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Research project (§ 26 & § 27)
Duration : 2022-01-01 - 2024-12-31

Cornelian cherries gain increasingly the consumer interest as healthy food, which leads to an increased market demand for high quality fruits and corresponding planting material. Previously we have analyzed more than 400 genetically unique C. mas accessions in the Pielach Valley, Traisen Valley and Gölsen Valley, both phenotypically and genetically. Findings on the long-term development of temperature and precipitation levels suggest that breeding for drought tolerance and disease resistance prevail in the face of current and future challenges with changing climatic conditions. To develop a breeding strategy for the production of novel genotypes adapted to future needs for a sustainable production in the region, the following objectives are envisaged: 1. The germination of the cornel cherry seeds takes up to 3 years and first fruits are expected in seedling plants after 8 years. Therefore, a marked reduction in the germination time would benefit the breeding, but also the propagation of Cornelian cherry. Thus, the premature breaking of the dormancy is a major issue to be worked on and improved in the present project in vivo and in vitro. 2. Valuable cultivars have to be propagated vegetatively. Grafting experiments will be carried out at different times of the year and with different scions in the greenhouse and in the insect proof screenhouse. In addition, the technique of "in vitro grafting" will be established for Cornus mas. 3. The creation of a reference genome sequence of a Pielachtal selection of Cornus mas will make it possible to identify essential genome segments including regulatory elements and provide the basis for a marker-assisted selection (MAS). 4. A MAS strategy will be developed and used on selected genotypes in the Austrian Cornelian cherry cultivation. This will allow to shorten the time until a decision is made about the breeding value of a new breed, i.e. the later growth and production behavior of a plant, as early as the seedling stage. 5. Cornelian cherries are also attacked by viruses and phytoplasmas. However, since they are mainly considered wild plants, infections are not reported, because there exist no systematic surveys. Since virus transmission via seeds cannot be ruled out in Cornelian cherry, in the present project, the testing of seedlings and planting material by ELISA or PCR will be established and a control strategy developed.
Research project (§ 26 & § 27)
Duration : 2021-09-01 - 2023-08-31

WIDER RESEARCH CONTEXT Infection numbers and death tolls of COVID-19 are still climbing globally, while countries are torn between implementing more lockdown measures and tentatively reopening some aspects of economic and public life.The common ground is that more and especially focused testing for SARS-CoV-2 is essential for preventing the uncontrolled spread of new SARS-CoV-2 variants. Real-time polymerase chain reaction (RT-PCR) from respiratory specimens is the current gold-standard for diagnosis and mutation detection, but does not discriminate between infectious virus and non-infectious traces of viral RNA. Viral growth in cell culture is a direct measure of viral infectivity of a specimen, but is not amenable for routine diagnosis owing to long turnaround times and the necessity of high biosafety containment laboratories. A rapid test that determines the SARS-CoV-2 infectivity status of patients would provide front-line healthcare workers with an urgently needed tool in clinical patient management and could help to curb the spread of COVID-19, while saving precious health system resources and avoiding unnecessarily long quarantine for patients. We aim to address this bottleneck of contemporary COVID-19 diagnosis with the development of an innovative point-of-care (POC) test that allows to rapidly determine the SARS-CoV-2 variant and infectivity status of a patient. SCIENTIFIC OBJECTIVES Production of soluble trimeric SARS-CoV-2 spike protein (S), receptor binding domain (RBD) and soluble ACE-2 receptor as well as ACE-2 mutants with superior binding affinity for S. Production of recombinant baculoviruses and HIV-1 Gag virus-like particles pseudo-typed with the SARS-CoV-2 spike surface glycoprotein for being used as test analytes. Discrimination of wildtype SARS-CoV-2 and currently-threatening mutants (i.e. B1.1.7,B.1.351, P1, etc.) on basis of differential DNA-RNA (from S-, N-, E- regions) hybridization using clinical patient samples. Understanding bio-surface adsorption mechanisms. APPROACH Our consortium joins forces and combines the complementary expertise of three partners, covering 1) innovative electronic biosensor design and construction on basis of already implemented systems detecting bio-molecules in real-time (AIT), 2) the recombinant expression of complex SARS CoV-2 antigen analytes and receptors and their biochemical analysis (BOKU) and the 3) validation of SARS-CoV-2 diagnostic tests and access to clinical patient samples (MUV). LEVEL OF INNOVATION The innovation of the project is the development of a novel and rapid electronic diagnostic POC test that allows to determine whether SARS-CoV-2 in a patient sample is still intact and infective. We achieve this by concomitant detection of virus genetic material and intact virions based on an innovative bio-sensor surface in an electronic device, referenced by an optical measurement tool. PRIMARY RESEARCHERS INVOLVED: Robert Strassl, Anna Nele Herdina, Miriam Klausberger, Patrik Aspermair
Research project (§ 26 & § 27)
Duration : 2021-12-01 - 2023-11-30

WIDER RESEARCH CONTEXT Virus–receptor interactions are pivotal in establishing an infection. Viruses, particularly RNA viruses, such as SARS-CoV-2, constantly evolve and any mutations in the viral spike surface glycoprotein, particularly in the receptor-binding domain, need to be thoroughly monitored. Also, the port of entry for SARS-CoV-2, the receptor ACE-2 displays heterogeneity among humans. The viral spike protein and the human receptor are heavily glycosylated and glycans indirectly support or are directly involved in the interaction of the two proteins. Any mutations in the viral spike protein or ACE-2 single nucleotide polymorphisms that result in the loss of a strategically-positioned glycan in or close to the binding interface therefore require our attention, as these may in the worst case increase susceptibility to viral infection and transmissibility. A profound understanding of viral–receptor interactions is of significant importance, allowing us to expand our knowledge of tissue and species tropism, pathogenesis in certain human populations and early preparedness for emerging variants of concerns. SCIENTIFIC OBJECTIVES The overall aim of this project is to produce and analyze different spike and ACE-2 variants with site-specific ablations of glycosylation sites within the binding interface, with a focus on naturally occurring ACE-2 glycovariant polymorphisms. We will evaluate the capability of the panel of interaction partners to bind under dynamic conditions and quantify binding strengths and kinetics, as well as map their interaction energy landscape. In addition, we will monitor the structural role of the spike glycomutations and their involvement in modulating its conformational dynamics and the inhibitory effect of soluble ACE-2 glycovariants on the binding of spike glycomutants to cellular ACE-2 receptors. APPROACH Soluble SARS-CoV-2 spike and ACE-2 glycomutants will be recombinantly expressed in human cell lines and extensively characterized by quantitative glycopeptide analysis. The interactions between them will be analysed in single molecule and cell force spectroscopy experiments that, unlike ensemble methods, can capture every individual binding-unbinding events. High speed AFM movies will directly film dynamic conformational changes of isolated SARS-CoV-2 spike protein glycomutants. LEVEL OF INNOVATION In contrast to previous studies using already emerged spike variants and the wildtype receptor ACE-2, we put a special focus on specific glycosylation sites of the interaction partners, that, owing to their proximal/strategic position, are suggested to affect viral-host interaction. Our comprehensive investigations will not only result in a valuable collection of data for deciphering the mechanisms of spike-ACE-2 variant interaction, but also provide an experimental basis for the design of novel therapeutics for effective blocking of viral variant entry. PI/co-PI: Peter Hinterdorfer, Miriam Klausberger

Supervised Theses and Dissertations