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Funkcijska analiza kandidatnih efektorskih proteinov glive Verticillium nonalfalfae
Volk, Helena (Author), Javornik, Branka (Mentor) More about this mentor... This link opens in a new window

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Abstract
Pri hmelju (Humulus lupulus L.) je poznana letalna oblika verticilijske uvelosti, ki se odraža z bolezenskimi znaki uvelosti, klorozami, nekrozami in končnim propadom rastline. Bolezen povzroča fitopatogena gliva Verticillium nonalfalfae (Vna), ki za uspešno kolonizacijo rastline izloča efektorske proteine. Ti jo ščitijo pred obrambnim odzivom rastline, modulirajo imunski odziv ali vplivajo na fiziologijo rastline. Razumevanje funkcije kandidatnih sekretornih efektorskih proteinov (CSEPs) in poznavanje molekul gostitelja, s katerimi določen CSEP interagira, je ključno pri načrtovanju novih strategij zatiranja verticiljske uvelosti in lahko pospeši žlahtnjenje kultivarjev hmelja odpornih na Vna. V okviru te doktorske disertacije smo okarakterizirali kandidatna efektorska proteina VnaChtBP in VnaSSP4.2, ki smo ju izbrali na osnovi predhodnih študij. S konfokalno mikroskopijo smo pri modelni rastlini Nicotiana benthamiana ugotovili nukleo-citosolno subcelično lokalizacijo obeh izbranih CSEPs. Z infiltracijo rekombinantnih proteinov VnaChtBP in VnaSSP4.2 v različne rastline nismo uspeli potrditi hipoteze o njihovi vpletenosti v PTI ali ETI odziv. Ugotovili smo, da se efektor VnaChtBP specifično veže na hitin in ščiti glivo pred razgradnjo z rastlinskimi hitinazami. Pokazali smo, da se rekombinantni protein VnaSSP4.2 veže na molekule PIP, kardiolipin in sulfatid, ki so opisani kot modulatorji imunskega odziva.

Language:Slovenian
Keywords:fitopatogene glive, Verticillium nonalfalfae, efektor, efektorji, efektorski proteini, verticilijska uvelost, hmelj, VnaChtBP, VnaSSP4.2, hitin, lipidi
Work type:Doctoral dissertation (mb31)
Tipology:2.08 - Doctoral Dissertation
Organization:BF - Biotechnical Faculty
Year:2019
COBISS.SI-ID:933239 This link opens in a new window
Views:670
Downloads:227
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Secondary language

Language:English
Title:Functional analysis of Verticillium nonalfalfae candidate effector proteins
Abstract:
Outbreaks of a lethal type of Verticillium wilt of hops (Humulus lupulus L.) are reported yearly. This disease is caused by a phytopathogenic fungus Verticillium nonalfalfae (Vna) and presents in wilting, chlorosis, necrosis, and plant death. In order to successfully colonize its host, Vna secrets effector proteins, which protect the fungus and modulate the plant immune system or plant physiology. Understanding the function of candidate secreted effector proteins (CSEPs) and knowledge of host interacting molecules is a crucial contribution to development of effective strategies for pest control and can accelerate breeding of Vna resistant cultivars. In the presented doctoral thesis, we characterized candidate effector proteins VnaChtBP and VnaSSP4.2, which were selected based on previous studies. Using confocal microscopy, a nucleo-cytosolic subcellular localization was determined for both effectors expressed in model plant Nicotiana benthamiana. Infiltration of recombinant proteins to various plants did not confirm the hypothesis of VnaChtBP or VnaSSP4.2 involvement in PTI or ETI responses. We were able to show that the recombinant protein VnaSSP4.2 binds to PIP, cadiolipin and sulfatide molecules, all known modulators of plant immunity. We determined that the effector VnaChtBP binds specifically to chitin and protects the fungus from degradation by plant chitinases.

Keywords:phytopathogenic fungi, Verticillium nonalfalfae, effector, effectors, effector proteins, Verticillium wilt, hops, VnaChtBP, VnaSSP4.2, chitin, lipids

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