Efficient virus-induced gene silencing in roots using a modified tobacco rattle virus vector
Efficient virus-induced gene silencing in roots using a modified tobacco rattle virus vector
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DOI：
10.1104/pp.104.051466
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发表时间：
2004-12-01
期刊：
影响因子：
7.4
通讯作者：
Lacomme, C
中科院分区：
文献类型：
--
作者：
Valentine, T;Shaw, J;Lacomme, C
由于它们能够引起转录后基因沉默（称为病毒诱导的基因沉默或VIGS），植物病毒越来越多地用作植物基因功能表征的反向遗传学工具。RNA病毒已显示在多种宿主植物中触发沉默，包括茄属和拟南芥属（Arabidopsis thaliana）的成员。有几个因素影响沉默反应，包括宿主范围和植物内的病毒向性。这里提出的工作表明，一个修改的烟草脆裂病毒（TRV）载体保留的辅助蛋白2b，所需的传输由一个特定的载体线虫，不仅入侵和复制广泛的整个植物，包括分生组织，但也引发了普遍的系统VIGS响应在本氏烟草，拟南芥，番茄（番茄）的根。这种持续的VIGS响应通过参与根发育（IRT 1、TTG 1 [透明种皮]、RHL 1 [无毛根]和β-微管蛋白）、侧根分生组织功能（RML 1 [根分生组织1]）和线虫抗性（Mi）的基因的沉默来例证。沉默植物的根表现出降低的目标mRNA水平和表型先前描述的突变体等位基因。TRV-2b载体显示出增加的感染性和分生组织入侵，这两者都是根组织中基因的基于VIGS的有效功能表征的关键要求。我们的数据表明，TRV辅助蛋白2b可能在控制TRV入侵的宿主调节机制中发挥重要作用。
Due to their capability of eliciting a form of posttranscriptional gene silencing (termed virus-induced gene silencing or VIGS), plant viruses are increasingly used as reverse-genetics tools for functional characterization of plant genes. RNA viruses have been shown to trigger silencing in a variety of host plants, including members of Solanacae and Arabidopsis (Arabidopsis thaliana). Several factors affect the silencing response, including host range and viral tropism within the plant. The work presented here demonstrates that a modified tobacco rattle virus (TRV) vector retaining the helper protein 2b, required for transmission by a specific vector nematode, not only invades and replicates extensively in whole plants, including meristems, but also triggers a pervasive systemic VIGS response in the roots of Nicotiana benthamiana, Arabidopsis, and tomato (Lycopersicon escylentum). This sustained VIGS response was exemplified by the silencing of genes involved in root development (IRT1, TTG1 [transparent testa glabra], RHL1 [root hairless1], and beta-tubulin), lateral root-meristem function (RML1 [root meristendess1]), and nematode resistance (Mi). Roots of silenced plants exhibit reduced levels of target mRNA and phenocopy previously described mutant alleles. The TRV-2b vector displays increased infectivity and meristem invasion, both key requirements for efficient VIGS-based functional characterization of genes in root tissues. Our data suggest that the TRV helper protein 2b may have an essential role in the host regulatory mechanisms that control TRV invasion.