doktorska disertacija
Taja Jeseničnik (Avtor), Jernej Jakše (Mentor), Nataša Štajner (Komentor)

Povzetek

RNA-interferenca (RNAi) je ohranjen mehanizem regulacije izražanja genov, prisoten v živalih, rastlinah in glivah. Ravno v kraljestvu gliv, na modelnem organizmu Neurospora crassa, je bil mehanizem RNAi intenzivno preučevan, študije pa so vodile v odkritje glavnih komponent mehanizma, ki vodijo proces utišanja, to so proteini od RNA odvisna polimeraza RNA (RdRP), Dicer (DCL) ter Argonavt (AGO). Pri filamentoznih patogenih glivah, kamor uvrščamo tudi glivo Verticillium nonalfalfae (V. nonalfalfae), ki je povzročiteljica vaskularne bolezni hmelja, je bil mehanizem RNAi izpostavljen kot eden izmed mehanizmov patogenosti gliv. Male in mikro molekulam RNA podobne RNA (milRNA) so bile pri glivah vpletene ne samo v regulacijo izražanja endogenih genov, temveč izpostavljene kot efektorji, ki jih glive transportirajo v rastlinske gostitelje, kjer regulirajo izražanje gostiteljevih obrambnih mehanizmov. Zaradi pomembnega doprinosa mehanizma RNAi k patogenosti gliv smo v doktorski disertaciji okarakterizirali mehanizem RNAi pri glivi V. nonalfalfae z identifikacijo ključnih komponent sinteze malih RNA in milRNA ter komponente utišanja tarčnih genov. Potrdili smo obstoj in izražanje dveh proteinov RdRP, dveh DCL in dveh AGO. Nadaljnje smo identificirali ter potrdili izražanje devetih milRNA glive ter potrdili aktivnost mehanizma RNAi z eksperimentalno analizo razreza tarčnih mRNA, napovedanih v anotiranem genomu glive. Z raziskavami v okviru doktorske disertacije smo prvič poročali o obstoju mehanizma RNAi pri glivi V. nonalfalfae, kar bo bistveno doprineslo k razvoju novih strategij zatiranja škodljivca ter nadzora verticilijske bolezni hmelja.

Ključne besede

RNA-interferenca;milRNA;sekvenciranje NGS;AGO;DCL;RdRP;fitopatogene glive;Verticillium nonalfalfae;hmelj;verticilijska uvelost hmelja;disertacije;

Podatki

Jezik: Slovenski jezik
Leto izida:
Tipologija: 2.08 - Doktorska disertacija
Organizacija: UL BF - Biotehniška fakulteta
Založnik: [T. Jeseničnik]
UDK: 577.2:633.791:632.4(043.3)
COBISS: 26308099 Povezava se bo odprla v novem oknu
Št. ogledov: 553
Št. prenosov: 158
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Sekundarni jezik: Angleški jezik
Sekundarni naslov: Identification and characterization of small RNA molecules in phyto-pathogenic fungus Verticillium nonalfalfae
Sekundarni povzetek: RNA interference (RNAi) is a conserved mechanism for regulating gene expression in animals, plants and fungi. The RNAi mechanism was studied extensively in the model fungus Neurospora crassa, and these studies led to the identification of the core components of the mechanism, the RNA-dependant RNA polymerase (RdRP), Dicer (DCL) and Argonaut (AGO), that mediate silencing events. In filamentous fungi, including Verticillium nonalfalfae (V. nonalfalfae), the causal agent of wilting disease of hops, the RNAi mechanism was identified as one of the pathogenic mechanisms of these fungi. Small and microRNA-like RNAs (milRNAs) were implicated not only in regulating endogenous genes but also as effectors that fungi can export into host plants, where they regulate the expression of host defence mechanisms. Due to the important contribution of RNAi to the pathogenicity of fungi, in this doctoral dissertation, we characterised the RNAi mecahanism of V. nonalfalfae by identifying the core components of small and milRNA synthesis and the components required for gene silencing. We confirmed the existence and expression of two RdRP, two DCL and two AGO proteins. Furthermore, we identified and validated the expression of nine fungal milRNAs and confirmed the activity of the RNAi by experimental analysis of the target cleavage sites of the target gene models identified in silico in the annotated fungal genome. With the studies conducted in the scope of the doctoral dissertation, we reported on the existence of RNAi in V. nonalfalfae for the first time, and the obtained knowledge will help in the development of new strategies for the management of verticillium wilt disease in hops.
Sekundarne ključne besede: RNA interference;milRNA;NGS sequencing;AGO;DCL;RdRP;pathogenic plant fungi;Verticillium nonalfalfae;hop;verticillium wilt of hops;dissertations;
Vrsta dela (COBISS): Doktorsko delo/naloga
Študijski program: 0
Konec prepovedi (OpenAIRE): 1970-01-01
Komentar na gradivo: Univ. v Ljubljani, Biotehniška fak.
Strani: XV, 154, [24] str.
ID: 11970194