Virus induced gene silencing (VIGS) for functional analysis of wheat genes involved in Zymoseptoria tritici susceptibility and resistance.
Virus induced gene silencing (VIGS) for functional analysis of wheat genes involved in Zymoseptoria tritici susceptibility and resistance.
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DOI：
10.1016/j.fgb.2015.04.006
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发表时间：
2015-06
期刊：
影响因子：
--
通讯作者：
Kanyuka K
中科院分区：
文献类型：
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作者：
Lee WS;Rudd JJ;Kanyuka K
病毒诱导的基因沉默(VIGS)是研究基因功能的有力工具。将大麦条纹花叶病毒(BSMV)载体用于小麦VIGS。我们采用BSMV和VIGS来研究小麦与小麦酵母的相互作用。在这里，我们介绍我们的详细和最新的BSMV VIGS协议。病毒诱导的基因沉默(VIGS)在植物中已经成为一种强大的反向遗传技术，可以补充稳定的转基因RNAi，在某些物种中，VIGS已经成为一种可行的基因功能分析的替代方法。大麦条纹花叶病毒(BSMV)是21世纪初作为VIGS载体发展起来的一种核糖核酸病毒，并已被用于研究小麦基因的功能。BSMV载体有几种变体，其中一些需要在体外转录具有感染性的病毒RNA，而另一些则依赖于通过基因枪法或根癌农杆菌将基于DNA的载体输送到植物细胞中生产病毒RNA。我们采用了最新一代的BSMV VIGS载体来鉴定和研究与小麦ZymosepVictoria tritici相互作用的相关基因，并在这里介绍了详细和最新的协议。
Virus-induced gene silencing (VIGS) is a powerful tool for studying gene function. Barley stripe mosaic virus (BSMV) vector is used for VIGS in wheat. We adapted BSMV VIGS to study wheat–Zymoseptoria tritici interactions. Here we present our detailed and the most up-to-date protocols for BSMV VIGS. Virus-induced gene silencing (VIGS) has emerged as a powerful reverse genetic technology in plants supplementary to stable transgenic RNAi and, in certain species, as a viable alternative approach for gene functional analysis. The RNA virus Barley stripe mosaic virus (BSMV) was developed as a VIGS vector in the early 2000s and since then it has been used to study the function of wheat genes. Several variants of BSMV vectors are available, with some requiring in vitro transcription of infectious viral RNA, while others rely on in planta production of viral RNA from DNA-based vectors delivered to plant cells either by particle bombardment or Agrobacterium tumefaciens. We adapted the latest generation of binary BSMV VIGS vectors for the identification and study of wheat genes of interest involved in interactions with Zymoseptoria tritici and here present detailed and the most up-to-date protocols.