Muruganantham Mookkan, Moskovitz Yoni, Haviv Sabrina, Horesh Tamar, Fenigstein Annie, Preez Jacques du, Stephan Dirk, Burger Johan T, Mawassi Munir
The S. Tolkowsky Laboratory, Department of Plant Pathology-The Virology Unit, Agricultural Research Organization, Volcani Center, Bet Dagan 50250, Israel.
J Virol Methods. 2009 Feb;155(2):167-74. doi: 10.1016/j.jviromet.2008.10.010. Epub 2008 Dec 2.
Virus-induced gene silencing (VIGS) is an attractive approach for studying gene function. Although the number of virus vectors available for use in VIGS experiments has increased in recent years, most of these vectors are applied in annual or herbaceous plants. The aim of this work was to develop a VIGS vector based on the Grapevine virus A (GVA), which is a member of the genus Vitivirus, family Flexiviridae. The GVA vector was used to silence the endogenous phytoene desaturase (PDS) gene in Nicotiana benthamiana plants. In addition, an Agrobacterium-mediated method for inoculating micropropagated Vitis vinifera cv. Prime plantlets via their roots was developed. Using this method, it was possible to silence the endogenous PDS gene in V. vinifera plantlets. The GVA-derived VIGS vector may constitute an important tool for improving functional genomics in V. vinifera.
病毒诱导的基因沉默(VIGS)是研究基因功能的一种有吸引力的方法。尽管近年来可用于VIGS实验的病毒载体数量有所增加,但这些载体大多应用于一年生或草本植物。这项工作的目的是开发一种基于葡萄病毒A(GVA)的VIGS载体,GVA是柔线病毒科葡萄病毒属的成员。GVA载体用于沉默本氏烟草植株中的内源性八氢番茄红素去饱和酶(PDS)基因。此外,还开发了一种通过根接种微繁殖的酿酒葡萄品种“Prime”幼苗的农杆菌介导方法。使用这种方法,可以沉默酿酒葡萄幼苗中的内源性PDS基因。源自GVA的VIGS载体可能构成改善酿酒葡萄功能基因组学的重要工具。