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Oral delivery of double-stranded RNA induces prolonged and systemic gene knockdown in Metaseiulus occidentalis only after feeding on Tetranychus urticae

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Abstract

Metaseiulus (=Typhlodromus or Galendromus) occidentalis is an important biological control agent. Functional genomic studies on this predator have been hampered by the lack of reverse genetic tools such as RNA interference (RNAi). In the current study, we evaluated possible RNAi responses in M. occidentalis females by feeding double-stranded RNA (dsRNA) of RpL11, RpS2, RpL8, or Pros26.4 genes in 20 % sucrose solution. Females needed to subsequently feed on two-spotted spider mites (Tetranychus urticae) to elicit a nearly complete loss of egg production. The corresponding gene knockdown was robust, long-term, and was observed in the very few eggs produced (systemic or parental RNAi). Interestingly, dsRNA-mediated gene knockdown could not be induced if these predators were provided only the sucrose diet after ingesting dsRNAs; T. urticae had to be provided to elicit the RNAi response. However, the spider mite diet was not needed for sustaining the dsRNA-mediated gene knockdown once it commenced. Oral delivery of dsRNA will be a valuable tool for efficient genome-wide functional screens in this important predatory mite.

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References

  • Arakane Y, Hogenkamp DG, Zhu YC, Kramer KJ, Specht CA, Beeman RW, Kanost MR, Muthukrishnan S (2004) Characterization of two chitin synthase genes of the red flour beetle, Tribolium castaneum, and alternate exon usage in one of the genes during development. Insect Bioch Mol Biol 34:291–304

    Article  CAS  Google Scholar 

  • Aung KM, Boldbaatar D, Umemiya-Shirafuji R, Liao M, Xuan X, Suzuki H, Galay RL, Tanaka T, Fujisaki K (2011) Scavenger receptor mediates systemic RNA interference in ticks. PLoS ONE 6:e28407

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Baum JA, Bogaert T, Clinton W, Heck GR, Feldmann P, Ilagan O, Johnson S, Plaetinck G, Munyikwa T, Pleau M, Vaughn T, Roberts J (2007) Control of coleopteran insect pests through RNA interference. Nature Biotech 25:1322–1326

    Article  CAS  Google Scholar 

  • Bernstein E, Caudy AA, Hammond SM, Hannon GJ (2001) Role for a bidentate ribonuclease in the initiation step of RNA interference. Nature 409:363–366

    Article  CAS  PubMed  Google Scholar 

  • Boutros M, Kiger AA, Armknecht S, Kerr K, Hild M, Koch B, Haas SA, Heidelberg Fly Array Consortium, Paro R, Perrimon N (2004) Genome-wide RNAi analysis of growth and viability in Drosophila cells. Science 303:832–835

    Article  CAS  PubMed  Google Scholar 

  • Bucher G, Scholten J, Klingler M (2002) Parental RNAi in Tribolium (Coleoptera). Curr Biol 12:R85–R86

    Article  CAS  PubMed  Google Scholar 

  • Burand JP, Hunter WB (2013) RNAi: future in insect management. J Invertebr Pathol 112(Suppl):S68–S74

    Article  CAS  PubMed  Google Scholar 

  • Campbell EM, Budge GE, Bowman AS (2010) Gene-knockdown in the honey bee mite Varroa destructor by a non-invasive approach: studies on a glutathione S-transferase. Parasit Vectors 3:73

    Article  PubMed Central  PubMed  Google Scholar 

  • Chen X, Tian H, Zou L, Tang B, Hu J, Zhang W (2008) Disruption of Spodoptera exigua larval development by silencing chitin synthase gene A with RNA silence. Bull Entomol Res 98:613–619

    Article  CAS  PubMed  Google Scholar 

  • Crosby MA, Goodman JL, Strelets VB, Zhang P, Gelbart WM, FlyBase Consortium (2007) FlyBase: genomes by the dozen. Nuc Acids Res 35:D486–D491

    Article  CAS  Google Scholar 

  • Dietzl G, Chen D, Schnorrer F, Su KC, Barinova Y, Fellner M, Gasser B, Kinsey K, Oppel S, Scheiblauer S, Couto A, Marra V, Keleman K, Dickson BJ (2007) A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila. Nature 448:151–156

    Article  CAS  PubMed  Google Scholar 

  • Dzitoyeva S, Dimitrijevic N, Manev H (2003) Identification of a novel Drosophila gene, beltless, using injectable embryonic and adult RNA interference (RNAi). BMC Genom 4:33

    Article  Google Scholar 

  • Field RP, Hoy MA (1985) Diapause behavior of genetically-improved strains of the spider mite predator Metaseiulus occidentalis (Acarina: Phytoseiidae). Entomol Exp Appl 38:113–120

    Article  Google Scholar 

  • Field RP, Hoy MA (1986) Evaluation of genetically-improved strains of Metaseiulus occidentalis (Nesbitt) (Acarina: Phytoseiidae) for integrated control of spider mites on roses in greenhouses. Hilgardia 54(2):1–31

    Google Scholar 

  • Fire A, Xu SQ, Montgomery MK, Kostas SA, Driver SE, Mello CC (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811

    Article  CAS  PubMed  Google Scholar 

  • Garbian Y, Maori E, Kalev H, Shafir S, Sela I (2012) Bidirectional Transfer of RNAi between honey bee and Varroa destructor: Varroa gene silencing reduces Varroa population. PLoS Pathog 8(12):e1003035

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hammond SM (2005) Dicing and slicing: the core machinery of the RNA interference pathway. FEBS Lett 579:5822–5829

    Article  CAS  PubMed  Google Scholar 

  • Hoy MA (1985) Recent advances in genetics and genetic improvement of the Phytoseiidae. Annu Rev Entomol 30:345–370

    Article  Google Scholar 

  • Hoy MA, Cave FE (1986) Screening for thelytoky in the parahaploid phytoseiid, Metaseiulus occidentalis (Nesbitt). Exp Appl Acarol 2:273–276

    Article  Google Scholar 

  • Hoy MA, Standow KA (1982) Inheritance of resistance to sulfur in the spider mite predator Metaseiulus occidentalis. Entomol Exp Appl 31:316–323

    Article  Google Scholar 

  • Hoy MA, Yu F, Meyer JM, Tarazona OA, Jeyaprakash A, Wu K (2013) Transcriptome sequencing and annotation of the predatory mite Metaseiulus occidentalis (Acari: Phytoseiidae): a cautionary tale about possible contamination by prey sequences. Exp Appl Acarol 59(3):283–296

    Article  CAS  PubMed  Google Scholar 

  • Huvenne H, Smagghe G (2010) Mechanisms of dsRNA uptake in insects and potential of RNAi for pest control: a review. J Insect Physiol 56:227–235

    Article  CAS  PubMed  Google Scholar 

  • Jeyaprakash A, Hoy MA (2000) Long PCR improves Wolbachia DNA amplification: wsp sequences found in 76% of sixty-three arthropod species. Insect Mol Biol 9(4):393–405

    Article  CAS  PubMed  Google Scholar 

  • Kamau LM, Wright HW, Nisbet AJ, Bowman AS (2013) Development of an RNA-interference procedure for gene knockdown in the poultry red mite, Dermanyssus gallinae: Studies on histamine releasing factor and Cathepsin-D. Afr J Biotech 12:1350–1356

    CAS  Google Scholar 

  • Khila A, Grbić M (2007) Gene silencing in the spider mite Tetranychus urticae: dsRNA and siRNA parental silencing of the Distal-less gene. Dev Genes Evol 217:241–251

    Article  CAS  PubMed  Google Scholar 

  • Kurscheid S, Lew-Tabor AE, Rodriguez VM, Bruyeres AG, Doogan VJ, Munderloh UG, Guerrero FD, Barrero RA, Bellgard MI (2009) Evidence of a tick RNAi pathway by comparative genomics and reverse genetics screen of targets with known loss-of-function phenotypes in Drosophila. BMC Mol Biol 10:26

    Article  PubMed Central  PubMed  Google Scholar 

  • Kwon DH, Park JH, Lee SH (2013) Screening of lethal genes for feeding RNAi by leaf disc-mediated systematic delivery of dsRNA in Tetranychus urticae. Pest Biochem Physiol 105(1):69–75

    Article  CAS  Google Scholar 

  • Li X, Zhang M, Zhang H (2011) RNA interference of four genes in adult Bactrocera dorsalis by feeding their dsRNAs. PLoS ONE 6(3):e17788

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Liu PZ, Kaufman TC (2004) Hunchback is required for suppression of abdominal identity, and for proper germband growth and segmentation in the intermediate germband insect Oncopeltus fasciatus. Development 131:1515–1527

    Article  CAS  PubMed  Google Scholar 

  • Liu Q, Rand TA, Kalidas S, Du F, Kim HE, Smith DP, Wang X (2003) R2D2, a bridge between the initiation and effector steps of the Drosophila RNAi pathway. Science 301:1921–1925

    Article  CAS  PubMed  Google Scholar 

  • Liu J, Carmell MA, Rivas FV, Marsden CG, Thomson JM, Song JJ, Hammond SM, Joshua-Tor L, Hannon GJ (2004) Argonaute2 is the catalytic engine of mammalian RNAi. Science 305:1437–1441

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Lum L, Yao S, Mozer B, Rovescalli A, Kessler DV, Nirenberg M, Beachy PA (2003) Identification of Hedgehog pathway components by RNAi in Drosophila cultured cells. Science 299:2039–2045

    Article  CAS  PubMed  Google Scholar 

  • Lynch JA, Desplan C (2006) A method for parental RNA interference in the wasp Nasonia vitripennis. Nat Protoc 1:486–494

    Article  CAS  PubMed  Google Scholar 

  • Marco Antonio DS, Guidugli-Lazzarini KR, Do Nascimento AM, Simões ZL, Hartfelder K (2008) RNAi-mediated silencing of vitellogenin gene function turns honeybee (Apis mellifera) workers into extremely precocious foragers. Biomed Life Sci 95:952–961

    Google Scholar 

  • Meister G, Tuschl T (2004) Mechanisms of gene silencing by double-stranded RNA. Nature 431:343–349

    Article  CAS  PubMed  Google Scholar 

  • Mitchell RD 3rd, Ross E, Osgood C, Sonenshine DE, Donohue KV, Khalil SM, Thompson DM, Michael RR (2007) Molecular characterization, tissue-specific expression and RNAi knockdown of the first vitellogenin receptor from a tick. Insect Biochem Mol Biol 37(4):375–388

    Article  CAS  PubMed  Google Scholar 

  • Mito T, Sarashina I, Zhang H, Iwahashi A, Okamoto H, Miyawaki K, Shinmyo Y, Ohuchi H, Noji S (2005) Non-canonical functions of hunchback in segment patterning of the intermediate germ cricket Gryllus bimaculatus. Development 132:2069–2079

    Article  CAS  PubMed  Google Scholar 

  • Ozawa R, Nishimura O, Yazawa S, Muroi A, Takabayashi J, Arimura G (2012) Temperature-dependent, behavioural, and transcriptional variability of a tritrophic interaction consisting of bean, herbivorous mite, and predator. Mol Ecol 21(22):5624–5635

    Article  PubMed  Google Scholar 

  • Paim RMM, Araujo RN, Lehane MJ, Gontijo NF, Pereira MH (2013) Long-term effects and parental RNAi in the blood feeder Rhodnius prolixus (Hemiptera; Reduviidae). Insect Biochem Mol Biol doi: 10.1016/j.ibmb.2013.08.008

  • Pasquinelli A, Hunter S, Bracht J (2005) MicroRNAs: a developing story. Curr Opin Genet Dev 15:200–205

    Article  CAS  PubMed  Google Scholar 

  • Pellino JL, Sontheimer EJ (2003) R2D2 leads the silencing trigger to mRNA’s death star. Cell 115:132–133

    Article  CAS  PubMed  Google Scholar 

  • Roush RT, Hoy MA (1981) Laboratory, glasshouse, and field studies of artificially selected carbaryl resistance in Metaseiulus occidentals. J Econ Entomol 74:142–147

    Google Scholar 

  • Schluns H, Crozier RH (2007) Relish regulates expression of antimicrobial peptide genes in the honeybee, Apis mellifera, shown by RNA interference. Insect Mol Biol 16:753–759

    Article  CAS  PubMed  Google Scholar 

  • Schwager EE, Pechmann M, Feitosa NM, McGregor AP, Damen WGM (2009) hunchback functions as a segmentation gene in the spider Achaearanea tepidariorum. Curr Biol 19:1333–1340

    Article  CAS  PubMed  Google Scholar 

  • Scott JG, Michel K, Bartholomay LC, Siegfried BD, Hunter WB, Smagghe G, Zhu KY, Douglas AE (2013) Towards the elements of successful insect RNAi. J Insect Physiol 59(12):1212–1221

    Article  CAS  PubMed  Google Scholar 

  • Shakesby AJ, Wallace IS, Isaacs HV, Pritchard J, Roberts DM, Douglas AE (2009) A water-specific aquaporin involved in aphid osmoregulation. Insect Biochem Mol Biol 39:1–10

    Article  CAS  PubMed  Google Scholar 

  • Sharma PP, Schwager EE, Giribet G, Jockusch EL, Extavour CG (2013) Distal-less and dachshund pattern both plesiomorphic and apomorphic structures in chelicerates: RNA interference in the harvestman Phalangium opilio (Opiliones). Evol Develop 15:228–242

    Article  CAS  Google Scholar 

  • Soares CAG, Lima CMR, Dolan MC, Piesman J, Beard CB, Zeidner NS (2005) Capillary feeding of specific dsRNA induces silencing of the isac gene in nymphal Ixodes scapularis ticks. Insect Mol Biol 14:443–452

    Article  CAS  PubMed  Google Scholar 

  • Song JJ, Smith SK, Hannon GJ, Joshua-Tor L (2004) Crystal structure of Argonaute and its implications for RISC slicer activity. Science 305:1434–1437

    Article  CAS  PubMed  Google Scholar 

  • Surakasi VP, Mohamed AA, Kim Y (2011) RNA interference of beta 1 integrin subunit impairs development and immune responses of the beet armyworm, Spodoptera exigua. J Insect Physiol 57:1537–1544

    Article  CAS  PubMed  Google Scholar 

  • Suzuki Y, Truman JW, Riddiford LM (2008) The role of Broad in the development of Tribolium castaneum: implications for the evolution of the holometabolous insect pupa. Development 135:569–577

    Article  CAS  PubMed  Google Scholar 

  • Tabara H, Yigit E, Siomi H, Mello CC (2002) The dsRNA binding protein RDE-4 interacts with RDE-1, DCR-1, and a DExH-box helicase to direct RNAi in C. elegans. Cell 109:861–871

    Article  CAS  PubMed  Google Scholar 

  • Tian H, Peng H, Yao Q, Chen H, Xie Q, Tang B, Zhang W (2009) Developmental control of a lepidopteran pest Spodoptera exigua by ingestion of bacterial expressing dsRNA of a non-midgut gene. PLoS ONE 4:e6225

    Article  PubMed Central  PubMed  Google Scholar 

  • Timmons L, Fire A (1998) Specific interference by ingested dsRNA. Nature 395:854

    Article  CAS  PubMed  Google Scholar 

  • Timmons L, Tabara H, Mello CC, Fire AZ (2003) Inducible systemic RNA silencing in Caenorhabditis elegans. Mol Biol Cell 14:2972–2983

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Turner CT, Davy MW, MacDiarmid RM, Plummer KM, Birch NP, Newcomb RD (2006) RNA interference in the light brown apple moth, Epiphyas postvittana (Walker) induced by double-stranded RNA feeding. Insect Mol Biol 15:383–391

    Article  CAS  PubMed  Google Scholar 

  • Ulvila J, Parikka M, Kleino A, Sormunen R, Ezekowitz RA, Kocks C, Rämet M (2006) Double-stranded RNA is internalized by scavenger receptor-mediated endocytosis in Drosophila S2 cells. J Biol Chem 281(20):14370–14375

    Article  CAS  PubMed  Google Scholar 

  • Voinnet O, Baulcombe DC (1997) Systemic signaling in gene silencing. Nature 389:533

    Article  Google Scholar 

  • Wu K, Hoy MA (2012) Extended starvation reduced and eliminated Wolbachia, but not Cardinium, from Metaseiulus occidentalis females (Acari: Phytoseiidae): a need to reassess Wolbachia’s status in this predatory mite? J Invertebr Path 109:20–26

    Article  Google Scholar 

  • Yu Q, Lin T, Feng G, Yang K, Pang Y (2008) Functional analysis of the putative antiapoptotic genes, p49 and iap4, of Spodoptera litura nucleopolyhedrovirus with RNAi. J Gen Virol 89:1873–1880

    Article  CAS  PubMed  Google Scholar 

  • Yu N, Christiaens O, Liu J, Niu J, Cappelle K, Caccia S, Huvenne H, Smagghe G (2013) Delivery of dsRNA for RNAi in insects: an overview and future directions. Insect Sci 20:4–14

    Article  PubMed  Google Scholar 

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Acknowledgments

The research was supported in part by the Institute of Food and Agricultural Sciences of the University of Florida, and the Davies, Fischer and Eckes Endowment in Biological Control to M.A.H. We thank Godfrey Maina for assistance in rearing the colonies.

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  1. Department of Entomology and Nematology, University of Florida, PO Box 11620, Gainesville, FL, 32611, USA

    Ke Wu & Marjorie A. Hoy

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Correspondence to Ke Wu.

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Wu, K., Hoy, M.A. Oral delivery of double-stranded RNA induces prolonged and systemic gene knockdown in Metaseiulus occidentalis only after feeding on Tetranychus urticae . Exp Appl Acarol 63, 171–187 (2014). https://doi.org/10.1007/s10493-014-9772-4

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