Mardini Majd, Kazancev Mikhail, Ivoilova Elina, Utkina Victoria, Vlasova Anastasia, Demurin Yakov, Soloviev Alexander, Kirov Ilya
All-Russia Research Institute of Agricultural Biotechnology, Timiryazevskaya Str. 42, 127550, Moscow, Russia.
Moscow Institute of Physics and Technology, 141701, Dolgoprudny, Russia.
Plant Methods. 2024 Aug 12;20(1):122. doi: 10.1186/s13007-024-01241-z.
Virus-Induced Gene Silencing (VIGS) is a versatile tool in plant science, yet its application to non-model species like sunflower demands extensive optimization due to transformation challenges. In this study, we aimed to elucidate the factors that significantly affect the efficiency of Agrobacterium-VIGS in sunflowers. After testing a number of approaches, we concluded that the seed vacuum technique followed by 6 h of co-cultivation produced the most efficient VIGS results. Genotype-dependency analysis revealed varying infection percentages (62-91%) and silencing symptom spreading in different sunflower genotypes. Additionally, we explored the mobility of tobacco rattle virus (TRV) and phenotypic silencing manifestation (photo-bleaching) across different tissues and regions of VIGS-infected sunflower plants. We showed the presence of TRV is not necessarily limited to tissues with observable silencing events. Finally, time-lapse observation demonstrated a more active spreading of the photo-bleached spots in young tissues compared to mature ones. This study not only offers a robust VIGS protocol for sunflowers but also provides valuable insights into genotype-dependent responses and the dynamic nature of silencing events, shedding light on TRV mobility across different plant tissues.
病毒诱导的基因沉默（VIGS）是植物科学中一种多功能工具，但由于转化挑战，将其应用于向日葵等非模式物种需要进行大量优化。在本研究中，我们旨在阐明显著影响向日葵农杆菌介导的VIGS效率的因素。在测试了多种方法后，我们得出结论，种子真空处理技术随后进行6小时共培养产生了最有效的VIGS结果。基因型依赖性分析揭示了不同向日葵基因型的感染率（62%-91%）和沉默症状传播情况各不相同。此外，我们还探究了烟草脆裂病毒（TRV）在VIGS感染的向日葵植株不同组织和区域中的移动性以及表型沉默表现（光漂白）。我们发现TRV的存在不一定局限于有明显沉默事件的组织。最后，延时观察表明，与成熟组织相比，光漂白斑点在幼嫩组织中扩散更为活跃。本研究不仅为向日葵提供了一个可靠的VIGS方案，还为基因型依赖性反应以及沉默事件的动态性质提供了有价值的见解，揭示了TRV在不同植物组织中的移动性。
