Solanum venturii, a suitable model system for virus-induced gene silencing studies in potato reveals StMKK6 as an important player in plant immunity

Abstract

BackgroundVirus-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.ResultsFive 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.ConclusionsThe 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s13007-016-0129-3) contains supplementary material, which is available to authorized users.

Plant Molecular Responses to Potato Virus Y: A Continuum of Outcomes from Sensitivity and Tolerance to Resistance

“…Specifically, the MAPK signaling network in HR-conferred resistance to PVY involves MAPK kinases 6 (MKK6) and the downstream targets MAPK4_2 (orthologue of AtMAPK4, -11 and -12), MAPK6 (orthologue of AtMAPK6), and MAPK13 (orthologue of AtMAPK13) [61]. Downregulation of MKK6 was shown to increase PVY concentrations in infected plants, confirming that this kinase is an important component of potato immunity against the virus [62].…”

Characterization of Three Chalcone Synthase-like Genes in Dianthus Chinensis 

“…The Tobacco rattle virus (TRV)-based VIGS system has been developed and used for verifying gene functions in diverse plants (Chen et al 2004;Singh et al 2012;Zhong et al 2014;Dobnik et al 2016). The infection of virus vector carrying sequences of plant genes is replicated and produces double-stranded RNA molecules in infected plants.…”

“…Gaining and characterizing CHS genes involving in the anthocyanin biosynthesis pathway and the ower coloration is important to illustrate a diversity of ower colors in D. chinensis. Virus-induced gene silencing (VIGS) has been widely used in various plant species to explore the gene functions (Chen et al 2004;Quadrana et al 2011;Singh et al 2012;Manmathan et al 2013;Zhong et al 2014;Dobnik et al 2016). In this paper, three CHS genes were isolated in D. chinensis and their functions were studied using VIGS.…”

State of the Art of Genetic Engineering in Potato: From the First Report to Its Future Potential

“…Once DNA was delivered, transient or stable transformation can occur. Virus induced gene silencing is an attractive approach to generate transient loss-of-function assays to assess the role of genes in a short time, as an alternative to stable transformation ( Brigneti et al, 2004 ; Faivre-Rampant et al, 2004 ; Dobnik et al, 2016 ). In addition, A. tumefaciens -mediated infiltration without involving a viral based system has been reported to study gene function or protein localization in potato ( Bhaskar et al, 2009 ).…”