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Ecotoxicology of Organotin compounds

Organotin and especially butyltin compounds are used for a variety of applications, e.g. as biocides, stabilizers, catalysts and intermediates in chemical syntheses. Tributyltin (TBT) compounds exhibit the greatest toxicity of all organotins and have even been characterized as one of the most toxic groups of xenobiotics ever produced and deliberately introduced into the environment. TBT is not only used as an active biocidal compound in antifouling paints, which are designed to prevent marine and freshwater biota from settlement on ship hulls, harbour and offshore installations, but also as a biocide in wood preservatives, textiles, dispersion paints and agricultural pesticides. Additionally, it occurs as a by-product of mono- (MBT) and dibutyltin (DBT) compounds, which are used as UV stabilizer in many plastics and for other applications. Triphenyltin (TPT) compounds are also used as the active biocide in antifouling paints outside Europe and furthermore as an agricultural fungicide since the early 1960s to combat a range of fungal diseases in various crops, particularly potato blight, leaf spot and powdery mildew on sugar beet, peanuts and celery, other fungi on hop, brown rust on beans, grey moulds on onions, rice blast and coffee leaf rust. Although the use of TBT and TPT was regulated in many countries world-wide from restrictions for certain applications to a total ban, these compounds are still present in the environment. In the early 1970s the impact of TBT on nontarget organisms became apparent. Among the broad variety of malformations caused by TBT in aquatic animals, molluscs have been found to be an extremely sensitive group of invertebrates and no other pathological condition produced by TBT at relative low concentrations rivals that of the imposex phenomenon in prosobranch gastropods speaking in terms of sensitivity. TBT induces imposex in marine prosobranchs at concentrations as low as 0,5 ng TBT-Sn/L. Since 1993, for the littorinid snail Littorina littorea a second virilisation phenomenon, termed intersex, is known. In female specimens affected by intersex the pallial oviduct is transformed of towards a male morphology with a final supplanting of female organs by the corresponding male formations. Imposex and intersex are morphological alterations caused by a chronic exposure to ultra-trace concentrations of TBT. A biological effect monitoring offers the possibility to determine the degree of contamination with organotin compounds in the aquatic environment and especially in coastal waters without using any expensive analytical methods. Furthermore, the biological effect monitoring allows an assessment of the existing TBT pollution on the basis of biological effects. Such results are normally more relevant for the ecosystem than pure analytical data. usw.

Barley dwarfs acting big in agronomy. Identification of genes and characterization of proteins involved in dwarfism, lodging resistance and crop yield

Barley (Hordeum vulgare) is an important cereal grain which serves as major animal fodder crop as well as basis for malt beverages or staple food. Currently barley is ranked fourth in terms of quantity of cereal crops produced worldwide. In times of a constantly growing world population in conjunction with an unforeseeable climate change and groundwater depletion, the accumulation of knowledge concerning cereal growth and rate of yield gain is important. The Nordic Genetic Resource Center holds a major collection of barley mutants produced by irradiation or chemical treatment. One phenotypic group of barley varieties are dwarf mutants (erectoides, brachytic, semidwarf, uzu). They are characterized by a compact spike and high rate of yield while the straw is short and stiff, enhancing the lodging resistance of the plant. Obviously they are of applied interest, but they are also of scientific interest as virtually nothing is known about the genes behind the development of plant dwarfism. The aim of this project is to identify and isolate the genes carrying the mutations by using state of the art techniques for gene cloning at the Carlsberg Laboratory. The identified genes will be connected with the mutant phenotype to reveal the gene function in general. One or two genes will be overexpressed and the resulting recombinant proteins will be biochemically and structurally characterized. The insights how the mutation effects the protein will display the protein function in particular. Identified genes and their mutant alleles will be tested in the barley breeding program of the Carlsberg brewery.

Drivers and mechanisms of 13C discrimination in Cleistogenes squarrosa (C4) - reducing uncertainties on bundle sheath leakiness

The energetic efficiency of C4 photosynthesis is strongly affected by bundle sheath leakiness, which is commonly assessed with the 'linear version' of the Farquhar model of 13C discrimination, and leaf gas exchange and 13C composition data. But, the linear Farquhar model is a simplification of the full mechanistic theory of ? in C4 plants, potentially generating errors in the estimation of leakiness. In particular, post-photosynthetic C isotope fractionation could cause large errors, but has not been studied in any detail. The present project aims to improve the understanding of the ecological and developmental/physiological factors controlling discrimination and leakiness of the perennial grass Cleistogenes squarrosa. C. squarrosa is the most important member of the C4 community which has spread significantly in the Mongolia grasslands in the last decades. It has an unusually high and variable discrimination, which suggests very high (and potentially highly variable) leakiness. Specifically, we will conduct the first systematic study of respiratory 13C fractionation in light and dark at leaf- and stand-scale in this C4 species, and assess its effect on discrimination and estimates of leakiness. These experiments are conducted in specialized 13CO2/12CO2 gas exchange mesocosms using ecologically relevant scenarios, testing specific hypotheses on effects of environmental drivers and plant and leaf developmental stage on discrimination and leakiness.

Quantification of the influence of current use fungicides and climate change on allochthonous Organic MATer decomposition in streams (QUANTOMAT)

The decomposition of terrestrial organic material such as leaf litter represents a fundamental ecosystem function in streams that delivers energy for local and downstream food webs. Although agriculture dominates most regions in Europe and fungicides are applied widely, effects of currently used fungicides on the aquatic decomposer community and consequently the leaf decomposition rate are largely unknown. Also potential compensation of such hypothesised adverse effects due to nutrients or higher average water temperatures associated with climate change are not considered. Moreover, climate change is predicted to alter the community of aquatic decomposers and an open question is, whether this alteration impacts the leaf decomposition rate. The current projects follows a tripartite design to answer these research questions. Firstly, a field study in a vine growing region where fungicides are applied in large amounts will be conducted to whether there is a dose-response relationship between the exposure to fungicides and the leaf decomposition rate. Secondly, experiments in artificial streams with field communities will be carried out to assess potential compensatory mechanisms of nutrients and temperature for effects of fungicides. Thirdly, field experiments with communities exhibiting a gradient of taxa sensitive to climate change will be used to investigate potential climate-related effects on the leaf decomposition rate.

Effects of canopy structure on salinity stress in cucumber (Cucumis sativus L.)

Salinity reduces the productivity of cucumber (Cucumis sativus L.) through osmotic and ionic effects. For given atmospheric conditions we hypothesize the existence of an optimal canopy structure at which water use efficiency is maximal and salt accumulation per unit of dry matter production is minimal. This canopy structure optimum can be predicted by integrating physiological processes over the canopy using a functional-structural plant model (FSPM). This model needs to represent the influence of osmotic stress on plant morphology and stomatal conductance, the accumulation of toxic ions and their dynamics in the different compartments of the system, and their toxic effects in the leaf. Experiments will be conducted to parameterize an extended cucumber FSPM. In in-silico experiments with the FSPM we attempt to identify which canopy structure could lead to maximum long-term water use efficiency with minimum ionic stress. The results from in-silico experiments will be evaluated by comparing different canopy structures in greenhouses. Finally, the FSPM will be used to investigate to which extent the improvement of individual mechanisms of salt tolerance like reduced sensitivity of stomatal conductance or leaf expansion can contribute to whole-plant salt tolerance.

Schwerpunktprogramm (SPP) 1315: Biogeochemische Grenzflächen in Böden; Biogeochemical Interfaces in Soil, Highly-resolved imaging in artificial and natural soils to yield dynamics and structure of interfaces from oxygen, pH and water content

In soils and sediments there is a strong coupling between local biogeochemical processes and the distribution of water, electron acceptors, acids, nutrients and pollutants. Both sides are closely related and affect each other from small scale to larger scale. Soil structures such as aggregates, roots, layers, macropores and wettability differences occurring in natural soils enhance the patchiness of these distributions. At the same time the spatial distribution and temporal dynamics of these important parameters is difficult to access. By applying non-destructive measurements it is possible to overcome these limitations. Our non-invasive fluorescence imaging technique can directly quantity distribution and changes of oxygen and pH. Similarly, the water content distribution can be visualized in situ also by optical imaging, but more precisely by neutron radiography. By applying a combined approach we will clarify the formation and architecture of interfaces induces by oxygen consumption, pH changes and water distribution. We will map and model the effects of microbial and plant root respiration for restricted oxygen supply due to locally high water saturation, in natural as well as artificial soils. Further aspects will be biologically induced pH changes, influence on fate of chemicals, and oxygen delivery from trapped gas phase.

Flowering time, development and yield in oilseed rape (Brassica napus): Sequence diversity in regulatory genes

Flowering time (FTi) genes play a key role as regulators of complex gene expression networks, and the influence of these networks on other complex systems means that FTi gene expression triggers a cascade of regulatory effects with a broad global effect on plant development. Hence, allelic and expression differences in FTi genes can play a central role in phenotypic variation throughput the plant lifecycle. A prime example for this is found in Brassica napus, a phenotypically and genetically diverse species with enormous variation in vernalisation requirement and flowering traits. The species includes oilseed rape (canola), one of the most important oilseed crops worldwide. Previously we have identified QTL clusters related to plant development, seed yield and heterosis in winter oilseed rape that seem to be conserved in diverse genetic backgrounds. We suspect that these QTL are controlled by global regulatory genes that influence numerous traits at different developmental stages. Interestingly, many of the QTL clusters for yield and biomass heterosis appear to correspond to the positions of meta-QTL for FTi in spring-type and/or winter-type B. napus. Based on the hypothesis that diversity in FTi genes has a key influence on plant development and yield, the aim of this study is a detailed analysis of DNA sequence variation in regulatory FTi genes in B. napus, combined with an investigation of associations between FTi gene haplotypes, developmental traits, yield components and seed yield.

Trophic interactions in the soil of rice-rice and rice-maize cropping systems

Subproject 3 will investigate the effect of shifting from continuously flooded rice cropping to crop rotation (including non-flooded systems) and diversified crops on the soil fauna communities and associated ecosystem functions. In both flooded and non-flooded systems, functional groups with a major impact on soil functions will be identified and their response to changing management regimes as well as their re-colonization capability after crop rotation will be quantified. Soil functions corresponding to specific functional groups, i.e. biogenic structural damage of the puddle layer, water loss and nutrient leaching, will be determined by correlating soil fauna data with soil service data of SP4, SP5 and SP7 and with data collected within this subproject (SP3). In addition to the field data acquired directly at the IRRI, microcosm experiments covering the broader range of environmental conditions expected under future climate conditions will be set up to determine the compositional and functional robustness of major components of the local soil fauna. Food webs will be modeled based on the soil animal data available to gain a thorough understanding of i) the factors shaping biological communities in rice cropping systems, and ii) C- and N-flow mediated by soil communities in rice fields. Advanced statistical modeling for quantification of species - environment relationships integrating all data subsets will specify the impact of crop diversification in rice agro-ecosystems on soil biota and on the related ecosystem services.

Natural variation of flowering time due to cis-regulatory evolution of FLOWERING LOCUS T and its orthologs and paralogs in Brassica napus

In many plant species, FLOWERING LOCUS T and related proteins are the mobile signal that communicates information on photoperiod from the leaves to the shoots, where the transition to flowering is realized. FT expression is tightly controlled at the transcriptional level so that it is restricted to leaves, occurs only in appropriate photoperiods, and integrates ambient temperature and developmental cues, as well as information on biotic and abiotic stress. We previously established that FT transcription in the model plant Arabidopsis thaliana requires proximal promoter cis-elements and a distal enhancer, both evolutionary conserved among Brassicacea species. In addition, FT transcription is blocked prior vernalization in biannual accessions and vernalization-dependency of FT is controlled through a CArG-box located in the first intron that binds the transcriptional repressor FLOWERING LOCUS C (FLC). Chromatin-mediated repression by the Polycomb Group (PcG) pathway is required for photoperiod-dependent FT regulation and participates in FT expression level modulation in response to other cues.In this project, I propose to explore the available sequence data from the 1001 genome project in Arabidopsis to evaluate how often changes in regulatory cis-elements at FT have occurred and how these translate into an adaptive value. Allele-specific FT expression pattern will be measured in F1 hybrids of different accessions in response to varying environmental conditions. FT alleles that show cis-regulatory variation will be further analyzed to pinpoint the causal regulatory changes and study their effect in more detail. The allotetrapolyploid species Brassica napus is a hybrid of two Brassiceae species belonging to the A- and C-type genome, which are in turn mesopolyploid due to a genome triplication that occurred ca. 10x106 years ago. We will determine allele-specific expression of FT paralogs from both genomes of a collection of B. napus accessions. The plants will be grown in the field in changing environmental conditions to maximize the chance to detect expression variation of the paralogs. We will compare the contribution of the founder genomes to the regulation of flowering time and asses variation in this contribution. A particular focus will be to study the impact of chromatin-mediated repression on allele selection in B. napus.

Emmy Noether-Nachwuchsgruppen, Development and risk assessment of transgenic environmentally-friendly insect pest control methods for fruit flies and mosquitoes

Various species of pest insects cause substantial damage to agriculture every year, or transmit deadly diseases to animals and humans. A successful strategy to control pest insect populations is based on the Sterile Insect Technique (SIT), which uses the release of mass-reared, radiation sterilized male insects to cause infertile matings and thus reduce the pest population level. However, irradiation is not applicable to every insect species. Thus, new strategies based on genetic modifications of pest insects have been developed or are currently under investigation.The goal of the proposed research is to improve the development and ecological safety of genetically engineered (GE) insects created for enhanced biological control programs, including the SIT and new strategies based on conditional lethality. A major concern for GE insect release programs is transgene stability, and maintenance of their consistent expression. Transgene loss or intra-genomic movement could result in loss of strain attributes, and may ultimately lead to interspecies movement resulting in ecological risks. To address potential transgene instability, a new transposon vector that allows post-integration immobilization will be tested in the Mediterranean, Mexican and Oriental fruit fly tephritid pest species. In addition, the system will be established in the mosquito species Aedes and Anopheles - carriers of dengue and malaria.Random genomic insertion is also problematic for GE strain development due to genomic position effects that suppress transgene expression, and insertional mutations that negatively affect host fitness and viability. Diminished transgene expression could result in the unintended survival of conditional lethal individuals, or the inability to identify them. To target transgene vectors to defined genomic insertion sites having minimal negative effects on gene expression and host fitness, a recombinase-mediated cassette exchange (RMCE) strategy will be developed that. RMCE will also allow for stabilization of the target site, will be tested in tephritid and mosquito species, and will aid to the development of stabilized target-site strains for conditional lethal biocontrol. This will include a molecular and organismal evaluation of an RNAi-based lethality approach. Lethality based on an RNAi mechanism in the proposed insects would increase the species specificity and having multiple targets for lethality versus one target in existing systems. By seeking to improve transgene expressivity and stabilization of transposon-based vector systems, this proposal specifically addresses issues related to new GE insects by reducing their unintended spread after field release, and by limiting the possibilities for transgene introgression.

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