Double stranded RNAs as a modern tool for controlling plant pathogens

Jendrašić, Diego (author), Jakše, Jernej (mentor), Jeseničnik, Taja (comentor)

RNA 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.

• Language: English
• Keywords: RNA interference, HIGS, SIGS, siRNA, dsRNA, plant pathogens
• Work type: Bachelor thesis/paper
• Typology: 2.11 - Undergraduate Thesis
• Organization: BF - Biotechnical Faculty
• Year: 2025
• PID: 20.500.12556/RUL-171378
• COBISS.SI-ID: 246628611
• Publication date in RUL: 24.08.2025

Secondary language

• Language: Slovenian
• Title: Dvo-verižne RNA kot sodobna strategija za zatiranje rastlinskih patogenov

Abstract

Interferenca RNA (RNAi) je ohranjen biološki mehanizem, ki omogoča utišanje genov z razgradnjo mRNA in predstavlja zelo specifično orodje za nadzor rastlinskih škodljivcev in patogenov. Ta diplomska naloga raziskuje uporabo dvo-verižnih RNA (dsRNA) v dveh različnih strategijah zaščite rastlin na osnovi RNAi: utišanje genov inducirano s strani gostitelja (HIGS), in utišanje genov inducirano pršenjem (SIGS). HIGS vključuje transgeno izražanje dsRNA v rastlinah in je pokazal visoko učinkovitost proti rastlinskim virusom, vendar se sooča z regulativnimi omejitvami, visokimi stroški in negativnim javnim mnenjem. SIGS, ne-GSO alternativa, dovaja sintetične dsRNA neposredno na površine rastlin, pri čemer je uspeh odvisen od formulacije, stabilnosti in biologije ciljnega organizma. Ta naloga obravnava pomembne vidike uporabe RNAi, vključno z načrtovanjem, proizvodnjo ter zaščito/stabilizacijo. Poudarek je na izbiri ciljnih genov, preprečevanju ne tarčnih učinkov in uporabi bioinformatičnih orodij za visoko specifičnost. Metode proizvodnje vključujejo in vitro transkripcijo, mikrobne ekspresijske sisteme in cell-free platforme. Dostavni sistemi na osnovi nanonosilcev ščitijo dsRNA pred številnimi okoljskimi in biološkimi dejavniki. Calantha, prvi komercialni biopesticid na osnovi RNAi, ter številni preizkusi proti virusnim in glivičnim patogenom ponazarjajo trenutni napredek in prihodnji potencial tehnologije RNAi. Ti izsledki kažejo, da so tehnologije na osnovi RNAi trajnosten, natančen in okolju varen pristop k zaščiti pridelkov, pri čemer se pričakuje, da bodo stalne inovacije naslovile preostale izzive, povezane s učinkovitostjo in regulativnim sprejemanjem.

• Keywords: RNA interferenca, HIGS, SIGS, siRNA, dsRNA, rastlinski patogeni