Virus-induced gene silencing in Solanum species
Virus-induced gene silencing in Solanum species
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DOI：
10.1111/j.1365-313x.2004.02122.x
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发表时间：
2004-07-01
期刊：
影响因子：
7.2
通讯作者：
Jones, JDG
中科院分区：
文献类型：
--
作者：
Brigneti, G;Martín-Hernández, AM;Jones, JDG
病毒诱导的基因沉默（VIGS）已被常规用于benthamiana中，以评估候选基因的功能，并作为发现不同途径（特别是抗病信号）所需的新基因的一种方法。VIGS最近被证明在拟南芥和番茄中起作用。本文报道了烟草摇铃病毒（TRV）病毒载体的VIGS可用于几种茄属植物，尽管载体的选择和实验条件因所研究的物种而异。我们成功地沉默了二倍体野生种龙葵（Solanum bulbocastanum）和秋葵（S. okadae）、栽培的四倍体龙葵（S. tuberosum）和遥远的六倍体近亲龙葵（S. nigrum，俗称致命龙葵）的植物烯去饱和酶（phytoene desaturase， PDS）基因。为了检验该系统是否可以作为快速评估马铃薯及其野生近缘种候选抗性基因(R)基因功能的方法，我们沉默了马铃薯抗性候选基因R1和Rx，沉默了马铃薯抗性候选基因RB。R1、Rx和RB的沉默成功地减弱了r基因介导的抗病性，并在离体叶片试验中产生了易感表型。因此，VIGS系统是马铃薯基因功能快速鉴定的有效方法。
Virus-induced gene silencing (VIGS) has been used routinely in Nicotiana benthamiana to assess functions of candidate genes and as a way to discover new genes required for diverse pathways, especially disease resistance signalling. VIGS has recently been shown to work in Arabidopsis thaliana and in tomato. Here, we report that VIGS using the tobacco rattle virus (TRV) viral vector can be used in several Solanum species, although the choice of vector and experimental conditions vary depending on the species under study. We have successfully silenced the phytoene desaturase (PDS) gene in the diploid wild species Solanum bulbocastanum and S. okadae, in the cultivated tetraploid S. tuberosum and in the distant hexaploid relative S. nigrum (commonly known as deadly nightshade). To test whether the system could be utilised as a rapid way to assess gene function of candidate resistance (R) genes in potato and its wild relatives, we silenced R1 and Rx in S. tuberosum and RB in S. bulbocastanum. Silencing of R1, Rx and RB successfully attenuated R-gene-mediated disease resistance and resulted in susceptible phenotypes in detached leaf assays. Thus, the VIGS system is an effective method of rapidly assessing gene function in potato.