Bunwaree Heemee Devi, Klein Elodie, Saubeau Guillaume, Desprez Bruno, Ziegler-Graff Véronique, Gilmer David
Institut de Biologie Moléculaire des Plantes, CNRS-UPR 2357, Université de Strasbourg, 67000 Strasbourg, France.
Florimond Desprez, 3 rue Florimond Desprez, 59242 Cappelle-en-Pévèle, France.
Viruses. 2024 Nov 23;16(12):1823. doi: 10.3390/v16121823.
Since the ban of neonicotinoid insecticides in the European Union, sugar beet production is threatened by outbreaks of virus yellows (VY) disease, caused by several aphid-transmitted viruses, including the polerovirus beet mild yellowing virus (BMYV). As the symptoms induced may vary depending on multiple infections and other stresses, there is an urgent need for fast screening tests to evaluate resistance/tolerance traits in sugar beet accessions. To address this issue, we exploited the virus-induced gene silencing (VIGS) system, by introducing a fragment of a gene involved in chlorophyll synthesis in the BMYV genome. This recombinant virus was able to generate early clear vein chlorosis symptoms in infected sugar beets, allowing easy and rapid visual discernment of infected plants across five sugar beet lines. The recombinant virus displayed similar infectivity as the wild-type, and the insert remained stable within the viral progeny. We demonstrated that the percentage of VIGS-symptomatic plants was representative of the infection rate of each evaluated line, and depending on the susceptibility of the line to BMYV infection, VIGS symptoms may last over months. Our work provides a polerovirus-based VIGS system adapted to sugar beet crop allowing visual and rapid large-scale screens for resistance or functional genomic studies.
自欧盟禁止使用新烟碱类杀虫剂以来,甜菜生产受到病毒黄化(VY)病爆发的威胁,该病由多种蚜虫传播的病毒引起,包括马铃薯卷叶病毒属的甜菜轻度黄化病毒(BMYV)。由于诱导的症状可能因多种感染和其他胁迫而有所不同,因此迫切需要快速筛选试验来评估甜菜种质的抗性/耐受性特征。为了解决这个问题,我们利用病毒诱导基因沉默(VIGS)系统,将一个参与叶绿素合成的基因片段引入BMYV基因组。这种重组病毒能够在受感染的甜菜中产生早期明显的叶脉黄化症状,从而能够轻松快速地在五个甜菜品系中直观辨别受感染的植株。重组病毒表现出与野生型相似的感染力,并且插入片段在病毒后代中保持稳定。我们证明,出现VIGS症状的植株百分比代表了每个评估品系的感染率,并且根据品系对BMYV感染的敏感性,VIGS症状可能会持续数月。我们的工作提供了一种适用于甜菜作物的基于马铃薯卷叶病毒属的VIGS系统,可用于视觉和快速大规模筛选抗性或进行功能基因组学研究。