Field evaluation of antisense RNA mediated inhibition of GBSS gene expression in potato

Summary Granule-bound starch synthase (GBSS) catalyses the synthesis of amylose in starch granules. Analysis of antisense RNA mediated inhibition of GBSS gene expression in large numbers of tubers from in vitro grown, greenhouse grown and field grown transgenic potato plants revealed stable and total inhibition of GBSS gene expression in one clone. In three other transgenic genotypes partial and unstable inhibition was found. In these genotypes both GBSS activity and amylose content were remarkably reduced compared with the non-transformed control genotype. No relationship was found between the level of inhibition of GBSS gene expression and yield and dry matter content.     

Impact of lakeshore development on green frog abundance

Many amphibian species exhibit metapopulation spatial dynamics and temporally are faced with local population extinction and re-colonization. These natural population fluctuations can exhibit stochastic effects when human-caused alteration and fragmentation of habitats occur during sensitive life-cycle events. In this study, we explored the effects of shoreline development on adult green frogs Rana clamitans melanota on lakes (n=24) of northern Wisconsin. We estimated green frog abundance using both auditory and direct observation surveys. The immediate shoreline habitat was mapped and placed into a Geographical Information System (GIS) for analysis. Adult green frog populations were significantly lower on lakes with varying degrees of shoreline house and cottage development than lakes with little or no development. A negative linear relationship existed between shoreline development densities and the number of adult green frogs. However, house and cottage densities alone did not directly explain this reduction. Analysis of variance (ANOVA) identified that the amount of suitable habitat, not development density, significantly affected green frog abundance. Therefore, greater development densities significantly decrease breeding habitat quality, resulting in lower adult frog abundance. These and other findings suggest that lakeshore development regulations are not protecting sensitive amphibian species.     

Der Bulk-Stomata-Widerstand von Ackerbohnen und seine Abhängigkeit vom Bodenwassergehalt

Bulk stomatal resistance of Vicia faba and its dependence on soil water content Bulk stomatal resistance of beans (Vicia faba L.,) as a function of soil water content was estimated for 41 days by comparison of potential transpiration, determined from meteorological data, and actual transpiration, determined from soil water losses. The resistance of beans is about 46 s/m if soil water content is above 70 % of available field capacity and increases strongly in dryer situations. This behaviour can be described by an exponential function.     

Numerical taxonomic studies in the generaTriticum L. andPisum L.

Summary Up till now numerical taxonomy has been applied to cultivated plants only to a limited extent though this method seems to be especially suitable for handling the great diversity of these plants. Moreover the theory of numerical taxonomy has not been studied sufficiently.Therefore studies with cultivated plants were begun with the aim to investigate the fitness of different numerical taxonomic techniques for their classification. Materials used: 1) 24 species ofTriticum and additionally 24 species ofAegilops 2) 1225Triticum taxa from Iran (mostlyT. aestivum) 3) 312 taxa ofPisum (mostlyP. sativum).Using 10 coefficients and 7 clustering methods inTriticum andAegilops in most cases a clear separation of both genera resulted as well as a species arrangement withinTriticum according to the genomes.The main aim when dealing with the wheat from Iran was the investigation of the influence of different character sets. At this the coefficient ofGower and the unweighted pair-group method using arithmetic averages (UPGMA) were applied. The separation of the different species is satisfying clear while the representatives of the different varieties ofT. aestivum could not always be found in one cluster. This effect increased when adding further characters sets.InPisum the influence of 11 similarity coefficients to the dendrogram formation has been investigated. With two-state characters the simple matching coefficient and with mainly multistate characters the Canberra metric gave best results (using UPGMA and information measures SUMRAT and SAMRAT).The hitherto existing results show the complicatedness of the evaluation of numerical taxonomic methods in their application to cultivated plants. Therefore generalizations are not yet possible.

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Numerisch-taxonomische Studien in den GattungenTriticum L. undPisum L.

     

Diurnal Macronutrients Uptake Patterns by Lettuce Roots under Various Light and Temperature Levels

In order to reduce nutrient wastes to the environment the supply should be in accordance to the demand for these. Two experiments were conducted to study and quantify the effect of temperature, irradiance, and plant age on the uptake of nitrate (NO₃^?), ammonium (NH₄⁺), dihydrogen phosphate ion (H₂PO₄^?), potassium (K⁺), calcium (Ca²⁺), magnesium (Mg²⁺), and sulfate (SO₄²). In the first experiment, various levels of temperature and irradiance were applied to plants in a growth chamber, while in the second experiment the uptake was studied along the crop season under greenhouse conditions. The uptake rates were calculated at 2-hour intervals through sampling the nutrient solution and analyzing it by inductively coupled plasma atomic emission spectrometry (ICP-AES). Increasing light and temperature enhance the uptake rates, while the rates decrease with plant age. Nitrogen absorption was similar during the day as during the night. No differences were found in the absorption of H₂PO₄^?, K⁺, Ca²⁺, Mg²⁺, and SO₄^2? between day and night. Nitrate absorption was found to have a positive correlation with the absorption of all the ions except for NH₄⁺.     

Pre-extraction preparation (fresh, frozen, freeze-dried, or acetone powdered) and long-term storage of fruit and vegetable tissues: effects on antioxidant enzyme activity

Activities of the antioxidant enzymes ascorbate peroxidase, catalase, dehydroascorbate reductase, glutathione reductase, guaiacol peroxidase, monodehydroascorbate reductase, and superoxide dismutase were assayed in honeydew (Cucumis melo L.) fruit and spinach (Spinacia oleracea L.) leaves either as fresh, frozen to -80 degrees C, frozen in liquid nitrogen, freeze-dried, or acetone powder, representing the various ways tissues are treated prior to enzyme extraction. Treated tissues were analyzed following treatment or stored for up to 8 weeks at -80 degrees C. Enzyme activities in fruit frozen with or without liquid nitrogen and leaves frozen with or without liquid nitrogen or freeze-dried were equal to those of fresh tissue. Enzyme activities in freeze-dried or acetone-powdered fruit and leaves and in acetone-powdered tissues were significantly higher or lower than those in fresh tissue. Enzyme activities in both tissues frozen with or without liquid nitrogen and stored for 8 weeks at -80 degrees C changed little; those in freeze-dried and acetone-powdered tissues, however, significantly increased/decreased over the same period. Fresh tissue should be used in antioxidant enzyme assays, but if storage is necessary, tissues should be placed directly into a -80 degrees C freezer.     

Bioassay-directed Fractionation of Organic Extracts of Marine Surface Sediments from the North and Baltic Sea - Part II: Results of the biotest battery and development of a biotest index (8 pp)

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Part I: Determination and identification of organic pollutants Part II: Results of the biotest battery and development of a biotest index

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Preamble. This series of two papers presents the results of an interdisciplinary research project (ISIS) dealing with bioassay-directed fractionation of marine sediment extracts. Part I presents the extraction and fractionation procedure as well as the results of chemical analysis, including non-target analysis of sediments. Part II describes the results of the biotest battery in relation to chemicals possibly causing parts of the observed effects. A biotest index is used to compare the toxicities of the samples.

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AUTHORS / AFFILIATIONS Ninja Reineke (3), Werner Wosniok (4), Dirk Danischewski (1), Heinrich Hühnerfuss (3), Angelika Kinder (5), Arne Sierts-Herrmann (5), Norbert Theobald (2), Hans-Heinrich Vahl (6), Michael Vobach (1), Johannes Westendorf (6) and Hans Steinhart (5).

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(1) Federal Research Centre for Fisheries, Institute for Fishery Ecology, Palmaille 9, 22767 Hamburg, Germany (2) Federal Maritime and Hydrographic Agency, Bernhard-Nochtstr. 78, 20359 Hamburg, Germany (3) University of Hamburg, Institute for Organic Chemistry, Martin-Luther-King-Platz 6, 20146 Hamburg, Germany (4) University of Bremen, Institute of Statistics, Bibliothekstr. 1, 28334 Bremen, Germany (5) University of Hamburg, Institute for Food Chemistry, Grindelallee 117, 20146 Hamburg, Germany (6) University of Hamburg, University Hospital Hamburg-Eppendorf, Department for Toxicology, Vogt-Kölln-Str. 30, 22527 Hamburg, Germany (7) Eurofins Wiertz-Eggert-Jörissen, Stenzelring 14b, 21107 Hamburg, Germany

Goal, Scope and Background

The ecological relevance of contaminants in mixtures is difficult to assess, because of possible interactions and due to lacking toxicity data for many substances present in environmental samples. Marine sediment extracts, which contain a mixture of environmental contaminants in low concentrations, were the object of this study. The extracts were investigated with a set of different biotests in order to identify the compound or the substance class responsible for the toxicity. For this goal, a combination of biotests, biotest-directed fractionation and chemical analysis has been applied. Further on, a strategy for the development of a biotest index to describe the toxicity of the fractions without a prior ranking of the test results is proposed. This article (Part II) focuses on the biological results of the approach.

Methods

The toxicological potential of organic extracts of sediments from the North Sea and the Baltic Sea was analyzed in a bioassay-directed fractionation procedure with a set of biotests: luciferase reporter gene assays on hormone receptor and Ah receptor, arabinose resistance test, fish embryo test (Danio rerio), comet assay, acetylcholinesterase inhibition test, heat-shock protein 70 induction, oxidative stress and luminescence inhibition test (Vibrio fischeri). The test results provided the basis for the calculation of a biotest index by factor analysis to compare the toxicity of the samples and fractions.

Results and Discussion

Results of 11 biotests on different fractionation levels of the samples were described and discussed with regard to the occurrence of contaminants and their toxic potentials. Polychlorinated biphenyls, polycyclic aromatic hydrocarbons, quinones, brominated indoles and brominated phenols were in the focus of interest. A biotest index was constructed to compare the toxic responses in the samples and to group the biotest results.

Conclusion

The procedure presented in this study is well suited for bioassay-directed fractionation of marine sediment extracts. However, in relatively low contaminated samples, high enrichment factors and sufficient fractionation is necessary to allow identification of low concentrations of contaminants which is required to link effects and possible causes. In the present case, the relation between substances and effects was difficult to uncover due to relatively low concentrations of pollutants compared to the biogenic matrix and to the remaining complexity of the fractions. The results, with respect to the brominated phenols and indoles in the samples, highlight the successful use of bioassay directed fractionation in the case of high concentrations and high toxicity.

Recommendation and Outlook

In general, it has been shown that a marine risk assessment requires focusing on the input of diffuse sources and taking into account the fact of mixture toxicity. Effects resulting from biogenic substances will make the assessment of the influence of anthropogenic substances even more difficult.     

Release of nonexchangeable NH4-N after planting of ryegrass in relation to soil content and as affected by nitrate supply

The uptake of N by ryegrass grown in pot culture on a range of soils differing widely in content of nonexchangeable NH₄-N (topsoils: 117 to 354 mg kg^?1 soil; subsoils: 117 to 270 mg kg^?1 soil) was measured to indicate whether the amounts of NH₄-N released from clay minerals were correlated with soil NH₄-N. After two cuts soil analysis revealed that the amounts of mobilized nonexchangeable NH₄-N were between 3.5 and 25.2 mg kg^?1 from topsoils and between 0 and 8.2 mg kg^?1 from subsoils. There was no correlation between soil nonexchangeable NH₄-N content and release. The NH₄-N extracted with 1 N HCl and the actual N uptake of the plants correlated highly significant. Assuming that the whole of the NH₄-N released was taken up by ryegrass, NH₄-N accounted for 11.2 to 75.0% of total N uptake from topsoils and 0 to 37.3% from subsoils. The release of nonexchangeable NH₄-N was increased by the application of nitrate.