|
|
|
|
|
Brown planthopper E78 regulates moulting and ovarian development by interacting with E93
|
| ZHENG Shi-wen1*, JIANG Xiao-juan1*, MAO Yi-wen1, LI Yan1, GAO Han1, LIN Xin-da1, 2# |
1 College of Life Sciences, China Jiliang University, Hangzhou 310018, P.R.China
2 College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, P.R.China
|
|
|
|
|
摘要
褐飞虱（Nilaparvata lugens）是亚洲许多水稻种植区的主要迁飞性害虫。E78是核激素受体超家族的成员。E78在卵巢发育和早期胚胎发生的母体调节中起重要作用。本研究克隆了褐飞虱E78基因（NlE78），预测的氨基酸序列显示其含有两个保守结构域：NR-LBD和DBD。 定量PCR（qRT-PCR）结果显示NlE78在5龄若虫和雌成虫的卵巢中表达较高。下调NlE78的表达后，蜕皮失败率（33.2%）显著增加，且卵巢发育延迟。然而，同时下调NlE78与NlE93的表达后，羽化率显着增加（78.79%），卵巢发育情况与NlE78下调时相似但其发育并未延迟。免疫共沉淀实验表明，NlE78 与 NlE93 有蛋白-蛋白相互作用，NlE93 是已知的蜕皮激素信号通路的关键下游转录因子。通过免疫荧光进行的细胞定位实验结果发现，NlE78和NlE93均在细胞核中表达。该研究表明，NlE78 可能通过与 NlE93 的蛋白-蛋白相互作用调节卵巢发育和蜕皮。本研究对于开发基于新靶点的新型农药和新防治方法具有重要意义。
Abstract
The brown planthopper (Nilaparvata lugens) is the main migratory pest in many rice growing areas in Asia. E78 is a member of the nuclear hormone receptor superfamily which plays an important role in egg development and maternal regulation of early embryogenesis. In this study, brown planthopper E78 (NlE78) was cloned, and the predicted amino acid sequence showed that it contains two conserved domains: NR-LBD and DBD. qRT-PCR showed that the expression of NlE78 is high in the fifth instar nymphs and the ovaries of females. After downregulation of NlE78, the rate of moulting failure (33.2%) increased significantly, and ovarian development was delayed. However, when NlE78 was downregulated together with NlE93, the emergence rate increased significantly (78.79%), and ovarian development was similar to that when NlE78 was downregulated but not delayed. A co-immunoprecipitation experiment showed that NlE78 interacts with NlE93, a crucial downstream transcription factor of the ecdysone signalling pathway. Cellular localization by immunofluorescence revealed that NlE78 and NlE93 are expressed in the nucleus. This study indicates that NlE78 regulates ovarian development and moulting, possibly through its interaction with NlE93. This study is of great significance for the development of new pesticides and control methods based on newly discovered targets.
|
|
Received: 28 April 2022
Accepted: 11 July 2022
|
| Fund:
This research was supported by the National Natural Science Foundation of China (32172390, 1672023 and 31741107) and the Natural Science Foundation of Zhejiang Province, China (LZ20C140002).
|
| About author: ZHENG Shi-wen, E-mail: 1186842121@qq.com; JIANG Xiao-juan, E-mail: 2293022580@qq.com; #Correspondence LIN Xin-da, Mobile: +86-13958028822, E-mail: linxinda@zjut.edu.cn
* These authors contributed equally to this study. |
Cite this article:
ZHENG Shi-wen, JIANG Xiao-juan, MAO Yi-wen, LI Yan, GAO Han, LIN Xin-da.
2023.
Brown planthopper E78 regulates moulting and ovarian development by interacting with E93
. Journal of Integrative Agriculture, 22(5): 1455-1464.
|
Ables E T, Bois K E, Garcia C A, Drummond-Barbosa D. 2015. Ecdysone response gene E78 controls ovarian germline stem cell niche formation and follicle survival in Drosophila. Developmental Biology, 400, 33–42.
Ables E T, Drummond-Barbosa D. 2010. The steroid hormone ecdysone functions with intrinsic chromatin remodeling factors to control female germline stem cells in Drosophila. Cell Stem Cell, 7, 581–592.
Ashburner M. 1974. Sequential gene activation by ecdysone in polytene chromosomes of Drosophila melanogaster: II. The effects of inhibitors of protein synthesis. Developmental Biology, 39, 141–157.
Bao Y Y, Li B L, Liu Z B, Xue J, Zhu Z R, Cheng J A, Zhang C X. 2010. Triazophos up-regulated gene expression in the female brown planthopper, Nilaparvata lugens. Journal of Insect Physiology, 56, 1087–1094.
Belles X, Piulachs M D. 2015. Ecdysone signalling and ovarian development in insects: From stem cells to ovarian follicle formation. Biochimica et Biophysica Acta, 1849, 181–186.
Belles X, Santos C G. 2014. The MEKRE93 (Methoprene tolerant-Krüppel homolog 1-E93) pathway in the regulation of insect metamorphosis, and the homology of the pupal stage. Insect Biochemistry and Molecular Biology, 52, 60–68.
Chao A T, Guild G M. 1986. Molecular analysis of the ecdysterone-inducible 2B5 “early” puff in Drosophila melanogaster. The EMBO Journal, 5, 143–150.
Cheng J A, Zhu Z R. 2006. Analysis on the key factors causing the outbreak of brown planthopper in Yangtze area, China in 2005. Plant Protection, 32, 104. (in Chinese)
Evans R M, Mangelsdorf D J. 2014. Nuclear receptors, RXR, and the big bang. Cell, 157, 255–266.
Finger D S, Whitehead K M, Phipps D N, Ables E T. 2021. Nuclear receptors linking physiology and germline stem cells in Drosophila. Vitamins and Hormones, 116, 327–362.
Huet F, Ruiz C, Richards G. 1993. Puffs and PCR: The in vivo
dynamics of early gene expression during ecdysone responses in Drosophila. Development, 118, 613–627.
Jin M N, Xue J, Yao Y, Lin X D. 2014. Molecular characterization and functional analysis of Krüppel-homolog 1 (Kr-h1) in the brown planthopper, Nilaparvata lugens (Stål). Journal of Integrative Agriculture, 13, 1972–1981.
King-Jones K, Thummel C S. 2005. Nuclear receptors - A perspective from Drosophila. Nature Reviews Genetics, 6, 311–323.
Kliewer S A, Umesono K, Mangelsdorf D J, Evans R M. 1992. Retinoid X receptor interacts with nuclear receptors in retinoic acid, thyroid hormone and vitamin D3 signalling. Nature, 355, 446–449.
Koelle M R, Talbot W S, Segraves W A, Bender M T, Cherbas P, Hogness D S. 1991. The Drosophila EcR gene encodes an ecdysone receptor, a new member of the steroid receptor superfamily. Cell, 67, 59–77.
Lam G, Nam, H J, Velentzas P D, Baehrecke E H, Thummel C S. 2022. Drosophila E93 promotes adult development and suppresses larval responses to ecdysone during metamorphosis. Developmental Biology, 481, 104–115.
Lin X, Xu Y, Yao Y, Wang B, Lavine M D, Lavine L C. 2016a. JNK signaling mediates wing form polymorphism in brown planthoppers (Nilaparvata lugens). Insect Biochemistry and Molecular Biology, 73, 55–61.
Lin X, Xu Y L, Jiang J R, Lavine M, Lavine L C. 2018. Host quality induces phenotypic plasticity in a wing polyphenic insect. Proceedings of the National Academy of Sciences of the United States of America, 115, 7563–7568.
Lin X, Yao Y, Wang B, Lavine L C, Emlen D J. 2016b. Ecological trade-offs between migration and reproduction are mediated by the nutrition-sensitive insulin-signaling pathway. International Journal of Biological Sciences, 12, 607–616.
Liu X, Dai F, Guo E, Li K, Ma L, Tian L, Cao Y, Zhang G, Palli S R, Li S. 2015. 20-Hydroxyecdysone (20E) primary response gene E93 modulates 20E signaling to promote Bombyx larval-pupal metamorphosis. The Journal of Biological Chemistry, 290, 27370–27383.
Mao Y, Li Y, Gao H, Lin X. 2019. The direct interaction between E93 and Kr-h1 mediated their antagonistic effect on ovary development of the brown planthopper. International Journal of Molecular Sciences, 20, 2431.
Mou X, Duncan D M, Baehrecke E H, Duncan I. 2012. Control of target gene specificity during metamorphosis by the steroid response gene E93. Proceedings of the National Academy of Sciences of the United States of America, 109, 2949–2954.
Praggastis S A, Lam G, Horner M A, Nam H J, Thummel C S. 2021. The Drosophila E78 nuclear receptor regulates dietary triglyceride uptake and systemic lipid levels. Developmental Dynamics, 250, 640–651.
Russell S R, Heimbeck G, Goddard C M, Carpenter A T, Ashburner M. 1996. The Drosophila Eip78C gene is not vital but has a role in regulating chromosome puffs. Genetics, 144, 159–170.
Schmittgen T D, Livak K J. 2008. Analyzing real-time PCR data by the 2–∆∆CT method. Nature Protocols, 3, 1101–1108.
Sekimoto T, Iwami M, Sakurai S. 2007. 20-Hydroxyecdysone regulation of two isoforms of the Ets transcription factor E74 gene in programmed cell death in the silkworm anterior silk gland. Insect Molecular Biology, 16, 581–590.
Stilwell G E, Nelson C A, Weller J, Cui H, Hiruma K, Truman J W, Riddiford L M. 2003. E74 exhibits stage-specific hormonal regulation in the epidermis of the tobacco hornworm, Manduca sexta. Developmental Biology, 258, 76–90.
Stone B L, Thummel C S. 1993. The Drosophila 78C early late puff contains E78, an ecdysone-inducible gene that encodes a novel member of the nuclear hormone receptor superfamily. Cell, 75, 307–320.
Urena E, Manjon C, Franch-Marro X, Martin D. 2014. Transcription factor E93 specifies adult metamorphosis in hemimetabolous and holometabolous insects. Proceedings of the National Academy of Sciences of the United States of America, 111, 7024–7029.
Woodard C T, Baehrecke E H, Thummel C S. 1994. A molecular mechanism for the stage specificity of the Drosophila prepupal genetic response to ecdysone. Cell, 79, 607–615.
Yamanaka N, Rewitz K F, O’Connor M B. 2013. Ecdysone control of developmental transitions: Lessons from Drosophila research. Annual Reviews of Entomology, 58, 497–516.
|
| No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
