345 research outputs found

    Progressive 35S promoter methylation increases rapidly during vegetative development in transgenic Nicotiana attenuata plants

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    BACKGROUND: Genetically modified plants are widely used in agriculture and increasingly in ecological research to enable the selective manipulation of plant traits in the field. Despite their broad usage, many aspects of unwanted transgene silencing throughout plant development are still poorly understood. A transgene can be epigenetically silenced by a process called RNA directed DNA methylation (RdDM), which can be seen as a heritable loss of gene expression. The spontaneous nature of transgene silencing has been widely reported, but patterns of acquirement remain still unclear. RESULTS: Transgenic wild tobacco plants (Nicotiana attenuata) expressing heterologous genes coding for antimicrobial peptides displayed an erratic and variable occurrence of transgene silencing. We focused on three independently transformed lines (PNA 1.2, PNA 10.1 and ICE 4.4) as they rapidly lost the expression of the resistance marker and down-regulated transgene expression by more than 200 fold after only one plant generation. Bisulfite sequencing indicated hypermethylation within the 35S and NOS promoters of these lines. To shed light on the progress of methylation establishment, we successively sampled leaf tissues from different stages during plant development and found a rapid increase in 35S promoter methylation during vegetative growth (up to 77% absolute increase within 45 days of growth). The levels of de novo methylation were inherited by the offspring without any visible discontinuation. A secondary callus regeneration step could interfere with the establishment of gene silencing and we found successfully restored transgene expression in the offspring of several regenerants. CONCLUSIONS: The unpredictability of the gene silencing process requires a thorough selection and early detection of unstable plant lines. De novo methylation of the transgenes was acquired solely during vegetative development and did not require a generational change for its establishment or enhancement. A secondary callus regeneration step provides a convenient way to rescue transgene expression without causing undesirable morphological effects, which is essential for experiments that use transformed plants in the analysis of ecologically important traits

    Evaluation bibliothekarischer Online-Angebote: Usability und Usefulness – zwei Seiten einer Medaille

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    DOI: 10.12685/027.7-1-3-39Aus Anwendersicht sind für die Qualität bibliothekarischer Online-Angebote zwei Aspekte entscheidend. Einerseits müssen für sie relevante Inhalte zur Verfügung stehen. Andererseits sollte es mög­lichst einfach und intuitiv möglich sein, diese Inhalte zu finden und auf sie zuzugreifen. Die Themen Usefulness und Usability sind daher für Bibliotheken von grosser Bedeutung. Der Beitrag beschreibt wie diese beiden Aspekte evaluiert werden können und wie Bibliotheken anhand von BibEval und PECI selb­ständig ihre Angebote überprüfen können.From the users’ point of view two aspects are essential for the quality of library online services. On the one hand, relevant content must be available. On the other hand, it should be simple and intuitive to find and to access this content. Therefore, the topics usability and usefulness are of great importance for li­braries. This article describes how these two aspects can be evaluated and how libraries can use BibEval and PECI for assessments of their own services

    Particle Mass Concentrations and Number Size Distributions in 40 Homes in Germany : Indoor-to-outdoor Relationships, Diurnal and Seasonal Variation

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    Few studies investigated residential particle concentration levels with a full picture of aerosol particles from 10 mu m to 10 mu m size range with size-resolved information, and none was performed in central Europe in the long-term in multiple homes. To capture representative diurnal and seasonal patterns of exposure to particles, and investigate the driving factors to their variations, measurements were performed in 40 homes for around two weeks each in Leipzig and Berlin, Germany. These over 500 days' measurements combined PM10 and PM2. 5 mass concentrations, particle number concentration and size distribution (PNC and PNSD, 10-800 nm), CO2 concentration, and residential activities diary into a unique dataset. Natural ventilation was dominated, the mean ventilation rate calculated from CO2 measurements was 0.2 h(-1) and 3.7 h(-1) with closed and opened windows, respectively. The main findings of this study showed that, the residents in German homes were exposed to a significantly higher mass concentration of coarse particles than outdoors, thus indoor exposure to coarse particles cannot be described by outdoors. The median indoor PNC diurnal cycles were generally lower than outdoors (median I/O ratio 0.69). However, indoor exposure to particles was different in the cold and warm season. In the warm season, due to longer opening window periods, indoor sources' contribution was weakened, which also resulted in the indoor PNC and PNSD being very similar to the outdoors. In the cold season, indoor sources caused strong peaks of indoor PNC that exceeded outdoors, along with the relatively low penetration factor - 0.5 for all size ranges, and indoor particle losses, which was particularly effective in reducing the ultrafine PNC, resulting in a different particle exposure load than outdoors. This study provides a detailed understanding of residential particle exposure in multiple homes, facilitating future studies to assess health effects in residential environments.Peer reviewe

    Intercomparison of 15 aerodynamic particle size spectrometers (APS 3321): uncertainties in particle sizing and number size distribution

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    Aerodynamic particle size spectrometers are a well-established method to measure number size distributions of coarse mode particles in the atmosphere. Quality assurance is essential for atmospheric observational aerosol networks to obtain comparable results with known uncertainties. In a laboratory study within the framework of ACTRIS (Aerosols, Clouds, and Trace gases Research Infrastructure Network), 15 aerodynamic particle size spectrometers (APS model 3321, TSI Inc., St. Paul, MN, USA) were compared with a focus on flow rates, particle sizing, and the unit-to-unit variability of the particle number size distribution. Flow rate deviations were relatively small (within a few percent), while the sizing accuracy was found to be within 10 % compared to polystyrene latex (PSL) reference particles. The unit-to-unit variability in terms of the particle number size distribution during this study was within 10 % to 20 % for particles in the range of 0.9 up to 3 µm, which is acceptable for atmospheric measurements. For particles smaller than that, the variability increased up to 60 %, probably caused by differences in the counting efficiencies of individual units. Number size distribution data for particles smaller than 0.9 µm in aerodynamic diameter should only be used with caution. For particles larger than 3 µm, the unit-to-unit variability increased as well. A possible reason is an insufficient sizing accuracy in combination with a steeply sloping particle number size distribution and the increasing uncertainty due to decreasing counting. Particularly this uncertainty of the particle number size distribution must be considered if higher moments of the size distribution such as the particle volume or mass are calculated, which require the conversion of the aerodynamic diameter measured to a volume equivalent diameter. In order to perform a quantitative quality assurance, a traceable reference method for the particle number concentration in the size range 0.5–3 µm is needed.JRC.H.2-Air and Climat

    Biomass burning and urban emission impacts in the Andes Cordillera region based on in situ measurements from the Chacaltaya observatory, Bolivia (5240 m a.s.l.)

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    This study documents and analyses a 4-year continuous record of aerosol optical properties measured at the Global Atmosphere Watch (GAW) station of Chacaltaya (CHC; 5240 m a.s.l.), in Bolivia. Records of particle light scattering and particle light absorption coefficients are used to investigate how the high Andean Cordillera is affected by both long-range transport and by the fast-growing agglomeration of La Paz-El Alto, located approximately 20 km away and 1.5 km below the sampling site. The extended multiyear record allows us to study the properties of aerosol particles for different air mass types, during wet and dry seasons, also covering periods when the site was affected by biomass burning in the Bolivian lowlands and the Amazon Basin. The absorption, scattering, and extinction coefficients (median annual values of 0.74, 12.14, and 12.96 Mm(-1) respectively) show a clear seasonal variation with low values during the wet season (0.57, 7.94, and 8.68 Mm(-1) respectively) and higher values during the dry season (0.80, 11.23, and 14.51 Mm(-1) respectively). The record is driven by variability at both seasonal and diurnal scales. At a diurnal scale, all records of intensive and extensive aerosol properties show a pronounced variation (daytime maximum, night-time minimum), as a result of the dynamic and convective effects. The particle light absorption, scattering, and extinction coefficients are on average 1.94, 1.49, and 1.55 times higher respectively in the turbulent thermally driven conditions than the more stable conditions, due to more efficient transport from the boundary layer. Retrieved intensive optical properties are significantly different from one season to the other, reflecting the changing aerosol emission sources of aerosol at a larger scale. Using the wavelength dependence of aerosol particle optical properties, we discriminated between contributions from natural (mainly mineral dust) and anthropogenic (mainly biomass burning and urban transport or industries) emissions according to seasons and local circulation. The main sources influencing measurements at CHC are from the urban area of La Paz-El Alto in the Altiplano and from regional biomass burning in the Amazon Basin. Results show a 28 % to 80 % increase in the extinction coefficients during the biomass burning season with respect to the dry season, which is observed in both tropospheric dynamic conditions. From this analysis, long-term observations at CHC provide the first direct evidence of the impact of biomass burning emissions of the Amazon Basin and urban emissions from the La Paz area on atmospheric optical properties at a remote site all the way to the free troposphere.Peer reviewe
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