the concentrations of other essential elements (P, K) in the needles and thus had effects on plant growth which may impair the stress resistance of trees. Organic N in the form of keratin (150 mg N [kg soil]^—1) did not influence plant growth significantly. The adverse effects of high mineral N supply were particularly pronounced under shaded conditions in comparison to results from other experiments using higher light intensity and temperature conditions.     

Detection and quantification of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis> in irrigation waters

A quantitative RT real-time PCR method was developed for the detection and quantification of Tomato mosaic virus (ToMV) in irrigation waters. These have rarely been monitored for the presence of plant pathogenic viruses, mostly due to the lack of efficient and sensitive detection methods. The newly developed method presented here offers a novel approach in monitoring the health status of environmental waters. ToMV was reliably detected at as low as 12 viral particles per real-time PCR reaction, which corresponds to the initial concentration of approximately 4.2 × 10^?10 mg (6,300 viral particles) of ToMV per ml of sample. The sensitivity of the method was further improved by including the Convective Interaction Media^® (CIM) monolithic chromatographic columns for quick and efficient concentration of original water samples. Seven out of nine water sources from different locations in Slovenia tested positive for ToMV, after concentrating the sample. Four samples tested ToMV-positive without the concentrating procedure. The presence and integrity of infective ToMV particles in the original sample, as well as in the chromatographic fraction, was confirmed using different methods from test plants, DAS ELISA to electron microscopy and real-time PCR. In this study, we propose a unique and simple diagnostic scheme for rapid, efficient, and sensitive monitoring of irrigation waters that could also be adopted for other plant, human or animal viruses.     

Untersuchungen zur Übertragung von Clavibacter michiganensis ssp. sepedonicus, dem Erreger der Bakteriellen Ringfäule, in das Kraut von Kartoffelpflanzen

From 1999 to 2001, field trials were carried out to find out weather there is a risk of the causal agent of Bacterial Ring Rot of potato (Clavibacter michiganensis ssp. sepedonicus) to be transmitted to the leaves of healthy potato plants via contaminated machinery and equipment during the vegetation period and if so weather the bacteria are transported into the newly formed tubers. Two methods were chosen. Firstly, a sharp-edged metal bar contaminated with a bacterial suspension was drawn through the leaves of potato plants up to 6 times during the vegetation period. Secondly, a suspension of the pathogen was directly injected into the stems of potato plants. The harvested tubers were tested in all trials, leaves were additionally tested in the trial with the contaminated metal bar. The pathogen was not found in any trial. Therefore the risk for the transmission of the Ringrot agent to the leaves of potato plants with contaminated machinery might be very low.     

Die neue Datenbank ALPS-BBA — der komfortable, schnelle Weg zu Informationen über Alternativen zum chemischen Pflanzenschutz für Erzeuger und Berater

With the database ALPS-BBA (http://ALPS.BBA.de), the Federal Biological Research Centre for Agriculture and Forestry (BBA) contributes to promoting integrated pest management and to the necessary extent of applying plant protection products in crop farms. ALPS-BBA offers a comfortable way to search for combinations of crops and pests, possible specific measures, and for key words. Running free text search is also possible. The free text search, in particular, has a great potential for random queries. Web front end and layout allow intuitive navigation, and one can say that search is barrier-free. The database provides information for users who are able to determine crops and pests. Therefore diagnostic aids are not included. Specialised scientists will permanently improve and update the database.     

The effects of metribuzin on early life stages of common carp (Cyprinus carpio)

Metribuzin belongs to the triazine group of herbicides, which are frequently used in agriculture. The aim of this study was to assess the impact of metribuzin on growth and development of early life stages of fish. Subchronic toxic effects of metribuzin at concentrations of 0.9, 4, 14, and 32 mg/L on embryos and larvae of common carp (Cyprinus carpio) were investigated during a 30 day toxicity test under experimental conditions. Exposure to metribuzin at 32 mg/L was associated with increased mortality. Negative effects on total body length, weight, and inhibition of specific growth rate were induced by all experimental concentrations. Length and weight parameters were the most sensitive. The negative impact of metribuzin was observed in the highest tested concentration beginning on day 6 of exposure. Retardation of early ontogeny was associated with concentrations ?4 mg/L. Histological examination revealed changes in liver and caudal kidney after 30 days exposure to 32 mg/L. Based on growth parameters, development, and histological examination, the Lowest Observed Effect Concentration (LOEC) value was 0.9 mg/L.