RNAi technique was used to regulate Fusarium crown and root rot in tomatoes caused by F. oxysporum f. sp. radicis-lycopersici (FORL), by targeting three essential genes namely FoCYP51, FoChs1, and FoEF2, considering the fungicide site of action. The designed naked dsRNA was delivered on a hexagonal mono-dispersed and biodegradable layered double hydroxide (LDH) nanosheets with 30–90 nm diameter, providing high protection and stability for dsRNA with long term sustained release manner inside plant cells. The laboratory findings revealed the functional role of the three selected genes in reducing FORL's invasive growth on tomato fruits, indicating their involvement in FORL pathogenesis on tomatoes, and could be good targets for disease control using RNAi. Three different practical methods were applied for topical delivery of dsRNA into the plant cells: i) leaves spray; ii) petiole adsorption; and iii) root dipping, demonstrating three significant levels of spray-induced gene silencing (SIGS) efficacy in controlling FORL virulence infecting tomato plants. Significantly, topical spray of dsRNA delivered on LDH nanosheets provided Fusarium crown and root rot protection for at least 60 days. Obtained findings introduce critical information data on the potential to exploit spray-induced gene silencing (SIGS) as an effective alternative sustainable strategy for plant disease management. Given the ease design and production of biodegradable nano-delivery systems with high specificity and sustainability, and ease of applicability against many disease caused fungi. The use of target-specific dsRNA as an anti-fungal agent has a lot of promise and provide exceptional potential as a unique disease control and plant protection strategy.

使用RNA干扰技术来调节枯萎冠和根腐病造成番茄尖孢镰孢F。sp. 根茎番茄(FORL)，通过针对三个基本基因，即 FoCYP51、FoChs1 和 FoEF2，考虑到杀菌剂的作用位点。设计的裸dsRNA被递送到直径为30-90 nm的六角形单分散和可生物降解的层状双氢氧化物（LDH）纳米片上，以在植物细胞内的长期缓释方式为dsRNA提供高保护和稳定性。实验室研究结果揭示了三个选定基因在减少 FORL 对番茄果实的侵袭性生长方面的功能作用，表明它们参与了番茄的 FORL 发病机制，并且可能是使用 RNAi 控制疾病的良好目标。应用三种不同的实用方法将 dsRNA 局部递送到植物细胞中：i) 叶喷雾；ii) 叶柄吸附；iii) 浸根，证明了三种显着水平的喷雾诱导基因沉默（SIGS）在控制感染番茄植物的 FORL 毒力方面的功效。值得注意的是，在 LDH 纳米片上局部喷洒 dsRNA 可为镰刀菌提供至少 60 天的冠腐和根腐保护。获得的研究结果介绍了关于利用喷雾诱导基因沉默 (SIGS) 作为植物病害管理的有效替代可持续策略的潜力的关键信息数据。鉴于具有高度特异性和可持续性的可生物降解纳米递送系统的易于设计和生产，并且易于适用于许多由真菌引起的疾病。使用目标特异性 dsRNA 作为抗真菌剂有很大的希望，并提供作为独特的疾病控制和植物保护策略的非凡潜力。