Brigneti Gianinna, Martín-Hernández Ana M, Jin Hailing, Chen Judy, Baulcombe David C, Baker Barbara, Jones Jonathan D G
The Sainsbury Laboratory, John Innes Centre, Norwich Research Park, Colney Lane, Norwich NR4 7UH, UK.
Plant J. 2004 Jul;39(2):264-72. doi: 10.1111/j.1365-313X.2004.02122.x.
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.
病毒诱导的基因沉默（VIGS）已在本氏烟草中常规用于评估候选基因的功能，并作为发现不同途径（尤其是抗病信号传导）所需新基因的一种方法。最近研究表明VIGS在拟南芥和番茄中也有效。在此，我们报告使用烟草脆裂病毒（TRV）病毒载体的VIGS可用于多种茄属植物，不过载体的选择和实验条件会因所研究的物种而异。我们已成功在二倍体野生种马铃薯和冈田氏茄、栽培四倍体马铃薯以及远缘六倍体亲缘种龙葵（俗称颠茄）中沉默了八氢番茄红素去饱和酶（PDS）基因。为了测试该系统是否可作为一种快速评估马铃薯及其野生近缘种中候选抗性（R）基因功能的方法，我们在马铃薯中沉默了R1和Rx，在马铃薯中沉默了RB。在离体叶片试验中，R1、Rx和RB的沉默成功减弱了R基因介导的抗病性，并导致了感病表型。因此，VIGS系统是快速评估马铃薯基因功能的有效方法。
