20.500.12556/RUL-136235 Karakterizacija efektorskega proteina Vna7.443 iz glive Verticillium nonalfalfae Characterization of effector protein Vna7.443 of Verticillium nonalfalfae Odkrivanje funkcije kandidatnih sekretornih efektorskih proteinov (CSEPs) je ključno za razumevanje interakcije med fitopatogenimi organizmi in rastlinskimi gostitelji. Hemibiotrofna fitopatogena gliva vaskularnega venenja Verticillium nonalfalfae, ki primarno okužuje hmelj (Humulus lupulus L.) sintetizira štiri opisane apoplastne efektorske proteine: VnaPRX1.1277 (peroksidaza razreda II), VnaSSP4.2, VnaUn.279 in VnaChtBP (hitin vezavni protein), znanih citoplazemskih efektorskih proteinov pa nima. Cilj magistrske naloge je bil karakterizacija kandidatnega sekretornega efektorskega proteina Vna7.443, izbranega na osnovi predhodnih študij. Proteinu Vna7.443 smo z bioinformacijskimi pristopi predvidili homologne proteine, potencialne tarčne celične procese in osnovne opisne parametre. Subcelično lokalizacijo fuzijskih proteinov Vna7.443 z mRFP (monomerni rdeči fluorescenčni protein) označbo na C- in N-terminalnem koncu smo izvedli s konfokalno mikroskopijo na listnih izsečkih Nicotiana benthamiana, ki smo jih posredno transformirali z Agrobacterium tumefaciens. Ugotovili smo, da: 1) je protein Vna7.443 najverjetneje citoplazemski efektorski protein, ki se, v primeru ko je C-terminalni konec prost, lokalizira v jedro rastlinske celice, 2) ne sproži preobčuljivostnega odziva v modelni rastlini, 3) vsebuje osem tandemskih ponovitev WD40 in 4) ima ohranjene homologe znotraj rodov gliv Verticillium, Cercospora in Fusarium. Detection of candidate secreted effector proteins (CSEPs) function is key to understanding the interaction between phytopathogenic organisms and plant hosts. The hemibiotrophic phytopathogenic vascular wilt fungus Verticillium nonalfalfae, which primarily infects hops (Humulus lupulus L.), produces four known apoplastic effector proteins VnaPRX1.1277 (class II peroxidase), VnaSSP4.2, VnaUn.279 and VnaChtBP (chitin binding protein), however, it has no known cytoplasmic effector proteins. The aim of the master's thesis was to characterize the candidate secretory effector protein Vna7.443, selected on the basis of previous studies. Homologous proteins, potential target cell processes, and basic descriptive parameters were determined for the Vna7.443 protein by bioinformatics approaches. Subcellular localization of Vna7.443 fusion proteins with mRFP labeling at the C- and N-terminal end was performed by confocal microscopy on Agrobacterium tumefaciens-transformed Nicotiana benthamiana leaf sections. The results showed that Vna7.443 protein is likely a cytoplasmic effector protein that, when the C-terminal end is free, localizes to the plant cell nucleus, but does not trigger a hypersensitive response in planta. Vna7.443 contains eight tandem WD40 repeats and has preserved homologues within fungal genuses Verticillium, Cercospora and Fusarium. fitopatogene glive hemibiotrof Verticillium nonalfalfae efektorski protein verticilijska uvelost hmelj Vna7.443 WD40 tandemske ponovitve phytopathogenic fungi hemibiotroph verticillium wilt Verticillium nonalfalfae hop effector protein Vna7.443 WD40 tandem repeats true false false [K. Dretnik] Slovenski jezik Angleški jezik Magistrsko delo/naloga 2022-04-21 07:15:43 2022-04-21 07:15:52 2022-09-15 04:13:36 0000-00-00 00:00:00 2022 0 Ljubljana 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 632.4:633.791:601.4:577.21(043.2) 203440 104982275 22732.pdf 22732.pdf 1 302F833616EC33844F27F165B1060895 c8659fceb37fe8d50d72917d3835ab48b15fa717982f1904492006bcd9c3304e 0bd1236d-c132-11ec-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=155774 Biotehniška fakulteta 0 0 0