title: Solanum venturii, a suitable model system for virus-induced gene silencing studies in potato reveals StMKK6 as an important player in plant immunity.
abstract: 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 PVY(N)-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 PVY(N)-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.
authors.affiliation: Department of Biotechnology and Systems Biology, National Institute of Biology, Večna Pot 111, 1000 Ljubljana, Slovenia.
authors.forename: David
authors.initials: D
authors.lastname: Dobnik
authors.affiliation: Department of Biotechnology and Systems Biology, National Institute of Biology, Večna Pot 111, 1000 Ljubljana, Slovenia.
authors.forename: Ana
authors.initials: A
authors.lastname: Lazar
authors.affiliation: Department of Biotechnology and Systems Biology, National Institute of Biology, Večna Pot 111, 1000 Ljubljana, Slovenia.
authors.forename: Tjaša
authors.initials: T
authors.lastname: Stare
authors.affiliation: Department of Biotechnology and Systems Biology, National Institute of Biology, Večna Pot 111, 1000 Ljubljana, Slovenia.
authors.forename: Kristina
authors.initials: K
authors.lastname: Gruden
authors.affiliation: Wageningen UR Plant Breeding, Wageningen University and Research Centre, P.O. Box 386, 6700 AJ Wageningen, The Netherlands.
authors.forename: Vivianne G A A
authors.initials: VG
authors.lastname: Vleeshouwers
authors.affiliation: Department of Biotechnology and Systems Biology, National Institute of Biology, Večna Pot 111, 1000 Ljubljana, Slovenia.
authors.forename: Jana
authors.initials: J
authors.lastname: Žel
keywords: PVY; Potato; Potato virus Y; Solanum venturii; StMKK6; StWIPK; TRV; VIGS; Virus-induced gene silencing