David Dobnik (Avtor), Ana Lazar (Avtor), Tjaša Stare (Avtor), Kristina Gruden (Avtor), Vivianne G. A. A. Vleeshouwers (Avtor), Jana Žel (Avtor)

Povzetek

Background Virus-induced gene silencing (VIGS) is an optimal tool for functional analysis of genes in plants, as the viral vector spreads throughout the plant and causes reduced expression of selected gene over the whole plant. Potato (Solanum tuberosum) is one of the most important food crops, therefore studies performing functional analysis of its genes are very important. However, the majority of potato cultivars used in laboratory experimental setups are not well amenable to available VIGS systems, thus other model plants from Solanaceae family are used (usually Nicotiana benthamiana). Wild potato relatives can be a better choice for potato model, but their potential in this field was yet not fully explored. This manuscript presents the set-up of VIGS, based on Tobacco rattle virus (TRV) in wild potato relatives for functional studies in potato–virus interactions. Results Five different potato cultivars, usually used in our lab, did not respond to silencing of phytoene desaturase (PDS) gene with TRV-based vector. Thus screening of a large set of wild potato relatives (different Solanum species and their clones) for their susceptibility to VIGS was performed by silencing PDS gene. We identified several responsive species and further tested susceptibility of these genotypes to potato virus Y (PVY) strain NTN and N. In some species we observed that the presence of empty TRV vector restricted the movement of PVY. Fluorescently tagged PVYN-GFP spread systemically in only five of tested wild potato relatives. Based on the results, Solanum venturii (VNT366-2) was selected as the most suitable system for functional analysis of genes involved in potato–PVY interaction. The system was tested by silencing two different plant immune signalling-related kinases, StWIPK and StMKK6. Silencing of StMKK6 enabled faster spreading of the virus throughout the plant, while silencing of WIPK had no effect on spreading of the virus. Conclusions The system employing S. venturii (VNT366-2) and PVYN-GFP is a suitable method for fast and simple functional analysis of genes involved in potato–PVY interactions. Additionally, a set of identified VIGS responsive species of wild potato relatives could serve as a tool for general studies of potato gene function.

Ključne besede

potato;virus-induced gene silencing;VIGS;potato virus Y;PVY;Solanum venturii;StWIPK;StMKK6;TRV;

Podatki

Jezik: Angleški jezik
Leto izida:
Tipologija: 1.01 - Izvirni znanstveni članek
Organizacija: NIB - Nacionalni inštitut za biologijo
UDK: 578
COBISS: 3877967 Povezava se bo odprla v novem oknu
ISSN: 1746-4811
Št. ogledov: 32
Št. prenosov: 8
Ocena: 0 (0 glasov)
Metapodatki: JSON JSON-RDF JSON-LD TURTLE N-TRIPLES XML RDFA MICRODATA DC-XML DC-RDF RDF

Ostali podatki

Komentar vira: Nasl. z nasl. zaslona; Opis vira z dne 30. 5. 2016;
Strani: str. 1-12
Letnik: ǂVol. ǂ12
Zvezek: ǂno. ǂ29
Čas izdaje: 2016
DOI: 10.1186/s13007-016-0129-3
ID: 24592861