Hostname: page-component-848d4c4894-p2v8j Total loading time: 0.001 Render date: 2024-06-03T06:43:43.429Z Has data issue: false hasContentIssue false
  • English
  • Français

RNAi of vacuolar-type H+-ATPase genes causes growth delay and molting defect in Henosepilachna vigintioctopunctata

Published online by Cambridge University Press:  11 June 2021

Jie Zeng ,
Li-Li Mu ,
Lin Jin ,
Ahmad Ali Anjum  and
Guo-Qing Li Open the ORCID record for Guo-Qing Li [Opens in a new window]
Jie Zeng
Affiliation:
Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing210095, China
Li-Li Mu
Affiliation:
Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing210095, China
Lin Jin
Affiliation:
Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing210095, China
Ahmad Ali Anjum
Affiliation:
Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing210095, China
Guo-Qing Li*
Affiliation:
Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing210095, China
*
Author for correspondence: Guo-Qing Li, Email: ligq@njau.edu.cn
Get access Rights & Permissions [Opens in a new window]

Abstract

Henosepilachna vigintioctopunctata is one of the most serious insect pests to a large number of nightshades and cucurbits. RNA interference (RNAi) triggered by double-stranded RNA (dsRNA) offers a reduced risk approach to control the beetle. Identification of amenable target genes and determination of appropriate life stage for dsRNA treatment are two critical steps in order to improve RNAi efficiency. In the present paper, we identified three vATPase genes, namely HvvATPaseC, HvvATPaseE and HvvATPaseH. We found that the three transcripts were widely expressed in the eggs, first- to fourth-instar larvae, prepupae, pupae and adults. They were abundantly transcribed in the hindgut and Malpighian tubules, in contrast to the epidermis and fat body. Three days' ingestion of dsvATPaseC, dsvATPaseE and dsvATPaseH by the fourth-instar larvae significantly decreased corresponding transcript level by 90.1, 88.9 and 97.2%, greatly reduced larval fresh weight by 28.0, 29.9 and 28.0%, and caused 66.7, 100 and 78.7% larval lethality respectively. Comparably, 3 days' exposure of the third-instar larvae to dsvATPaseC significantly reduced HvvATPaseC mRNA level by 89.5%, decreased approximately 80% of the larval fresh weight, and killed 100% of the treated larvae. Therefore, the three vATPase genes, especially HvvATPaseE, are potential amenable target genes and young larvae are more susceptible to dsRNA. Our findings will enable the development of the dsRNA-based pesticide to control H. vigintioctopunctata.

Keywords

Henosepilachna vigintioctopunctatalarval growthmolting defectRNA interferencevacuolar-type H+-ATPase
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 111 , Issue 6 , December 2021 , pp. 705 - 714
Check for updates
Copyright
Copyright © The Author(s), 2021. Published by Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Allan, AK, Du, J, Davies, SA and Dow, JA (2005) Genome-wide survey of V-ATPase genes in Drosophila reveals a conserved renal phenotype for lethal alleles. Physiological Genomics 22, 128–138.CrossRefGoogle ScholarPubMed
Araujo, RN, Santos, A, Pinto, FS, Gontijo, NF, Lehane, MJ and Pereira, MH (2006) RNA interference of the salivary gland nitrophorin 2 in the triatomine bug Rhodnius prolixus (Hemiptera: Reduviidae) by dsRNA ingestion or injection. Insect Biochemistry and Molecular Biology 36, 683–693.CrossRefGoogle ScholarPubMed
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, 176–183.CrossRefGoogle ScholarPubMed
Baum, JA, Bogaert, T, Clinton, W, Heck, GR, Feldmann, P, Ilagan, O, Johnson, S, Plaetinck, G, Munyikwa, T, Pleau, M, Vaughn, T and Roberts, J (2007) Control of coleopteran insect pests through RNA interference. Nature Biotechnology 25, 1322–1326.CrossRefGoogle ScholarPubMed
Bettencourt, R, Terenius, O and Faye, I (2002) Hemolin gene silencing by dsRNA injected into Cecropia pupae is lethal to next generation embryos. Insect Molecular Biology 11, 267–271.CrossRefGoogle ScholarPubMed
Bustin, SA, Benes, V, Garson, JA, Hellemans, J, Huggett, J, Kubista, M, Mueller, R, Nolan, T, Pfaffl, MW, Shipley, GL, Vandesompele, J and Wittwer, CT (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clinical Chemistry 55, 611–622.CrossRefGoogle ScholarPubMed
Cappelle, K, de Oliveira, CF, Van Eynde, B, Christiaens, O and Smagghe, G (2016) The involvement of clathrin-mediated endocytosis and two Sid-1-like transmembrane proteins in double-stranded RNA uptake in the Colorado potato beetle midgut. Insect Molecular Biology 25, 315–323.CrossRefGoogle ScholarPubMed
Chen, X, Tian, H, Zou, L, Tang, B, Hu, J and Zhang, W (2008) Disruption of Spodoptera exigua larval development by silencing chitin synthase gene A with RNA interference. Bulletin of Entomological Research 98, 613–619.CrossRefGoogle ScholarPubMed
Christiaens, O, Swevers, L and Smagghe, G (2014) DsRNA degradation in the pea aphid (Acyrthosiphon pisum) associated with lack of response in RNAi feeding and injection assay. Peptides 53, 307–314.CrossRefGoogle ScholarPubMed
Christiaens, O, Whyard, S, Vélez, AM and Smagghe, G (2020) Double-stranded RNA technology to control insect pests: current status and challenges. Frontiers in Plant Science 11, 451.CrossRefGoogle ScholarPubMed
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.CrossRefGoogle ScholarPubMed
Cooper, AM, Silver, K, Zhang, J, Park, Y and Zhu, KY (2019) Molecular mechanisms influencing efficiency of RNA interference in insects. Pest Management Science 75, 18–28.CrossRefGoogle ScholarPubMed
Eleftherianos, I, Millichap, PJ, Ffrench-Constant, RH and Reynolds, SE (2006a) RNAi suppression of recognition protein mediated immune responses in the tobacco hornworm Manduca sexta causes increased susceptibility to the insect pathogen Photorhabdus. Developmental and Comparative Immunology 30, 1099–1107.CrossRefGoogle Scholar
Eleftherianos, I, Marokhazi, J, Millichap, PJ, Hodgkinson, AJ, Sriboonlert, A, Ffrench-Constant, RH and Reynolds, SE (2006b) Prior infection of Manduca sexta with non-pathogenic Escherichia coli elicits immunity to pathogenic Photorhabdus luminescens: roles of immune-related proteins shown by RNA interference. Insect Biochemistry and Molecular Biology 36, 517–525.CrossRefGoogle Scholar
Eleftherianos, I, Xu, M, Yadi, H, Ffrench-Constant, RH and Reynolds, SE (2009) Plasmatocyte-spreading peptide (PSP) plays a central role in insect cellular immune defenses against bacterial infection. Journal of Experimental Biology 212, 1840–1848.CrossRefGoogle Scholar
Fu, K-Y, Guo, W-C, Lü, F-G, Liu, X-P and Li, G-Q (2014) Response of the vacuolar ATPase subunit E to RNA interference and four chemical pesticides in Leptinotarsa decemlineata (Say). Pesticide Biochemistry and Physiology 114, 16–23.CrossRefGoogle Scholar
Guo, W-C, Fu, K-Y, Yang, S, Li, X-X and Li, G-Q (2015) Instar-dependent systemic RNA interference response in Leptinotarsa decemlineata larvae. Pesticide Biochemistry and Physiology 123, 64–73.CrossRefGoogle ScholarPubMed
Guo, W, Lü, J, Guo, M, Chen, S, Qiu, B, Sang, W, Yang, C, Zhang, Y and Pan, H (2019) De novo transcriptome analysis reveals abundant gonad-specific genes in the ovary and testis of Henosepilachna vigintioctopunctata. International Journal of Molecular Sciences 20, E4084.CrossRefGoogle ScholarPubMed
Kennedy, S, Wang, D and Ruvkun, G (2004) A conserved siRNA-degrading RNase negatively regulates RNA interference in C. elegans. Nature 427, 645–649.CrossRefGoogle ScholarPubMed
Kitagawa, N, Mazon, H, Heck, AJ and Wilkens, S (2008) Stoichiometry of the peripheral stalk subunits E and G of yeast V1-ATPase determined by mass spectrometry. Journal of Biological Chemistry 283, 3329–3337.CrossRefGoogle Scholar
Li, C and Xia, Y (2012) Vacuolar ATPase subunit H is essential for the survival and moulting of Locusta migratoria manilensis. Insect Molecular Biology 21, 405–413.CrossRefGoogle ScholarPubMed
Li, HR, Khajuria, C, Rangasamy, M, Gandra, P, Woosely, A, Hasler, J, Schulenberg, G, Worden, S, Mcewan, R, Evans, C, Siegfried, BD and Narva, KE (2015) Long dsRNA but not siRNA initiates RNAi in western corn rootworm larvae and adults. Journal of Applied Entomology 139, 432–445.CrossRefGoogle Scholar
Liu, S, Ding, Z, Zhang, C, Yang, B and Liu, Z (2010) Gene knockdown by intro-thoracic injections of double-stranded RNA in the brown planthopper, Nilaparvata lugens. Insect Biochemistry and Molecular Biology 40, 666–671.CrossRefGoogle ScholarPubMed
Liu, FZ, Yang, B, Zhang, AH, Ding, DR and Wang, GR (2019) Plant-mediated RNAi for controlling Apolygus lucorum. Frontiers in Plant Science 10, 64.CrossRefGoogle ScholarPubMed
Liu, S, Jaouannet, M, Dempsey, DA, Imani, J, Coustau, C and Kogel, KH (2020) RNA-based technologies for insect control in plant production. Biotechnology Advances 39, 107463.CrossRefGoogle ScholarPubMed
Lomazzo, E, Hussmann, GP, Wolfe, BB, Yasuda, RP, Perry, DC and Kellar, KJ (2011) Effects of chronic nicotine on heteromeric neuronal nicotinic receptors in rat primary cultured neurons. Journal of Neurochemistry 119, 153–164.CrossRefGoogle ScholarPubMed
Lü, J, Chen, S, Guo, M, Ye, C, Qiu, B, Wu, J, Yang, C and Pan, H (2018) Selection and validation of reference genes for RT-qPCR analysis of the ladybird beetle Henosepilachna vigintioctomaculata. Frontiers in Physiology 9, 1614.CrossRefGoogle ScholarPubMed
Luo, Y, Chen, Q, Luan, J, Chung, SH, Van Eck, J, Turgeon, R and Douglas, AE (2017) Towards an understanding of the molecular basis of effective RNAi against a global insect pest, the whitefly Bemisia tabaci. Insect Biochemistry and Molecular Biology 88, 21–29.CrossRefGoogle ScholarPubMed
McGuire, C, Stransky, L, Cotter, K and Forgac, M (2017) Regulation of V-ATPase activity. Frontiers in Bioscience 22, 609–622.Google ScholarPubMed
Mrinal, N and Nagaraju, J (2008) Intron loss is associated with gain of function in the evolution of the gloverin family of antibacterial genes in Bombyx mori. Journal of Biological Chemistry 283, 23376–23387.CrossRefGoogle ScholarPubMed
Muench, SP, Huss, M, Song, CF, Phillips, C, Wieczorek, H, Trinick, J and Harrison, MA (2009) Cryo-electron microscopy of the vacuolar ATPase motor reveals its mechanical and regulatory complexity. Journal of Molecular Biology 386, 989–999.CrossRefGoogle ScholarPubMed
Muench, SP, Scheres, SH, Huss, M, Phillips, C, Vitavska, O, Wieczorek, H, Trinick, J and Harrison, MA (2014) Subunit positioning and stator filament stiffness in regulation and power transmission in the V1 motor of the Manduca sexta V-ATPase. Journal of Molecular Biology 426, 286–300.CrossRefGoogle ScholarPubMed
Prentice, K, Christiaens, O, Pertry, I, Bailey, A, Niblett, C, Ghislain, M, Gheysen, G and Smagghe, G (2017) RNAi-based gene silencing through dsRNA injection or ingestion against the African sweet potato weevil Cylas puncticollis (Coleoptera: Brentidae). Pest Management Science 73, 44–52.CrossRefGoogle Scholar
Rebijith, KB, Asokan, R, Ranjitha, HH, Rajendra, PBS, Krishna, V and Kumar, NKK (2015) Diet-delivered dsRNAs for juvenile hormone-binding protein and vacuolar ATPase-H implied their potential in the management of the melon aphid (Hemiptera: Aphididae). Environmental Entomology 45, 268–275.CrossRefGoogle Scholar
Rinkevich, FD and Scott, JG (2013) Limitations of RNAi of α6 nicotinic acetylcholine receptor subunits for assessing the in vivo sensitivity to spinosad. Insect Science 20, 101–108.CrossRefGoogle ScholarPubMed
Scott, JG, Michel, K, Bartholomay, LC, Siegfried, BD, Hunter, WB, Smagghe, G, Zhu, KY and Douglas, AE (2013) Towards the elements of successful insect RNAi. Journal of Insect Physiology 59, 1212–1221.CrossRefGoogle ScholarPubMed
Shukla, JN, Kalsi, M, Sethi, A, Narva, KE, Fishilevich, E, Singh, S, Mogilicherla, K and Palli, SR (2016) Reduced stability and intracellular transport of dsRNA contribute to poor RNAi response in lepidopteran insects. RNA Biology 13, 656–669.CrossRefGoogle ScholarPubMed
Singh, IK, Singh, S, Mogilicherla, K, Shukla, JN and Palli, SR (2017) Comparative analysis of double-stranded RNA degradation and processing in insects. Scientific Reports 7, 17059.CrossRefGoogle ScholarPubMed
Song, HF, Zhang, JQ, Li, DQ, Cooper, AMW, Silver, K, Li, T, Liu, X, Ma, E, Zhu, KY and Zhang, J (2017) A double-stranded RNA degrading enzyme reduces the efficiency of oral RNA interference in migratory locust. Insect Biochemistry and Molecular Biology 86, 68–80.CrossRefGoogle ScholarPubMed
Song, Q, Meng, B, Xu, H and Mao, Z (2020) The emerging roles of vacuolar-type ATPase-dependent lysosomal acidification in neurodegenerative diseases. Translational Neurodegeneration 9, 17.CrossRefGoogle ScholarPubMed
Spit, J, Philips, A, Wynant, N, Santos, D, Plaetinck, G and Broeck, JV (2017) Knockdown of nuclease activity in the gut enhances RNAi efficiency in the Colorado potato beetle, Leptinotarsa decemlineata, but not in the desert locust, Schistocerca gregaria. Insect Biochemistry and Molecular Biology 81, 103–116.CrossRefGoogle Scholar
Ulrich, J, Dao, VA, Majumdar, U, Schmitt-Engel, C, Schwirz, J, Schultheis, D, Ströhlein, N, Troelenberg, N, Grossmann, D, Richter, T, Dönitz, J, Gerischer, L, Leboulle, G, Vilcinskas, A, Stanke, M and Bucher, G (2015) Large scale RNAi screen in Tribolium reveals novel target genes for pest control and the proteasome as prime target. BMC Genomics 16, 674.CrossRefGoogle ScholarPubMed
Vélez, AM, Jurzenski, J, Matz, N, Zhou, X, Wang, H, Ellis, M and Siegfried, BD (2016) Developing an in vivo toxicity assay for RNAi risk assessment in honey bees, Apis mellifera L. Chemosphere 144, 1083–1090.CrossRefGoogle Scholar
Vitavska, O, Wieczorek, H and Merzendorfer, H (2003) A novel role for subunit C in mediating binding of the H+-V-ATPase to the actin cytoskeleton. Journal of Biological Chemistry 278, 18499–18505.CrossRefGoogle Scholar
Vitavska, O, Merzendorfer, H and Wieczorek, H (2005) The V-ATPase subunit C binds to polymeric F-actin as well as to monomeric G-actin and induces cross-linking of actin filaments. Journal of Biological Chemistry 280, 1070–1076.CrossRefGoogle Scholar
Vogel, E, Santos, D, Mingels, L, Verdonckt, TW and Broeck, JV (2019) RNA interference in insects: protecting beneficials and controlling pests. Frontiers in Physiology 9, 1912.CrossRefGoogle ScholarPubMed
Wang, K, Peng, Y, Pu, J, Fu, W, Wang, J and Han, Z (2016) Variation in RNAi efficacy among insect species is attributable to dsRNA degradation in vivo. Insect Biochemistry and Molecular Biology 77, 1–9.CrossRefGoogle ScholarPubMed
Whyard, S, Singh, AD and Wong, S (2009) Ingested double-stranded RNAs can act as species-specific insecticides. Insect Biochemistry and Molecular Biology 39, 824–832.CrossRefGoogle ScholarPubMed
Wieczorek, H, Brown, D, Grinstein, S, Ehrenfeld, J and Harvey, WR (1999) Animal plasma membrane energization by proton-motive V-ATPases. BioEssays 21, 637–648.3.0.CO;2-W>CrossRefGoogle ScholarPubMed
Wieczorek, H, Beyenbach, KW, Huss, M and Vitavska, O (2009) Vacuolar-type proton pumps in insect epithelia. Journal of Experimental Biology 212, 1611–1619.CrossRefGoogle ScholarPubMed
Wu, J-J, Mu, L-L, Kang, W-N, Ze, L-J, Shen, C-H, Jin, L, Anjum, AA and Li, G-Q (2021) RNA interference targeting ecdysone receptor blocks the larval-pupal transition in Henosepilachna vigintioctopunctata. Insect Science 2021(28), 419–429. https://doi.org/10.1111/1744-7917.12777.CrossRefGoogle Scholar
Wynant, N, Santos, D, Verdonck, R, Spit, J, Van Wielendaele, P and Broeck, JV (2014) Identification, functional characterization and phylogenetic analysis of double stranded RNA degrading enzymes present in the gut of the desert locust, Schistocerca gregaria. Insect Biochemistry and Molecular Biology 46, 1–8.CrossRefGoogle ScholarPubMed
Xiao, D, Gao, X, Xu, J, Liang, X, Li, Q, Yao, J and Zhu, KY (2015) Clathrin-dependent endocytosis plays a predominant role in cellular uptake of double-stranded RNA in the red flour beetle. Insect Biochemistry and Molecular Biology 60, 68–77.CrossRefGoogle Scholar
Xu, Q-Y, Deng, P, Li, A, Zhang, Q, Mu, L-L, Fu, K-Y, Guo, W-C and Li, G-Q (2019) Functional characterization of ultraspiracle in Leptinotarsa decemlineata using RNA interference assay. Insect Molecular Biology 28, 676–688.CrossRefGoogle ScholarPubMed
Xu, P, Ze, L-J, Kang, W-N, Wu, J-J, Jin, L, Ali, AA and Li, G-Q (2020) Functional divergence of white genes in Henosepilachna vigintioctopunctata revealed by RNA interference. Insect Molecular Biology 29, 466–476.Google ScholarPubMed
Yoon, JS, Shukla, JN, Gong, ZJ, Mogilicherla, K and Palli, SR (2016) RNA interference in the Colorado potato beetle, Leptinotarsa decemlineata: identification of key contributors. Insect Biochemistry and Molecular Biology 78, 78–88.CrossRefGoogle ScholarPubMed
Zhang, QL, Wang, F, Guo, J, Deng, XY, Chen, JY and Lin, LB (2018) Characterization of ladybird Henosepilachna vigintioctopunctata transcriptomes across various life stages. Scientific Data 5, 180093.CrossRefGoogle ScholarPubMed
Zhu, KY and Palli, SR (2020) Mechanisms, applications, and challenges of insect RNA interference. Annual Review of Entomology 65, 293–311.CrossRefGoogle ScholarPubMed
Zhu, F, Xu, J, Palli, R, Ferguson, J and Palli, SR (2011) Ingested RNA interference for managing the populations of the Colorado potato beetle, Leptinotarsa decemlineata. Pest Management Science 67, 175–182.CrossRefGoogle ScholarPubMed
Zhu, L, Zhang, W, Li, G, Sun, QZ, Wang, JJ, Smagghe, G and Jiang, HB (2019) Molecular characterization of ecdysis triggering hormone and its receptor in citrus red mite (Panonychus citri). Comparative Biochemistry and Physiology A Molecular and Integrative Physiology 230, 100–105.CrossRefGoogle Scholar
Zotti, M, Dos Santos, EA, Cagliari, D, Christiaens, O, Taning, CNT and Smagghe, G (2018) RNA interference technology in crop protection against arthropod pests, pathogens and nematodes. Pest Management Science 74, 1239–1250.CrossRefGoogle ScholarPubMed
Supplementary material: File

Zeng et al. supplementary material

Zeng et al. supplementary material 1

Download Zeng et al. supplementary material(File)
File 11.3 KB
Supplementary material: File

Zeng et al. supplementary material

Zeng et al. supplementary material 2

Download Zeng et al. supplementary material(File)
File 824.8 KB