Flores Miguel A, Reyes Maria I, Robertson Dominique Niki, Kjemtrup Susanne
Department of Plant and Microbial Biology, North Carolina State University, Box 7612, Raleigh, NC, 27695-7612, USA.
Methods Mol Biol. 2015;1284:305-22. doi: 10.1007/978-1-4939-2444-8_15.
Coupled with the advantages afforded by the model plant Arabidopsis, virus-induced gene silencing (VIGS) offers a rapid means to assess gene function. The geminivirus vector based on Cabbage leaf curl virus described here has the benefits of small insert size and persistent silencing of the target gene through the life cycle of the plant. Here, we show that genetic variation in the vast collection of Arabidopsis accessions can be leveraged to ameliorate viral symptomology that accompanies the VIGS procedure. The plasticity of phenotypes under different day lengths or temperature conditions can be exploited to achieve maximum silencing efficacy in either vegetative or inflorescence tissue, according to the question being asked. Protocols and vectors for Agro-infiltration of primary leaves, subapical pricking in older plants, and microprojectile bombardment are described.
结合模式植物拟南芥所具有的优势，病毒诱导基因沉默（VIGS）提供了一种快速评估基因功能的方法。本文所述的基于甘蓝叶卷曲病毒的双生病毒载体具有插入片段小以及在植物生命周期中对靶基因持续沉默的优点。在这里，我们表明可以利用拟南芥大量种质资源中的遗传变异来改善VIGS过程中伴随的病毒症状。根据所提出的问题，可以利用不同日长或温度条件下的表型可塑性，在营养组织或花序组织中实现最大程度的沉默效果。文中还描述了对初生叶进行农杆菌浸润、对老龄植物进行根尖下针刺以及微粒轰击的实验方案和载体。
