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Identifzierung molekularer Virus-Wirt-Interaktionen in Physcomitrella patens im Verlauf von Tomato spotted wilt virus Infektionen
Antragsteller
Dr. Jan-Wolfhard Kellmann
Fachliche Zuordnung
Organismische Interaktionen, chemische Ökologie und Mikrobiome pflanzlicher Systeme
Förderung
Förderung von 2003 bis 2007
Projektkennung
Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 5415368
Novel methods for the identification of host proteins interacting with viral gene products allow new assumptions concerning basic principles of virus replication or virus spreading through infected organisms. Resultant postulations can be confirmed (or unproved) after generating of, or screening for, host mutants which are defective in steps which possibly facilitate pivotal virus functions. The moss Physcomitrella patens allows both, the analysis of mutant collections as well as, with the aid of homologous recombination, generation of a specified mutant defective in a gene of interest. To study molecular virus-plant interactions utilizing P. patens as a host, we established a system to inoculate gametophores with Tomato spotted wilt virus (TSWV). Following western as well as ELISA analysis of virus-treated gametophores, TSWV encoded structural N and non-structural NSm protein could be detected (HÜHNS ET AL., 2003 PLANT CELL TISSUE & ORGAN CULTURE, IN PRESS), assuming that P. patens supports multiplication of TSWV virions. These results encouraged us to propose further approaches ; i.) substantiation of successful P. patens infection by verifying TSWV viral messenger- and genome-complementary S, M, and L RNA species, ii.) detection of virus particles in moss cells by microscopy, combined with concurrent tests for virion infectivity (employed by re-inoculation of tobacco plants), and iii.) attempts to achieve P.patens mutants assumed to be defective in specified molecular TSWV - host interactions.
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