نبذة مختصرة : RNA interference (RNAi) is a powerful tool for gene silencing. It has recently been used to design promising plant protection strategies against pests such as viruses, insects, etc. This generally requires modifying the plant genome to achieve in planta synthesis of the double-stranded RNA (dsRNA), which guides the cellular RNA interference machinery to silence the genes of interest. However, given Russian legislation, the approach in which dsRNA is synthesized by the plant itself remains unavailable for crop protection. The use of exogenously produced dsRNA appears to be a promising alternative, allowing researchers to avoid genetic modification of plants, making it possible to implement potential results in agriculture. Furthermore, exogenous dsRNAs are superior to chemical pesticides (fungicides, insecticides, etc.), which are widely used to control various plant diseases. The dsRNA acts through sequence-specific nucleic acid interactions, making it extremely selective and unlikely to harm off-target organisms. Thus, it seems promising to utilize RNAi technology for agricultural plant protection. In this case, questions arise regarding how to produce the required amounts of pathogen-specific exogenous dsRNA, and which delivery method will be optimal for providing sufficient protection. This work aims to utilize exogenous dsRNA to silence the Nicotiana benthamiana phytoene desaturase gene. Phytoene desaturase is a convenient model gene in gene silencing experiments, as its knockdown results in a distinct phenotypic manifestation, namely, leaf bleaching. The dsRNA synthesis for this work was performed in vivo in Escherichia coli cells, and the chosen delivery method was root treatment through watering, both techniques being as simple and accessible as possible. It is surmised that the proposed approach could be adapted for broader use of RNAi technologies in agricultural crop protection. ; РНК-интерференция – мощный инструмент для генного сайленсинга, благодаря чему используется при разработке новых подходов с ...
Relation: https://vavilov.elpub.ru/jour/article/view/4916/2030; Brigneti G., Martín-Hernández A.M., Jin H., Chen J., Baulcombe D.C., Baker B., Jones J.D.G. Virus-induced gene silencing in Solanum species. Plant J. 2004;39(2):264-272. doi 10.1111/J.1365-313X.2004.02122.x; Burch-Smith T.M., Schiff M., Liu Y., Dinesh-Kumar S.P. Efficient virus-induced gene silencing in Arabidopsis. Plant Physiol. 2006; 142(1):21-27. doi 10.1104/pp.106.084624; Carthew R.W., Sontheimer E.J. Origins and mechanisms of miRNAs and siRNAs. Cell. 2009;136(4):642-655. doi 10.1016/j.cell.2009.01.035; Dubrovina A.S., Kiselev K.V. Exogenous RNAs for gene regulation and plant resistance. Int J Mol Sci. 2019;20(9):2282. doi 10.3390/ijms20092282; Gan D., Zhang J., Jiang H., Jiang T., Zhu S., Cheng B. Bacterially expressed dsRNA protects maize against SCMV infection. Plant Cell Rep. 2010;29(11):1261-1268. doi 10.1007/s00299-010-0911-z; Jiang L., Ding L., He B., Shen J., Xu Z., Yin M., Zhang X. Systemic gene silencing in plants triggered by fluorescent nanoparticledelivered double-stranded RNA. Nanoscale. 2014;6(17):9965-9969. doi 10.1039/c4nr03481c; Kamthan A., Chaudhuri A., Kamthan M., Datta A. Small RNAs in plants: recent development and application for crop improvement. Front Plant Sci. 2015;6(APR). doi 10.3389/FPLS.2015.00208/PDF; Koch A., Biedenkopf D., Furch A., Weber L., Rossbach O., Abdel latef E., Linicus L., Johannsmeier J., Jelonek L., Goesmann A., Cardoza V., McMillan J., Mentzel T., Kogel K.H. An RNAi-based control of Fusarium graminearum infections through spraying of long dsRNAs involves a plant passage and is controlled by the fun gal silencing machinery. PLoS Pathog. 2016;12(10):e1005901. doi 10.1371/journal.ppat.1005901; Laurila M.R.L., Makeyev E.V., Bamford D.H. Bacteriophage φ6 RNA dependent RNA polymerase. Molecular details of initiating nucleic acid synthesis without primer. J Biol Chem. 2002;277(19):17117 17124. doi 10.1074/jbc.M111220200; Li H., Guan R., Guo H., Miao X. New insights into an RNAi approach for plant defence against piercing-sucking and stem-borer insect pests. Plant Cell Environ. 2015;38(11):2277-2285. doi 10.1111/pce.12546; Liu Y., Schiff M., Marathe R., Dinesh-Kumar S.P. Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for N-mediated re sistance to tobacco mosaic virus. Plant J. 2002;30(4):415-429. doi 10.1046/J.1365-313x.2002.01297.x; Mitter N., Worrall E.A., Robinson K.E., Li P., Jain R.G., Taochy C., Fletcher S.J., Carroll B.J., Lu G.Q., Xu Z.P. Clay nanosheets for topical delivery of RNAi for sustained protection against plant vi ruses. Nat Plants. 2017;3:16207. doi 10.1038/nplants.2016.207; Niehl A., Soininen M., Poranen M.M., Heinlein M. Synthetic biology approach for plant protection using dsRNA. Plant Biotechnol J. 2018;16(9):1679-1687. doi 10.1111/pbi.12904; Nishimura A., Morita M., Nishimura Y., Sugino Y. A rapid and highly efficient method for preparation of competent Escherichia coli cells. Nucleic Acids Res. 1990;18(20):6169. doi 10.1093/nar/18.20.6169; Numata K., Ohtani M., Yoshizumi T., Demura T., Kodama Y. Local gene silencing in plants via synthetic dsRNA and carrier peptide. Plant Biotechnol J. 2014;12(8):1027-1034. doi 10.1111/pbi.12208; Ratcliff F., Martin-Hernandez A.M., Baulcombe D.C. Tobacco rattle virus as a vector for analysis of gene function by silencing. Plant J. 2001;25(2):237-245. doi 10.1046/J.0960-7412.2000.00942.x; Tenllado F., Martínez-García B., Vargas M., Díaz-Ruíz J.R. Crude ex tracts of bacterially expressed dsRNA can be used to protect plants against virus infections. BMC Biotechnol. 2003;3:3. doi 10.1186/1472-6750-3-3; Tiwari I.M., Jesuraj A., Kamboj R., Devanna B.N., Botella J.R., Shar ma T.R. Host Delivered RNAi, an efficient approach to increase rice resistance to sheath blight pathogen (Rhizoctonia solani). Sci Rep. 2017;7(1):1-14. doi 10.1038/S41598-017-07749; Wang J., Gu L., Knipple D.C. Evaluation of some potential target genes and methods for RNAi-mediated pest control of the corn earworm Helicoverpa zea. Pestic Biochem Physiol. 2018;149:67-72. doi 10.1016/j.pestbp.2018.05.012; https://vavilov.elpub.ru/jour/article/view/4916
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