Boyce Thompson Institute, Ithaca, NY, USA.
Methods Mol Biol. 2022;2360:139-153. doi: 10.1007/978-1-0716-1633-8_12.
The corn leaf aphid (Rhopalosiphum maidis), a damaging pest of maize (Zea mays), is not controlled by the insecticidal proteins in commercially available transgenic crop varieties. One promising approach is to reduce aphid growth and fecundity by targeting the expression of essential genes using plant-mediated RNA interference (RNAi). Here we describe a method whereby Sugarcane Mosaic Virus (SCMV), a positive-strand RNA virus in the Potyviridae family, is used for virus-induced gene silencing (VIGS) of gene expression in R. maidis. A segment of the R. maidis target gene is cloned into SCMV, maize plants are infected with the transgenic virus, aphids are placed on the virus-infected plants and, after a few days of feeding, decreases in target gene expression and aphid reproduction are assessed. This VIGS method can be used for rapid screening of suitable RNAi targets for aphid pest control, as well as to study the in vivo function of specific aphid genes.
玉米叶蝉（Rhopalosiphum maidis）是玉米（Zea mays）的一种有害害虫，它不受商业上可用的转基因作物品种中杀虫蛋白的控制。一种有前途的方法是通过使用植物介导的 RNA 干扰（RNAi）来靶向表达必需基因，从而降低蚜虫的生长和繁殖能力。在这里，我们描述了一种方法，即利用甘蔗花叶病毒（SCMV），一种正链 RNA 病毒在马铃薯 Y 病毒科，用于玉米叶蝉基因表达的病毒诱导基因沉默（VIGS）。将玉米叶蝉的目标基因的一个片段克隆到 SCMV 中，用转基因病毒感染玉米植株，将蚜虫放在感染病毒的植株上，几天后，评估目标基因表达和蚜虫繁殖的减少情况。这种 VIGS 方法可用于快速筛选适合蚜虫防治的 RNAi 靶标，以及研究特定蚜虫基因的体内功能。
