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Journal of Integrative Agriculture  2023, Vol. 22 Issue (2): 471-480    DOI: 10.1016/j.jia.2022.12.009
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CopE and TLR6 RNAi-mediated tomato resistance to western flower thrips

Jelli VENKATESH1*, Sung Jin KIM2*, Muhammad Irfan SIDDIQUE3, Ju Hyeon KIM4, Si Hyeock LEE2, Byoung-Cheorl KANG1

1 Department of Agriculture, Forestry and Bioresources, Research Institute of Agriculture and Life Sciences, Plant Genomics Breeding Institute, College of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea

2 Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Republic of Korea

3 Vegetable Research Division, National Institute of Horticulture and Herbal Science, Rural Development Administration, Jeonju 55365, Republic of Korea

4 Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea

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The western flower thrips (WFT; Frankliniella occidentalis) is a mesophyll cell feeder that damages many crops.  Management of WFT is complex due to factors such as high fecundity, short reproduction time, ability to feed on a broad range of host plants, and broad pesticide resistance.  These challenges have driven research into developing alternative pest control approaches for WFT.  This study analyzed the feasibility of a biological control-based strategy to manage WFT using RNA interference (RNAi)-mediated silencing of WTF endogenous genes.  For the delivery of RNAi, we developed transgenic tomato lines expressing double-stranded RNA (dsRNA) of coatomer protein subunit epsilon (CopE) and Toll-like receptor 6 (TLR6) from WFT.  These genes are involved in critical biological processes of WFT, and their dsRNA can be lethal to these insects when ingested orally.  Adult WFT that fed on the transgenic dsRNA-expressing tomato flower stalk showed increased mortality compared with insects that fed on wild-type samples.  In addition, WFT that fed on TLR6 and CopE transgenic tomato RNAi lines showed reduced levels of endogenous CopE and TLR6 transcripts, suggesting that their mortality was likely due to RNAi-mediated silencing of these genes.  Thus, our findings demonstrate that transgenic tomato plants expressing dsRNA of TLR6 and CopE can be lethal to F. occidentalis, suggesting that these genes may be deployed to control insecticide-resistant WFT.

Keywords:  Coatomer protein subunit Epsilon (CopE)        Frankliniella occidentalis        insect resistance        RNA interference        Toll-like receptor 6 (TLR6)         tomato        transgenics  
Received: 17 March 2022   Accepted: 10 October 2022

This research was supported by the Basic Science Research Program through the National Research Foundation (NRF), Ministry of Education, Korea (2021R1I1A1A01041938), a grant from the New Breeding Technologies Development Program, Rural Development Administration, Korea (PJ0165432022).  Mr. Sung Jin Kim was supported in part by the BK21 Plus Program, Ministry of Education, Korea.

About author:  Jelli VENKATESH, E-mail:; Correspondence Byoung-Cheorl KANG, Tel: +82-2-8804563; Fax: +82-2-8732056, E-mail:, * These authors contributed equally to this study.

Cite this article: 

Jelli VENKATESH, Sung Jin KIM, Muhammad Irfan SIDDIQUE, Ju Hyeon KIM, Si Hyeock LEE, Byoung-Cheorl KANG. 2023. CopE and TLR6 RNAi-mediated tomato resistance to western flower thrips. Journal of Integrative Agriculture, 22(2): 471-480.

Badillo-Vargas I E, Rotenberg D, Schneweis B A, Whitfield A E. 2015. RNA interference tools for the western flower thrips, Frankliniella occidentalis. Journal of Insect Physiology, 76, 36–46.
Barzman M, Bàrberi P, Birch A N, Boonekamp P, Dachbrodt-Saaydeh S, Graf B, Hommel B, Jensen J E, Kiss J, Kudsk P, Lamichhane J R. 2015. Eight principles of integrated pest management. Agronomy for Sustainable Development, 35, 1199–1215.
Baum J A, Bogaert T, Clinton W, Heck G R, Feldmann P, Ilagan O, Johnson S, Plaetinck G, Munyikwa T, Pleau M, Vaughn T. 2007. Control of coleopteran insect pests through RNA interference. Nature Biotechnology, 25, 1322–1326.
Bingsohn L, Knorr E, Billion A, Narva K, Vilcinskas A. 2017. Knockdown of genes in the Toll pathway reveals new lethal RNA interference targets for insect pest control. Insect Molecular Biology, 26, 92–102.
Bischoff V, Vignal C, Duvic B, Boneca I G, Hoffmann J A, Royet J. 2006. Downregulation of the Drosophila immune response by peptidoglycan-recognition proteins SC1 and SC2. PLoS Pathogens, 2, e14.
Bonifacino J S, Lippincott-Schwartz J. 2003. Coat proteins: Shaping membrane transport. Nature Reviews Molecular Cell Biology, 4, 409–414.
Chen S, Chen N, Miao B, Peng J, Zhang X, Chen C, Zhang X, Chang L, Du Q, Huang Y, Tong D. 2021. Coatomer protein COPε, a novel NS1-interacting protein, promotes the replication of Porcine Parvovirus via attenuation of the production of type I interferon. Veterinary Microbiology, 261, 109188.
Denecke S, Swevers L, Douris V, Vontas J. 2018. How do oral insecticidal compounds cross the insect midgut epithelium? Insect Biochemistry and Molecular Biology, 103, 22–35.
Doyle J J, Doyle J L. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, 19, 11–15.
Espinosa P J, Bielza P, Contreras J, Lacasa A. 2002. Insecticide resistance in field populations of Frankliniella occidentalis (Pergande) in Murcia (south-east Spain). Pest Management Science, 58, 967–971.
Ferry N, Edwards M, Gatehouse J, Capell T, Christou P, Gatehouse A. 2006. Transgenic plants for insect pest control: A forward looking scientific perspective. Transgenic Research, 15, 13–19.
Fujita K, Shimomura K, Yamamoto K I, Yamashita T, Suzuki K. 2006. A chitinase structurally related to the glycoside hydrolase family 48 is indispensable for the hormonally induced diapause termination in a beetle. Biochemical and Biophysical Research Communications, 345, 502–507.
Gao Y, Lei Z, Reitz S R. 2012. Western flower thrips resistance to insecticides: Detection, mechanisms and management strategies. Pest Management Science, 68, 1111–1121.
Hakim R S, Baldwin K, Smagghe G. 2010. Regulation of midgut growth, development, and metamorphosis. Annual Review of Entomolog, 55, 593–608.
Han S H, Kim J H, Kim K, Lee S H. 2019. Selection of lethal genes for ingestion RNA interference against western flower thrips, Frankliniella occidentalis, via leaf disc-mediated dsRNA delivery. Pesticide Biochemistry and Physiology, 161, 47–53.
Hossain M, Shimizu S, Matsuki M, Imamura M, Sakurai S, Iwami M. 2008. Expression of 20-hydroxyecdysone-induced genes in the silkworm brain and their functional analysis in post-embryonic development. Insect Biochemistry and Molecular Biology, 38, 1001–1007.
Huvenne H, Smagghe G. 2010. Mechanisms of dsRNA uptake in insects and potential of RNAi for pest control: A review. Journal of Insect Physiology, 56, 227–235.
Imler J L, Zheng L. 2004. Biology of Toll receptors: Lessons from insects and mammals. Journal of Leukocyte Biology, 75, 18–26.
Kawai T, Akira S. 2007. TLR signaling. Seminars in Immunology, 19, 24–32.
Kirk W D, Terry L I. 2003. The spread of the western flower thrips Frankliniella occidentalis (Pergande). Agricultural and Forest Entomology, 5, 301–310.
De Kogel W J, Koschier E H, Broughton S, Castañé C, Davidson M M, Hamilton J G, Kirk W D, Nielsen M C, Riudavets J, Van Tol R W, Teulon D A. 2015. Semiochemicals for sustainable thrips management. Bodenkultur, 66, 17–25.
Kwon D H, Park J H, Ashok P A, Lee U, Lee S H. 2016. Screening of target genes for RNAi in Tetranychus urticae and RNAi toxicity enhancement by chimeric genes. Pesticide Biochemistry and Physiology, 130, 1–7.
Kwon D H, Park J H, Lee S H. 2013. Screening of lethal genes for feeding RNAi by leaf disc-mediated systematic delivery of dsRNA in Tetranychus urticae. Pesticide Biochemistry and Physiology, 105, 69–75.
Lepelley A, Martin-Niclos M J, Le Bihan M, Marsh J A, Uggenti C, Rice G I, Bondet V, Duffy D, Hertzog J, Rehwinkel J, Amselem S. 2020. Mutations in COPA lead to abnormal trafficking of STING to the Golgi and interferon signaling. Journal of Experimental Medicine, 217, e20200600.
Mao Y B, Cai W J, Wang J W, Hong G J, Tao X Y, Wang L J, Huang Y P, Chen X Y. 2007. Silencing a cotton bollworm P450 monooxygenase gene by plant-mediated RNAi impairs larval tolerance of gossypol. Nature Biotechnology, 25, 1307–1313.
Meng X, Xie Z, Zhang N, Ji C, Dong F, Qian K, Wang J. 2018. Molecular cloning and characterization of GABA receptor and GluCl subunits in the western flower thrips, Frankliniella occidentalis. Pesticide Biochemistry and Physiology, 150, 33–39.
Miki D, Shimamoto K. 2004. Simple RNAi vectors for stable and transient suppression of gene function in rice. Plant Cell Physiology, 45, 490–495.
Miller S C, Brown S J, Tomoyasu Y. 2008. Larval RNAi in Drosophila? Development Genes and Evolution, 218, 505–510.
Minakuchi C, Namiki T, Shinoda T. 2009. Krüppel homolog 1, an early juvenile hormone-response gene downstream of Methoprene-tolerant, mediates its anti-metamorphic action in the red flour beetle Tribolium castaneum. Developmental Biology, 325, 341–350.
Mouden S, Sarmiento K F, Klinkhamer P G, Leiss K A. 2017. Integrated pest management in western flower thrips: Past, present and future. Pest Management Science, 73, 813–822.
Murata M, Konno K, Wasano N, Mochizuki A, Mitsuhara I. 2021. Expression of a gene for an MLX56 defense protein derived from mulberry latex confers strong resistance against a broad range of insect pests on transgenic tomato lines. PLoS ONE, 16, e0239958.
Ohnishi A, Hull J J, Matsumoto S. 2006. Targeted disruption of genes in the Bombyx mori sex pheromone biosynthetic pathway. Proceedings of the National Academy of Sciences of the United States of America, 103, 4398–4403.
Outchkourov N S, De Kogel W J, Wiegers G L, Abrahamson M, Jongsma M A. 2004. Engineered multidomain cysteine protease inhibitors yield resistance against western flower thrips (Frankliniella occidentalis) in greenhouse trials. Plant Biotechnology Journal, 2, 449–458.
Parthasarathy R, Palli S R. 2009. Molecular analysis of juvenile hormone analog action in controlling the metamorphosis of the red flour beetle, Tribolium castaneum. Archives of Insect Biochemistry and Physiology, 70, 57–70.
Pfaffl M W, Lange I, Daxenberger A, Meyer H H. 2001. Influence of an estrogen treatment on the tissue specific expression pattern of estrogen receptors (ER): Quantification of ER-alpha and ER-beta mRNA with real-time RT-PCR. Journal of Pathology, Microbiology and Immunolog, 109, 345–355.
Quan G, Kanda T, Tamura T. 2002. Induction of the white egg 3 
mutant phenotype by injection of the double-stranded RNA of the silkworm white gene. Insect Molecular Biology, 11, 217–222.
Reitz S R, Gao Y, Kirk W D, Hoddle M S, Leiss K A, Funderburk J E. 2020. Invasion biology, ecology, and management of western flower thrips. Annual Review of Entomology, 65, 17–37.
Roignant J Y, Carré C, Mugat B, Szymczak D, Lepesant J A, Antoniewski C. 2003. Absence of transitive and systemic pathways allows cell-specific and isoform-specific RNAi in Drosophila. RNA, 9, 299–308.
Song Y, An O, Ren X, Chan T H M, Tay D J T, Tang S J, Han J, Hong H, Ng V H, Ke X, Shen H. 2021. RNA editing mediates the functional switch of COPA in a novel mechanism of hepatocarcinogenesis. Journal of Hepatology, 74, 135–147.
Tauszig S, Jouanguy E, Hoffmann J A, Imler J L. 2000. Toll-related receptors and the control of antimicrobial peptide expression in Drosophila. Proceedings of the National Academy of Sciences of the United States of America, 97, 10520–10525.
Thomas J C, Wasmann C C, Echt C, Dunn R L, Bohnert H J, McCoy T J. 1994. Introduction and expression of an insect proteinase inhibitor in alfalfa Medicago sativa L. Plant Cell Reports, 14, 31–36.
Tomoyasu Y, Denell R E. 2004. Larval RNAi in Tribolium (Coleoptera) for analyzing adult development. Development Genes and Evolution, 214, 575–578.
Volpi S, Tsui J, Mariani M, Pastorino C, Caorsi R, Sacco O, Ravelli A, Shum A K, Gattorno M, Picco P. 2018. Type I interferon pathway activation in COPA syndrome. Clinical Immunology, 187, 33–36.
Weber A N, Tauszig-Delamasure S, Hoffmann J A, Lelièvre E, Gascan H, Ray K P, Morse M A, Imler J L, Gay N J. 2003. Binding of the Drosophila cytokine Spätzle to Toll is direct and establishes signaling. Nature Immunology, 4, 794–800.
Whitten M, Dyson P. 2017. Gene silencing in non-model insects: Overcoming hurdles using symbiotic bacteria for trauma-free sustainable delivery of RNA interference: Sustained RNA interference in insects mediated by symbiotic bacteria: Applications as a genetic tool and as a biocide. Bioessays, 39, 1600247.
Whitten M M, Facey P D, Del Sol R, Fernández-Martínez L T, Evans M C, Mitchell J J, Bodger O G, Dyson P J. 2016. Symbiont-mediated RNA interference in insects. Proceedings of the Royal Society (B: Biological Sciences), 283, 20160042.
Whyard S, Erdelyan C N, Partridge A L, Singh A D, Beebe N W, Capina R. 2015. Silencing the buzz: A new approach to population suppression of mosquitoes by feeding larvae double-stranded RNAs. Parasites & Vectors, 8, 1–11.
Wimmer E A. 2005. Eco-friendly insect management. Nature Biotechnology, 23, 432–433.
Xiong Y, Zeng H, Zhang Y, Xu D, Qiu D. 2013. Silencing the HaHR3 gene by transgenic plant-mediated RNAi to disrupt Helicoverpa armigera development. International Journal of Biological Sciences, 9, 370.
Yang T, Stoopen G, Thoen M, Wiegers G, Jongsma M A. 2013. Chrysanthemum expressing a linalool synthase gene ‘smells good’, but ‘tastes bad’to western flower thrips. Plant Biotechnology Journal, 11, 875–882.
Younis A, Siddique M I, Kim C K, Lim K B. 2014. RNA interference (RNAi) induced gene silencing: A promising approach of Hi-Tech plant breeding. International Journal of Biological Sciences, 10, 1150.
Yudin L, Cho J, Mitchell W. 1986. Host range of western flower thrips, Frankliniella occidentalis (Thysanoptera: Thripidae), with special reference to Leucaena glauca. Environmental Entomology, 15, 1292–1295.
Zha W, Peng X, Chen R, Du B, Zhu L, He G. 2011. Knockdown of midgut genes by dsRNA-transgenic plant-mediated RNA interference in the hemipteran insect Nilaparvata lugens. PLoS ONE, 6, e20504.
Zhu F, Xu J, Palli R, Ferguson J, Palli S R. 2011. Ingested RNA interference for managing the populations of the Colorado potato beetle, Leptinotarsa decemlineata. Pest Management Science, 67, 175–182.
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