Institute of Sugar Beet Research, Göttingen, Germany.
Institute of Horticultural Production Systems, Leibniz University Hannover, Hannover, Germany.
Mol Plant Pathol. 2023 Oct;24(10):1319-1329. doi: 10.1111/mpp.13372. Epub 2023 Jul 6.
In the field of plant virology, the usage of reverse genetic systems has been reported for multiple purposes. One is understanding virus-host interaction by labelling viral cDNA clones with fluorescent protein genes to allow visual virus tracking throughout a plant, albeit this visualization depends on technical devices. Here we report the first construction of an infectious cDNA full-length clone of beet mosaic virus (BtMV) that can be efficiently used for Agrobacterium-mediated leaf inoculation with high infection rate in Beta vulgaris, being indistinguishable from the natural virus isolate regarding symptom development and vector transmission. Furthermore, the BtMV clone was tagged with the genes for the monomeric red fluorescent protein or the Beta vulgaris BvMYB1 transcription factor, which activates the betalain biosynthesis pathway. The heterologous expression of BvMYB1 results in activation of betalain biosynthesis genes in planta, allowing visualization of the systemic BtMV spread with the naked eye as red pigmentation emerging throughout beet leaves. In the case of BtMV, the BvMYB1 marker system is stable over multiple mechanical host passages, allows qualitative as well as quantitative virus detection and offers an excellent opportunity to label viruses in plants of the order Caryophyllales, allowing an in-depth investigation of virus-host interactions on the whole plant level.
在植物病毒学领域,已经有报道称,反向遗传系统被用于多种目的。其中一个目的是通过标记带有荧光蛋白基因的病毒 cDNA 克隆来了解病毒-宿主相互作用,从而可以在整个植物中对病毒进行可视化追踪,尽管这种可视化依赖于技术设备。在这里,我们首次构建了甜菜曲顶病毒(BtMV)的全长感染性 cDNA 克隆,该克隆可通过农杆菌介导的叶片接种高效使用,在感染率方面与天然病毒分离株相似,在症状发育和载体传播方面没有区别。此外,BtMV 克隆被标记了单体红色荧光蛋白或甜菜 Beta vulgaris BvMYB1 转录因子的基因,该转录因子激活甜菜红素生物合成途径。BvMYB1 的异源表达导致植物体内甜菜红素生物合成基因的激活,允许用肉眼观察到 BtMV 的系统传播,因为红色素会在整个甜菜叶片中出现。在 BtMV 的情况下,BvMYB1 标记系统在多次机械传代过程中稳定,允许进行定性和定量的病毒检测,并为标记 Caryophyllales 目中的病毒提供了极好的机会,从而可以深入研究整个植物水平上的病毒-宿主相互作用。