Faivre-Rampant Odile, Gilroy Eleanor M, Hrubikova Katarina, Hein Ingo, Millam Steve, Loake Gary J, Birch Paul, Taylor Mark, Lacomme Christophe
Programmes of Cell-to-Cell Communication, Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, United Kingdom.
Plant Physiol. 2004 Apr;134(4):1308-16. doi: 10.1104/pp.103.037507.
Virus induced gene silencing (VIGS) is increasingly used to generate transient loss-of-function assays and has potential as a powerful reverse-genetics tool in functional genomic programs as a more rapid alternative to stable transformation. A previously described potato virus X (PVX) VIGS vector has been shown to trigger silencing in the permissive host Nicotiana benthamiana. This paper demonstrates that a PVX-based VIGS vector is also effective in triggering a VIGS response in both diploid and cultivated tetraploid Solanum species. We show that systemic silencing of a phytoene desaturase gene is observed and maintained throughout the foliar tissues of potato plants and was also observed in tubers. Here we report that VIGS can be triggered and sustained on in vitro micropropagated tetraploid potato for several cycles and on in vitro generated microtubers. This approach will facilitate large-scale functional analysis of potato expressed sequence tags and provide a noninvasive reverse-genetic approach to study mechanisms involved in tuber and microtuber development.
病毒诱导的基因沉默（VIGS）越来越多地用于产生瞬时功能丧失分析，并且作为功能基因组计划中一种强大的反向遗传学工具具有潜力，是稳定转化的一种更快的替代方法。先前描述的马铃薯X病毒（PVX）VIGS载体已被证明能在允许宿主本氏烟草中引发沉默。本文证明基于PVX的VIGS载体在触发二倍体和栽培四倍体茄属物种的VIGS反应方面也有效。我们表明，在马铃薯植株的整个叶组织中观察到并维持了八氢番茄红素去饱和酶基因的系统性沉默，在块茎中也观察到了这种沉默。在这里我们报告，VIGS可以在体外微繁殖的四倍体马铃薯上触发并持续几个周期，并且在体外产生的微型块茎上也能触发并持续。这种方法将有助于对马铃薯表达序列标签进行大规模功能分析，并提供一种非侵入性的反向遗传学方法来研究块茎和微型块茎发育所涉及的机制。
