RNA interference (RNAi): A revolutionary approach for sustainable insect pest management
RNA INTERFERENCE: A REVOLUTIONARY APPROACH FOR SUSTAINABLE INSECT PEST MANAGEMENT
18 / 2
Authors
-
SNEHA SHARMA
Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221 005, Uttar Pradesh
-
VISHAW DEEP SHARMA
Dr. Yashwant Singh Parmar University of Horticulture and Forestry, Nauni, Solan-173230, Himachal Pradesh
-
M CHETHANKUMAR
Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221 005, Uttar Pradesh
-
RITTIK SARKAR
Institute of Agricultural Sciences, Banaras Hindu University, Varanasi-221 005, Uttar Pradesh
-
ARCHANA ANOKHE
ICAR-Indian Institute of Horticultural Research, Bengaluru-560089, Karnataka
-
P DURAIMURUGAN
ICAR-Indian Institute of Oilseeds Research, Rajendranagar, Hyderabad-500 030, Telangana
-
MANOJ KUMAR YADAV
ICAR-Directorate of Weed Research, Jabalpur-482002, Madhya Pradesh
Keywords:
Dicer, dsRNA, Gene silencing, Molecular pest control, RNAi, RISC, Sustainable agricultureAbstract
RNA interference (RNAi) is a naturally occurring gene regulatory mechanism present in most eukaryotic organisms and has emerged as a promising strategy for agricultural insect pest management. The process involves sequence-specific gene silencing mediated by double-stranded RNA (dsRNA) molecules that are complementary to the target messenger RNA (mRNA). In the RNAi pathway, specialized cellular enzymes recognize and cleave dsRNA into small interfering RNAs (siRNAs), which subsequently bind to the corresponding target mRNA and inhibit its expression. This suppression of essential genes disrupts vital physiological and developmental processes in insects. In recent years, extensive research has been conducted by scientific institutions, universities, and biotechnology industries to better understand the molecular mechanisms of RNAi and explore its practical applications in insect pest control. One of the major advantages of RNAi technology is its high degree of target specificity, allowing selective suppression of pest species with minimal impact on non-target organisms and beneficial insects, unlike conventional broad-spectrum chemical insecticides. Moreover, numerous studies have demonstrated the effectiveness of RNAi in inducing mortality and developmental abnormalities across various insect orders. Considering its precision, environmental safety, and potential efficacy, RNAi is regarded as a highly innovative molecular tool for sustainable and eco-friendly insect pest management.
Downloads
References
Abbasi R, Heschuk D, Kim B and Whyard S 2020. A novel paperclip double-stranded RNA structure demonstrates clathrin-independent uptake in the mosquito Aedes aegypti. Insect Biochemistry and Molecular Biology, 127: 103492. DOI: https://doi.org/10.1016/j.ibmb.2020.103492
Akami M, Njintang NY, Gbaye OA, Andongma AA, Rashid MA and Niu CY 2019. Gut bacteria of the cowpea beetle mediate its resistance to dichlorvos and susceptibility to Lippia adoensis essential oil. Scientific Reports, 9(1): 6435. DOI: https://doi.org/10.1038/s41598-019-42843-1
Ameen A and Raza S 2017. Green revolution: A review. International Journal of Advanced Scientific Research, 3(12): 129-137. DOI: https://doi.org/10.7439/ijasr.v3i12.4410
Araújo MF, Castanheira EM and Sousa SF 2023. The buzz on insecticides: A review of uses, molecular structures, targets, adverse effects, and alternatives. Molecules, 28(8): 3641. DOI: https://doi.org/10.3390/molecules28083641
Arimatsu Y, Kotani E, Sugimura Y and Furusawa T 2007. Molecular characterization of a cDNA encoding extracellular dsRNase and its expression in the silkworm, Bombyx mori. Insect Biochemistry and Molecular Biology, 37(2): 176-183. DOI: https://doi.org/10.1016/j.ibmb.2006.11.004
Baulcombe D 2004. RNA silencing in plants. Nature, 431(7006): 356-363. DOI: https://doi.org/10.1038/nature02874
Baum JA, Bogaert T, Clinton W, Heck GR, Feldmann P and Ilagan O 2007. Control of coleopteran insect pests through RNA interference. Nature Biotechnology, 25(11): 1322-1326. DOI: https://doi.org/10.1038/nbt1359
Bensoussan N, Dixit S, Tabara M, Letwin D, Milojevic M and Antonacci M 2020. Environmental RNA interference in two-spotted spider mite, Tetranychus urticae, reveals dsRNA processing requirements for efficient RNAi response. Scientific Reports, 10(1): 19126. DOI: https://doi.org/10.1038/s41598-020-75682-6
Cagliari D, Dias NP, Galdeano DM, Dos Santos EÁ, Smagghe G and Zotti MJ 2019. Management of pest insects and plant diseases by non-transformative RNAi. Frontiers in Plant Science, 10: 1319. DOI: https://doi.org/10.3389/fpls.2019.01319
Castellanos NL, Smagghe G, Sharma R, Oliveira EE and Christiaens O 2019. Liposome encapsulation and EDTA formulation of dsRNA targeting essential genes increase oral RNAi-caused mortality in the Neotropical stink bug Euschistus heros. Pest Management Science, 75(2): 537-548. DOI: https://doi.org/10.1002/ps.5167
Chao Z, Ma Z, Zhang Y, Yan S and Shen J 2022. RNA interference cannot be operated in lepidopteran insect? A nanocarrier breaks bottlenecks at all developmental stages of Spodoptera frugiperda, Research Square, DOI:10.21203/rs212785 DOI: https://doi.org/10.21203/rs.3.rs-2127285/v1
Chen A, Halilovic L, Shay JH, Koch A, Mitter N and Jin H 2023. Improving RNA-based crop protection through nanotechnology and insights from cross-kingdom RNA trafficking. Current Opinion in Plant Biology, 76: 102441. DOI: https://doi.org/10.1016/j.pbi.2023.102441
Chen J, Peng Y, Zhang H, Wang K, Zhao C and Zhu G 2021. Off-target effects of RNAi correlate with the mismatch rate between dsRNA and non-target mRNA. RNA Biology, 18(11): 1747-1759. DOI: https://doi.org/10.1080/15476286.2020.1868680
Chethankumar M, Sharma S, Anokhe A, Darshan GP and Uday Kumar BV 2025. RNA interference in pest management: A transformative approach. AgriCos e-Newsletter, 6(1): 10-13.
Christiaens O, Dzhambazova T, Kostov K, Arpaia S, Joga MR and Urru I 2018. Literature review of baseline information on RNAi to support the environmental risk assessment of RNAi-based GM plants. EFSA Supporting Publications, 15(5): 1424E. DOI: https://doi.org/10.2903/sp.efsa.2018.EN-1424
Christiaens O, Tardajos MG, Martinez Reyna ZL, Dash M, Dubruel P and Smagghe G 2018. Increased RNAi efficacy in Spodoptera exigua via the formulation of dsRNA with guanylated polymers. Frontiers in Physiology, 9: 316. DOI: https://doi.org/10.3389/fphys.2018.00316
Chung SH, Jing X, Luo Y and Douglas AE 2018. Targeting symbiosis-related insect genes by RNAi in the pea aphid-Buchnera symbiosis. Insect Biochemistry and Molecular Biology, 95: 55-63. DOI: https://doi.org/10.1016/j.ibmb.2018.02.004
Dalakouras A, Koidou V and Papadopoulou K 2024. DsRNA-based pesticides: Considerations for efficiency and risk assessment. Chemosphere, 352: 141530. DOI: https://doi.org/10.1016/j.chemosphere.2024.141530
Das J, Kumar R, Shah V and Sharma AK 2023. Simple cost-effective larval injection method for dsRNA delivery to induce RNAi response in Helicoverpa armigera. Journal of Applied Entomology, 147(4): 289-298. DOI: https://doi.org/10.1111/jen.13100
Das PK, Panda G, Patra K, Jena N and Dash M 2022. The role of polyplexes in developing a green, sustainable approach in agriculture. RSC Advances, 12(53): 34463-34481. DOI: https://doi.org/10.1039/D2RA06541J
European Parliament 2001. Directive 2001/18/EC of the European Parliament and of the Council of 12 March 2001 on the deliberate release into the environment of genetically modified organisms.
Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE and Mello CC 1998. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 391(6669): 806-811. DOI: https://doi.org/10.1038/35888
Fischer R 2012. Research agreements with Dow Agrosciences. Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V., Munich, Germany. Available from: https://www.fraunhofer.de/en/press/research-news/2012/january/research-agreement-with-dow-agrosciences. HTML.
Ghosh SK, Hunter WB, Park AL and Gundersen-Rindal DE 2017. Double-strand RNA delivery system for plant-sap feeding insects. PLoS ONE, 12(2): e0171861. DOI: https://doi.org/10.1371/journal.pone.0171861
Guan RB, Li HC, Fan YJ, Hu SR, Christiaens O and Smagghe G 2018. A nuclease specific to lepidopteran insects suppresses RNAi. Journal of Biological Chemistry, 293(16): 6011-6021. DOI: https://doi.org/10.1074/jbc.RA117.001553
Head GP, Carroll MW, Evans SP, Rule DM, Willse AR and Clark TL 2017. Evaluation of SmartStax and SmartStax PRO maize against western corn rootworm and northern corn rootworm: Efficacy and resistance management. Pest Management Science, 73(9): 1883-1899. DOI: https://doi.org/10.1002/ps.4554
Holmsgaard PN, Dealtry S, Dunon V, Heuer H, Hansen LH and Springael D 2017. Response of the bacterial community in an on-farm biopurification system, to which diverse pesticides are introduced over an agricultural season. Environmental Pollution, 229: 854-862. DOI: https://doi.org/10.1016/j.envpol.2017.07.026
Hough J, Howard JD, Brown S, Portwood DE, Kilby PM and Dickman MJ 2022. Strategies for the production of dsRNA biocontrols as alternatives to chemical pesticides. Frontiers in Bioengineering and Biotechnology, 10: 980592. DOI: https://doi.org/10.3389/fbioe.2022.980592
Jain RG, Fletcher SJ, Manzie N, Robinson KE, Li P and Lu E 2022. Foliar application of clay-delivered RNA interference for whitefly control. Nature Plants, 8(5): 535-548. DOI: https://doi.org/10.1038/s41477-022-01152-8
Jin H, Abouzaid M, Lin Y, Hull JJ and Ma W 2021. Cloning and RNAi-mediated three lethal genes that can be potentially used for Chilo suppressalis management. Pesticide Biochemistry and Physiology, 174: 104828. DOI: https://doi.org/10.1016/j.pestbp.2021.104828
Joga MR, Zotti MJ, Smagghe G and Christiaens O 2016. RNAi efficiency, systemic properties, and novel delivery methods for pest insect control: What we know so far. Frontiers in Physiology, 7: 553. DOI: https://doi.org/10.3389/fphys.2016.00553
Katoch R and Thakur N 2012. Insect gut nucleases: A challenge for RNA interference-mediated insect control strategies. International Journal of Biochemistry and Biotechnology, 1(8): 198-203.
Katoch R, Sethi A, Thakur N and Murdock LL 2013. RNAi for insect control: Current perspective and future challenges. Applied Biochemistry and Biotechnology, 171(4): 847-873. DOI: https://doi.org/10.1007/s12010-013-0399-4
Khajuria C, Ivashuta S, Wiggins E, Flagel L, Moar W and Pleau M 2018. Development and characterization of the first dsRNA-resistant insect population from western corn rootworm, Diabrotica virgifera virgifera LeConte. PLoS ONE, 13(5): e0197059. DOI: https://doi.org/10.1371/journal.pone.0197059
Khan AM, Ashfaq M, Khan AA, Naseem MT and Mansoor S 2018. Evaluation of potential RNA-interference-target genes to control cotton mealybug, Phenacoccus solenopsis. Insect Science, 25(5): 778-786. DOI: https://doi.org/10.1111/1744-7917.12455
Koch A, Höfle L, Werner BT, Imani J, Schmidt A and Jelonek L 2019. SIGS vs HIGS: A study on the efficacy of two dsRNA delivery strategies to silence Fusarium FgCYP51 genes in infected host and non-host plants. Molecular Plant Pathology, 20(12): 1636-1644. DOI: https://doi.org/10.1111/mpp.12866
Kumar Gadde A, Reddy AJ, Chowdary DD, Kumar DS and Rao CS 2023. RNA interference (RNAi)-based biopesticides. In Current development in biosciences & agriculture. India: National Press Associates, New Delhi, India.
Laisney J, Rose VL, Watters K, Donohue KV and Unrine JM 2021. Delivery of short hairpin RNA in the neotropical brown stink bug, Euschistus heros, using a composite nanomaterial. Pesticide Biochemistry and Physiology, 177: 104906. DOI: https://doi.org/10.1016/j.pestbp.2021.104906
Li BJ, Wang KK, Yu Y, Wei JQ, Zhu J and Wang JL 2023. PxRdl2 dsRNA increased the insecticidal activities of GABAR-targeting compounds against Plutella xylostella. Pesticide Biochemistry and Physiology, 195: 105548. DOI: https://doi.org/10.1016/j.pestbp.2023.105548
Linyu W, Lianjun Z, Ning L, Xiwu G and Xiaoning L 2021. Effect of RNAi targeting CYP6CY3 on the growth, development and insecticide susceptibility of Aphis gossypii using nanocarrier-based transdermal dsRNA delivery system. Pesticide Biochemistry and Physiology, 177: 104878. DOI: https://doi.org/10.1016/j.pestbp.2021.104878
Lomate PR and Bonning BC 2016. Distinct properties of proteases and nucleases in the gut, salivary gland and saliva of southern green stink bug, Nezara viridula. Scientific Reports, 6(1): 27587. DOI: https://doi.org/10.1038/srep27587
Lu Y, Deng X, Zhu Q, Wu D, Zhong J and Wen L 2023. The dsRNA delivery, targeting and application in pest control. Agronomy, 13(3): 714. DOI: https://doi.org/10.3390/agronomy13030714
Lucena-Leandro VS, Abreu EF, Vidal LA, Torres CR, Junqueira CI and Dantas J 2022. Current scenario of exogenously induced RNAi for lepidopteran agricultural pest control: From dsRNA design to topical application. International Journal of Molecular Sciences, 23(24): 15836. DOI: https://doi.org/10.3390/ijms232415836
Ma Z, Zhang Y, Li M, Chao Z, Du X and Yan S 2023. A first greenhouse application of bacteria-expressed and nanocarrier-delivered RNA pesticide for Myzus persicae control. Journal of Pest Science, 96(1): 181-193. DOI: https://doi.org/10.1007/s10340-022-01485-5
Malik HJ, Raza A, Amin I, Scheffler JA, Scheffler BE and Brown JK 2016. RNAi-mediated mortality of the whitefly through transgenic expression of double-stranded RNA homologous to acetylcholinesterase and ecdysone receptor in tobacco plants. Scientific Reports, 6(1): 38469. DOI: https://doi.org/10.1038/srep38469
Mat Jalaluddin NS, Othman RY and Harikrishna JA 2019. Global trends in research and commercialization of exogenous and endogenous RNAi technologies for crops. Critical Reviews in Biotechnology, 39(1): 67-78. DOI: https://doi.org/10.1080/07388551.2018.1496064
Mehlhorn S, Hunnekuhl VS, Geibel S, Nauen R and Bucher G 2021. Establishing RNAi for basic research and pest control and identification of the most efficient target genes for pest control: A brief guide. Frontiers in Zoology, 18: 1-6. DOI: https://doi.org/10.1186/s12983-021-00444-7
Meister G and Tuschl T 2004. Mechanisms of gene silencing by double-stranded RNA. Nature, 431(7006): 343-349. DOI: https://doi.org/10.1038/nature02873
Murphy KA, Tabuloc CA, Cervantes KR and Chiu JC 2016. Ingestion of genetically modified yeast symbiont reduces fitness of an insect pest via RNA interference. Scientific Reports, 6(1): 22587. DOI: https://doi.org/10.1038/srep22587
Nitnavare RB, Bhattacharya J, Singh S, Kour A, Hawkesford MJ and Arora N 2021. Next generation dsRNA-based insect control: Success so far and challenges. Frontiers in Plant Science, 12: 673576. DOI: https://doi.org/10.3389/fpls.2021.673576
OECD 2023. Considerations for the human health risk assessment of externally applied dsRNA-based pesticides (ENV/CBC/ M ON O 2 0 2 3 ). Available from: https://www.oecd.org/ en/publications/considerations-for-thehuman-health-risk-assessment-of-externally-applied-dsrna-based-pesticides_54852048-en.html.
Pallis S, Alyokhin A, Manley B, Rodrigues T, Barnes E and Narva K 2023. Effects of low doses of a novel dsRNA-based biopesticide (Calantha) on the Colorado potato beetle. Journal of Economic Entomology, 116(2): 456-461. DOI: https://doi.org/10.1093/jee/toad034
Price DR and Gatehouse JA 2008. RNAi-mediated crop protection against insects. Trends in Biotechnology, 26(7): 393-400. DOI: https://doi.org/10.1016/j.tibtech.2008.04.004
Qiao H, Zhao J, Wang X, Xiao L, Zhu-Salzman K and Lei J 2023. An oral dsRNA delivery system based on chitosan induces G protein-coupled receptor kinase 2 gene silencing for Apolygus lucorum control. Pesticide Biochemistry and Physiology, 194: 105481. DOI: https://doi.org/10.1016/j.pestbp.2023.105481
Qiao Z, Ye Y, Chang PH, Thirunarayanan D and Wigginton KR 2018. Nucleic acid photolysis by UV254 and the impact of virus encapsidation. Environmental Science & Technology, 52(18): 10408-10415. DOI: https://doi.org/10.1021/acs.est.8b02308
Ramkumar G, Asokan R, Prasannakumar NR, Kariyanna B, Karthi S and Alwahibi MS 2021. RNA interference suppression of v-ATPase B and juvenile hormone binding protein genes through topically applied dsRNA on tomato leaves for control of Tuta absoluta. Frontiers in Physiology, 12: 742871. DOI: https://doi.org/10.3389/fphys.2021.742871
Romeis J and Widmer F 2020. Assessing the risks of topically applied dsRNA-based products to non-target arthropods. Frontiers in Plant Science, 11: 679. DOI: https://doi.org/10.3389/fpls.2020.00679
Rupawate PS, Roylawar P, Khandagale K, Gawande S, Ade AB and Jaiswal DK 2023. Role of gut symbionts of insect pests: A novel target for insect-pest control. Frontiers in Microbiology, 14: 1146390. DOI: https://doi.org/10.3389/fmicb.2023.1146390
Sachin GP 2019. Gene silencing-An effective tool in insect pest management. Available from:http://14.139.185.57:8080/jspui/bitstream/123456789/13745/1/sachin%20final%20report.pdf.
Siomi H and Siomi MC 2009. On the road to reading the RNA-interference code. Nature, 457(7228): 396-404. DOI: https://doi.org/10.1038/nature07754
Sogalad MS, Ushakiran B and Dinesh Kumar V. 2024. Enhancing efficiency and precision in CRISPR genome editing for plants using computational tools. Journal of Oilseeds Research, 41(3&4): 89-108. DOI: https://doi.org/10.56739/baqa6j43
Subramanian KS 2023. Nanotechnologies for vegetable oilseeds production. Journal of Oilseeds Research, 40(3): 98-104. DOI: https://doi.org/10.56739/qpawa413
Thairu MW, Skidmore IH, Bansal R, Nováková E, Hansen TE and Li-Byarlay H 2017. Efficacy of RNA interference knockdown using aerosolized short interfering RNAs bound to nanoparticles in three diverse aphid species. Insect Molecular Biology, 26(3): 356-368. DOI: https://doi.org/10.1111/imb.12301
Tomoyasu Y, Miller SC, Tomita S, Schoppmeier M, Grossmann D and Bucher G 2008. Exploring systemic RNA interference in insects: A genome-wide survey for RNAi genes in Tribolium. Genome Biology, 9(1): R10. DOI: https://doi.org/10.1186/gb-2008-9-1-r10
Upadhyay SK, Chandrashekar K, Thakur N, Verma PC, Borgio JF and Singh PK 2011. RNA interference for the control of whiteflies (Bemisia tabaci) by oral route. Journal of Biosciences, 36: 153-161. DOI: https://doi.org/10.1007/s12038-011-9009-1
Verhaeghe L 2021. Evaluating different double-stranded RNA structures for their ability to control pest flea beetles. Available from: http://hdl.handle.net/1993/37814.
Vimala Devi PS, Duraimurugan P, Chandrika PKSV, Vineela V and Hari PP. 2020. Novel formulations of Bacillus thuringiensis var. kurstaki: An eco-friendly approach for management of lepidopteran pests. World Journal of Microbiology and Biotechnology. DOI: https://doi.org/10.1007/s11274-020-02849-8
Whitfield R, Anastasaki A, Truong NP, Cook AB, Omedes-Pujol M and Loczenski Rose V 2018. Efficient binding, protection, and self-release of dsRNA in soil by linear and star cationic polymers. ACS Macro Letters, 7(8): 909-915. DOI: https://doi.org/10.1021/acsmacrolett.8b00420
Wu M, Dong Y, Zhang Q, Li S, Chang L and Loiacono FV 2022. Efficient control of western flower thrips by plastid-mediated RNA interference. Proceedings of the National Academy of Sciences, 119(15): e2120081119. DOI: https://doi.org/10.1073/pnas.2120081119
Xu L, Xu S, Sun L, Zhang Y, Luo J and Bock R 2021. Synergistic action of the gut microbiota in environmental RNA interference in a leaf beetle. Microbiome, 9: 1-14. DOI: https://doi.org/10.1186/s40168-021-01066-1
Yan S, Qian J, Cai C, Ma Z, Li J and Yin M 2020. Spray method application of transdermal dsRNA delivery system for efficient gene silencing and pest control on soybean aphid Aphis glycines. Journal of Pest Science, 93: 449-459. DOI: https://doi.org/10.1007/s10340-019-01157-x
Yang J, Zhang Y, Zhang Z, Ren M, Wang Y and Duan Y 2024. The development of an egg-soaking method for delivering dsRNAs into spider mites. Pesticide Biochemistry and Physiology, 201: 105905. DOI: https://doi.org/10.1016/j.pestbp.2024.105905
Yang S, Zou Z, Xin T, Cai S, Wang X and Zhang H 2022. Knockdown of hexokinase in Diaphorina citri by RNAi inhibits chitin synthesis and leads to abnormal phenotypes. Pest Management Science, 78(10): 4303-4313. DOI: https://doi.org/10.1002/ps.7049
Yang W, Wang B, Lei G, Chen G and Liu D 2022. Advances in nanocarriers to improve the stability of dsRNA in the environment. Frontiers in Bioengineering and Biotechnology, 10: 974646. DOI: https://doi.org/10.3389/fbioe.2022.974646
Yoon JS, Mogilicherla K, Gurusamy D, Chen X, Chereddy SC and Palli SR 2018. Double-stranded RNA binding protein, Staufen, is required for the initiation of RNAi in coleopteran insects. Proceedings of the National Academy of Sciences, 115(33): 8334-8339. DOI: https://doi.org/10.1073/pnas.1809381115
Zhang J, Khan SA, Hasse C, Ruf S, Heckel DG and Bock R 2015. Full crop protection from an insect pest by expression of long double-stranded RNAs in plastids. Science, 347(6225): 991-994. DOI: https://doi.org/10.1126/science.1261680
Zhang Y, Ke Z, Xu L, Yang Y, Chang L and Zhang J 2024. A faster killing effect of plastid-mediated RNA interference on a leaf beetle through induced dysbiosis of gut bacteria. Plant Communications, 5(9): 100974. DOI: https://doi.org/10.1016/j.xplc.2024.100974
