Kang Bomi, Venkatesh Jelli, Lee Joung-Ho, Kim Jung-Min, Kwon Jin-Kyung, Kang Byoung-Cheorl
Interdisciplinary Program in Agricultural Biotechnology, College of Agriculture and Life Sciences, Seoul National University, Seoul, 08826, Republic of Korea.
Department of Agriculture, Forestry and Bioresources, Research Institute of Agriculture and Life Sciences, Plant Genomics Breeding Institute, College of Agriculture and Life Sciences, Seoul National University, Seoul, 08826, Republic of Korea.
Plant Cell Rep. 2025 Feb 22;44(3):62. doi: 10.1007/s00299-025-03440-x.
Tobacco etch virus accumulation declined in Nicotiana benthamiana eEF1Bγ gene-edited lines, suggesting that eEF1Bγ may be a host factor for this virus. Viruses use host factors to replicate and move from cell to cell. Therefore, the editing of genes encoding viral host factors that are not essential for plant survival enables the rapid development of plants with durable virus resistance. Eukaryotic initiation factors, such as eIF4E and eIF4G, function as host factors for viral infection, and loss-of-function mutations of these factors lead to virus resistance. Broadening the spectrum of host factor targets would help expand resources for engineering virus resistance. In this study, we tested whether editing the eukaryotic translation elongation factor gene eEF1Bγ would produce virus-resistant plants. Accordingly, we targeted the four eEF1Bγ genes in Nicotiana benthamiana for editing using virus-induced gene editing (VIGE) with Tobacco rattle virus (TRV). Although we attempted to obtain plants edited for all four eEF1Bγ homologs, we failed to identify such plants. Instead, we obtained plants with three of the four homologs knocked out, harboring 1-bp insertion/deletions resulting in premature stop codons. These eEF1Bγ-edited plants did not exhibit resistance to Potato virus X (PVX), Tobacco mosaic virus (TMV), or Tomato bushy stunt virus (TBSV) but showed reduced accumulation of Tobacco etch virus (TEV) compared to wild-type plants. These findings demonstrate the feasibility of conferring resistance in plants through gene editing of eEF1Bγ, underscoring the importance of exploring diverse host factor targets for comprehensive virus resistance.
烟草蚀刻病毒在本氏烟草eEF1Bγ基因编辑品系中的积累减少，这表明eEF1Bγ可能是该病毒的一个宿主因子。病毒利用宿主因子进行复制并在细胞间移动。因此，对植物生存非必需的编码病毒宿主因子的基因进行编辑，能够快速培育出具有持久病毒抗性的植物。真核生物起始因子，如eIF4E和eIF4G，作为病毒感染的宿主因子发挥作用，这些因子的功能丧失突变会导致病毒抗性。拓宽宿主因子靶点的范围将有助于扩大工程化抗病毒的资源。在本研究中，我们测试了编辑真核生物翻译延伸因子基因eEF1Bγ是否会产生抗病毒植物。因此，我们使用烟草脆裂病毒（TRV）介导的病毒诱导基因编辑（VIGE）技术，靶向编辑本氏烟草中的四个eEF1Bγ基因。尽管我们试图获得所有四个eEF1Bγ同源基因都被编辑的植物，但未能鉴定出此类植物。相反，我们获得了四个同源基因中有三个被敲除的植物，这些植物含有1个碱基的插入/缺失，导致提前出现终止密码子。这些eEF1Bγ编辑的植物对马铃薯X病毒（PVX）、烟草花叶病毒（TMV）或番茄丛生矮缩病毒（TBSV）均无抗性，但与野生型植物相比，烟草蚀刻病毒（TEV）的积累有所减少。这些发现证明了通过对eEF1Bγ进行基因编辑赋予植物抗性的可行性，强调了探索多种宿主因子靶点以实现全面抗病毒的重要性。
