Department of Plant Pathology, University of California, Davis, 95616 CA, USA.
Viruses. 2018 Apr 24;10(5):216. doi: 10.3390/v10050216.
Plant virus-based vectors are valuable tools for recombinant gene expression and functional genomics for both basic and applied research. In this study, (LIYV) of the genus was engineered into a virus vector that is applicable for efficient protein expression and virus-induced gene silencing (VIGS) in plants. We examined gene replacement and “add a gene” strategies to develop LIYV-derived vectors for transient expression of the green fluorescent protein (GFP) reporter in plants. The latter yielded higher GFP expression and was further examined by testing the effects of heterologous controller elements (CEs). A series of five vector constructs with progressively extended LIYV CP sgRNA CEs were tested, the longest CE gave the highest GFP expression but lower virus accumulation. The whitefly transmissibility of the optimized vector construct to other host plants, and the capability to accommodate and express a larger gene, a 1.8 kb β-glucuronidase (GUS) gene, were confirmed. Furthermore, the LIYV vector was also validated VIGS by silencing the endogenous gene, (PDS) in plants, and the transgene GFP in line 16c plants. Therefore, LIYV-derived vectors could provide a technical reference for developing vectors of other economically important criniviruses.
基于植物病毒的载体是用于重组基因表达和功能基因组学的有价值的工具，无论是基础研究还是应用研究。在这项研究中，将属的 (LIYV) 工程化为一种病毒载体，可在植物中有效表达蛋白质和诱导病毒基因沉默 (VIGS)。我们研究了基因替换和“添加基因”策略，以开发用于瞬时表达绿色荧光蛋白 (GFP) 报告基因的 LIYV 衍生载体。后者产生了更高的 GFP 表达，并通过测试异源控制器元件 (CE) 的效果进一步进行了检验。我们测试了一系列带有逐渐延长的 LIYV CP sgRNA CE 的五个载体构建体，最长的 CE 给出了最高的 GFP 表达，但病毒积累较低。已优化的载体构建体对其他宿主植物的粉虱传播能力以及容纳和表达更大基因（1.8 kb β-葡萄糖醛酸酶 (GUS) 基因）的能力得到了确认。此外，LIYV 载体还通过沉默内源基因（在 植物中的 PDS）和转基因 GFP 在 16c 植物中的表达验证了 VIGS。因此，LIYV 衍生的载体可为开发其他经济重要的曲叶病毒载体提供技术参考。
