{"author":[{"id":"243413","first_name":"Benjamin Willem","orcid":"0000-0002-7201-4335","last_name":"Moorlach","orcid_put_code_url":"https://api.orcid.org/v2.0/0000-0002-7201-4335/work/171497549","full_name":"Moorlach, Benjamin Willem"},{"first_name":"Désirée","last_name":"Jakobs-Schönwandt","full_name":"Jakobs-Schönwandt, Désirée"},{"full_name":"Poranen, Minna","last_name":"Poranen","first_name":"Minna"},{"full_name":"Ladera Carmona, Maria José","last_name":"Ladera Carmona","first_name":"Maria José"},{"full_name":"Abdeldayem, Mohamed","last_name":"Abdeldayem","first_name":"Mohamed"},{"last_name":"Kogel","full_name":"Kogel, Karl-Heinz","first_name":"Karl-Heinz"},{"full_name":"Sede, Ana","last_name":"Sede","first_name":"Ana"},{"last_name":"Heinlein","full_name":"Heinlein, Manfred","first_name":"Manfred"},{"full_name":"Patel, Anant","last_name":"Patel","first_name":"Anant","id":"201870"}],"language":[{"iso":"eng"}],"date_updated":"2024-11-13T06:27:04Z","conference":{"location":"Georg-August-Universität Göttingen","name":"63. Deutsche Pflanzenschutztagung","end_date":"2023-09-29","start_date":"2023-09-26"},"abstract":[{"lang":"eng","text":"Growing concerns over negative effects of pesticides on the environment and on human health are the cause of an increased demand for environmentally sustainable plant protection agents. A promising substitute for conventional synthetic chemical pesticides is double stranded RNA (dsRNA). In general, dsRNA is a nature-derived and environmentally safe molecule that occurs in all organisms and is rapidly degraded in soils. Its application triggers RNA-induced gene silencing, also known as RNA interference (RNAi), an ancient cellular mechanism of eucarytotic hosts to protect themselves against pathogens (Rank & Koch, 2021).\r\nIn the ERA-NET project BioProtect, we are developing spray-induced gene silencing (SIGS) approaches with dsRNA artificially produced in a bacterial host system (Niehl et al., 2018) and formulated with biologically degradable components to stabilize and protect the dsRNA and improve its uptake in planta upon delivery to crops with conventional spraying techniques.  Here, we report on the design of dsRNAs that target specific viral and fungal plant pathogens. Some of the major hurdles of dsRNA application in agriculture are the degradation on the leaf surface, and the reduced uptake of the negatively charged dsRNA via the cuticle, cell wall and cell membrane and other plant barriers (Bennett et al., 2020). \r\nTo solve these problems, we developed a cationic dsRNA-biopolymer carrier formulation based on the electrostatic interaction of cationic and anionic biopolymers, which stabilizes and protects the dsRNA from degradation on the plant surface, and which increases uptake into the plant. To increase the uptake, biopolymers with appropriate characteristics, e.g. molecular weight and charge, were chosen and formulated with the optimal parameters, such as charge ratio, polymer concentration, and energy input, to achieve nanoparticle formulations with a positive zeta potential, indicating charge masking of dsRNA. The hydrodynamic diameter of the resulting formulation determined by dynamic light scattering was 76.4 ± 4.9 nm with a PDI of 0.175 ± 0.006. Scanning electron microscopy analysis confirmed the diameter and revealed that the formulation consists of spherical nanoparticles. Furthermore, energy-dispersive X-ray spectroscopy measurements verified dsRNA integration into the nanoparticles by the presence of a phosphorous peak in formulations. Moreover, electrophoretic light scattering measurements indicated that the nanoparticles had a positive zeta-potential of +20 to +50 mV, which confirms that the negative charge of dsRNA was successfully masked in the formulation.  \r\nIn the experiments on fungi, RNAi activity was determined by a reduction of infection sites and necrotic leaf area, as well as gene silencing measured by RT-qPCR. In the experiments against plant viruses, the RNAi was analysed by the number of infection sites, and viral accumulation of targeted-viruses was determined by RT-qPCR. Moreover, these data were compared to nanocarrier formulations of unspecific RNA sequences and without dsRNA, which indicated that both nanocarrier and unspecific dsRNA sequences activate PTI reponses, thereby reinforcing plant protection through a second host defence pathway.\r\nIn future, we plan to investigate the RNAi efficacy of formulated dsRNA designed against sap-sucking insects. Finally, we will develop a carrier liquid based on biologically degradable surfactants to deliver the formulation to agricultural crops in the field.\r\n\r\nLiterature:\r\nBennett, M., J. Deikman, B. Hendrix, A. Iandolino, 2020: Barriers to Efficient Foliar Uptake of dsRNA and Molecular Barriers to dsRNA Activity in Plant Cells. Front Plant Sci 11, 816, DOI: 10.3389/fpls.2020.00816.\r\nNiehl, A., M. Soininen, M.M. Poranen, M. Heinlein, 2018: Synthetic biology approach for plant protection using dsRNA. Plant Biotechnol J, DOI: 10.1111/pbi.12904.\r\nRank, A.P., A. Koch, 2021: Lab-to-Field Transition of RNA Spray Applications - How Far Are We? Frontiers in Plant Science 12, DOI: ARTN 755203\r\n10.3389/fpls.2021.755203.\r\n\r\n"}],"type":"conference","year":"2023","related_material":{"link":[{"url":"https://www.openagrar.de/servlets/MCRFileNodeServlet/openagrar_derivate_00055333/JKA_475_204.pdf","relation":"other"}]},"status":"public","_id":"5107","citation":{"ieee":"B. W. Moorlach <i>et al.</i>, “Cationic Nanoparticle Formulations of dsRNA for Sprayable Plant Protection in a Changing Climate,” presented at the 63. Deutsche Pflanzenschutztagung, Georg-August-Universität Göttingen, 2023.","chicago":"Moorlach, Benjamin Willem, Désirée Jakobs-Schönwandt, Minna Poranen, Maria José Ladera Carmona, Mohamed Abdeldayem, Karl-Heinz Kogel, Ana Sede, Manfred Heinlein, and Anant Patel. “Cationic Nanoparticle Formulations of DsRNA for Sprayable Plant Protection in a Changing Climate,” 2023.","apa":"Moorlach, B. W., Jakobs-Schönwandt, D., Poranen, M., Ladera Carmona, M. J., Abdeldayem, M., Kogel, K.-H., … Patel, A. (2023). Cationic Nanoparticle Formulations of dsRNA for Sprayable Plant Protection in a Changing Climate. Presented at the 63. Deutsche Pflanzenschutztagung, Georg-August-Universität Göttingen.","short":"B.W. Moorlach, D. Jakobs-Schönwandt, M. Poranen, M.J. Ladera Carmona, M. Abdeldayem, K.-H. Kogel, A. Sede, M. Heinlein, A. Patel, in: 2023.","alphadin":"<span style=\"font-variant:small-caps;\"><span style=\"font-variant:small-caps;\">Moorlach, Benjamin Willem</span> ; <span style=\"font-variant:small-caps;\">Jakobs-Schönwandt, Désirée</span> ; <span style=\"font-variant:small-caps;\">Poranen, Minna</span> ; <span style=\"font-variant:small-caps;\">Ladera Carmona, Maria José</span> ; <span style=\"font-variant:small-caps;\">Abdeldayem, Mohamed</span> ; <span style=\"font-variant:small-caps;\">Kogel, Karl-Heinz</span> ; <span style=\"font-variant:small-caps;\">Sede, Ana</span> ; <span style=\"font-variant:small-caps;\">Heinlein, Manfred</span> ; u. a.</span>: Cationic Nanoparticle Formulations of dsRNA for Sprayable Plant Protection in a Changing Climate. In: , 2023","bibtex":"@inproceedings{Moorlach_Jakobs-Schönwandt_Poranen_Ladera Carmona_Abdeldayem_Kogel_Sede_Heinlein_Patel_2023, title={Cationic Nanoparticle Formulations of dsRNA for Sprayable Plant Protection in a Changing Climate}, author={Moorlach, Benjamin Willem and Jakobs-Schönwandt, Désirée and Poranen, Minna and Ladera Carmona, Maria José and Abdeldayem, Mohamed and Kogel, Karl-Heinz and Sede, Ana and Heinlein, Manfred and Patel, Anant}, year={2023} }","ama":"Moorlach BW, Jakobs-Schönwandt D, Poranen M, et al. Cationic Nanoparticle Formulations of dsRNA for Sprayable Plant Protection in a Changing Climate. In: ; 2023.","mla":"Moorlach, Benjamin Willem, et al. <i>Cationic Nanoparticle Formulations of DsRNA for Sprayable Plant Protection in a Changing Climate</i>. 2023."},"user_id":"220548","date_created":"2024-11-12T10:11:58Z","title":"Cationic Nanoparticle Formulations of dsRNA for Sprayable Plant Protection in a Changing Climate"}