Al-Roshdi Maha R, Ammara Ume, Khan Jamal, Al-Sadi Abdullah M, Shahid Muhammad Shafiq
Department of Plant Sciences, College of Agricultural and Marine Sciences, Sultan Qaboos University, Al-Khod, Oman.
Front Plant Sci. 2023 Mar 30;14:1164921. doi: 10.3389/fpls.2023.1164921. eCollection 2023.
Tomato yellow leaf curl virus (TYLCV) is a global spreading begomovirus that is exerting a major restraint on global tomato production. In this transgenic approach, an RNA interference (RNAi)-based construct consisting of sequences of an artificial microRNA (amiRNA), a group of small RNA molecules necessary for plant cell development, signal transduction, and stimulus to biotic and abiotic disease was engineered targeting the AC1/Rep gene of the Oman strain of TYLCV-OM. The Rep-amiRNA constructs presented an effective approach in regulating the expression of the Rep gene against TYLCV as a silencing target to create transgenic L. plant tolerance against TYLCV infection. Molecular diagnosis by PCR followed by a Southern hybridization analysis were performed to confirm the effectiveness of agrobacterium-mediated transformation in T0/T1-transformed plants. A substantial decrease in virus replication was observed when T1 transgenic tomato plants were challenged with the TYLCV-OM infectious construct. Although natural resistance options against TYLCV infection are not accessible, the current study proposes that genetically transformed tomato plants expressing amiRNA could be a potential approach for engineering tolerance in plants against TYLCV infection and conceivably for the inhibition of viral diseases against different strains of whitefly-transmitted begomoviruses in Oman.
番茄黄化曲叶病毒（TYLCV）是一种在全球传播的双生病毒，对全球番茄生产造成了重大限制。在这种转基因方法中，构建了一种基于RNA干扰（RNAi）的结构，其由人工微小RNA（amiRNA）的序列组成，amiRNA是植物细胞发育、信号转导以及对生物和非生物病害刺激所必需的一组小RNA分子，该结构靶向TYLCV - OM阿曼株系的AC1/Rep基因。Rep - amiRNA结构为调控Rep基因表达提供了一种有效方法，以TYLCV作为沉默靶点，从而使转基因番茄对TYLCV感染产生耐受性。通过PCR进行分子诊断，随后进行Southern杂交分析，以确认农杆菌介导的转化在T0/T1转化植株中的有效性。当用TYLCV - OM感染性构建体对T1转基因番茄植株进行挑战时，观察到病毒复制显著减少。尽管目前尚无针对TYLCV感染的天然抗性选择，但当前研究表明，表达amiRNA的转基因番茄植株可能是一种在植物中培育对TYLCV感染耐受性的潜在方法，并且有可能用于抑制阿曼不同株系粉虱传播的双生病毒引起的病毒性病害。
