Double stranded RNAs as a modern tool for controlling plant pathogensJendrašić, Diego (Avtor) Jakše, Jernej (Mentor) Jeseničnik, Taja (Komentor) RNA interferenceHIGSSIGSsiRNAdsRNAplant pathogensRNA interference (RNAi) is a conserved biological mechanism that enables gene silencing by degradation of mRNA, offering a highly specific tool for plant pest and pathogen control. This thesis explores the application of double stranded RNAs (dsRNAs) in two broadly defined RNAi-based plant protection strategies: Host-Induced Gene Silencing (HIGS) and Spray-Induced Gene Silencing (SIGS). HIGS involves the transgenic expression of dsRNA in plants and has shown high efficacy against plant viruses, but it faces regulation, high cost and negative public perception. SIGS, a non-GMO alternative, delivers synthetic dsRNAs directly to plant surfaces, with its success depending on the formulation, stability, and biology of the target organism. This thesis examines important aspects of RNAi deployment, including design, production, and protection/stabilization. Emphasis is placed on target gene selection, avoidance of off-target effects, and the use of bioinformatics tools for high specificity. Production methods include in vitro transcription, microbial expression systems and cell-free platforms. Nanocarrier-based delivery systems protect dsRNAs from numerous environmental and biological factors. Calantha, the first commercial RNAi biopesticide and numerous trials against viral and fungal pathogens, illustrate the current progress and future potential of RNAi technology. These findings demonstrate RNAi-based technologies are a sustainable, precise, and environmentally safe approach to crop protection, with continuing innovations expected to address the remaining challenges related to field performance and regulatory adoption.20252025-08-24 07:15:52Diplomsko delo/naloga171378sl