中文摘要：病毒诱导的基因沉默（VIGS）是植物体内基因功能分析的最佳工具，可作为病毒载体遍布整个植物，并可引起选定的基因在整个植物中的表达减少。马铃薯是最重要的粮食作物之一，因此对马铃薯基因表现的功能分析进行研究显得尤为重要。然而，实验室实验装置中使用的大多数马铃薯品种不适合可用的VIGS系统，因此会选择使用茄科其他模式植物（通常是烟草）。本文介绍了基于野生马铃薯亲缘植物中烟草花叶病毒（TRV）的VIGS设置，可用于开展马铃薯与病毒相互作用中的功能研究。
外文摘要：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.
外文关键词：Potato；Virus-induced gene silencing；VIGS；Potato virus Y；PVY；Solanum venturii；StWIPK；StMKK6；TRV
作者：Dobnik, David；Lazar, Ana；Stare, Tjasa；等
作者单位：斯洛文尼亚国立生物学研究所
期刊名称：PLANT METHODS
期刊影响因子：3.449
出版年份：2016
出版刊次：5
点击下载：通过适用于研究马铃薯病毒诱导的基因沉默的理想模型系统Solanum venturii揭示StMKK6是植物免疫中一个重要的参与者