Co-Project leader: Dr. Ludmilla BorisjukEmail:
Various in vivo and topographical approaches are applied in frame of this work package, which focuses on analysis of living seed and seed performance. Developing and mature seeds will be analyzed as well as the seed germination process.
We aim to characterize the dynamics of seed biomass accumulation in three selected cultivars grown under optimal and detrimental environmental conditions (Express, AV01, and AV02). Thereby, the inner structure of the seed and tissue-specific lipid distribution will be analyzed noninvasively using NMR-based technology (Borisjuk et al., 2013a). The NMR-imaging platform (Avance III HD 400WB Spektrometer) provides high spatial resolution sufficient for quantification of individual seed organs (e.g. radicle, inner and outer cotyledons, seed coat). The quantitative 3D models will visualize seed structure and lipid topology at key developmental phases of seed development.
Histological analysis of seed will be performed - in parallel to the NMR imaging approach - using procedures well established for oilseed rape seed (Borisjuk et al. 2013b). Our histological work will efficiently determine cellular and tissue-specific changes, reflecting regular seed development and/or their aberrations.
Mature seeds are provided by NPZi (WP II.1) and include 400 lines (grown at 4 sites in years 1 and 2) and 100 lines (grown at 2 sites in year 3 and 4). Seed composition will be analysed non-invasively using TD-NMR (Rolletschek et al. 2015). In addition, hyperspectral imaging (950nm –2000nm) will be applied to individual seeds. Germinability of mature seeds will be tested using a high-throughput respiration assay (Keil et al. 2017). Detailed analysis of seeds from selected lines grown under distinct environmental conditions will allow us to make predictions on the environmental and genetic effects on germination behavior of seeds, which is one of the final aims of the project.
Output of this WP will (i) give quantitative and visual information about spatial arrangement of storage processes at the tissue/cellular level, enabling comparative analysis of seed at different developmental stages; (ii) define alteration/peculiarities of seed development in distinct cultivars in response to detrimental conditions/environmental stimuli; (iii) provide the basis for the development of an automated segmentation procedure (WP II.4) and further modeling approaches (WPs II.8/9/11).