Transgenic Research

, Volume 23, Issue 1, pp 145–152 | Cite as

Plant-mediated RNAi of a gap gene-enhanced tobacco tolerance against the Myzus persicae

Original Paper

Abstract

Plant-mediated RNAi has been developed as a powerful weapon in the fight against agricultural insect pests. The gap gene hunchback (hb) is of crucial importance in insect axial patterning and knockdown of hb is deforming and lethal to the next generation. The peach potato aphid, Myzus persicae (Sulzer), has many host plants and can be found throughout the world. To investigate the effect of plant-mediated RNAi on control of this insect, the hb gene in M. persicae was cloned, plant RNAi vector was constructed, and transgenic tobacco expressing Mphb dsRNA was developed. Transgenic tobacco had a different integration pattern of the transgene. Bioassays were performed by applying neonate aphids to homozygous transgenic plants in the T2 generation. Results revealed that continuous feeding of transgenic diet reduced Mphb mRNA level in the fed aphids and inhibited insect reproduction, indicating successful knockdown of the target gene in M. persicae by plant-mediated RNAi.

Keywords

RNAi hunchback Myzus persicae Transgenic tobacco  

References

  1. Annis B, Tamaki G, Berry RE (1981) Seasonal occurrence of wild secondary hosts of the green peach aphid, Myzus persicae (Sulzer), in agricultural systems in the Yakima Valley. Environ Entomol 10:307–312Google Scholar
  2. Baum J, Bogaert T, Clinton W, Heck G, Feldmann P (2007) Control of coleopteran insect pests through RNA interference. Nat Biotechnol 25:1322–1326PubMedCrossRefGoogle Scholar
  3. Chen J, Zhang D, Yao Q, Zhang J, Dong X, Tian H, Chen J, Zhang W (2010) Feeding-based RNA interference of a trehalose phosphate synthase gene in the brown planthopper, Nilaparvata lugens. Insect Mol Biol 19:777–786PubMedCrossRefGoogle Scholar
  4. Chougule NP, Bonning BC (2012) Toxins for transgenic resistance to Hemipteran pests. Toxins 4:405–429PubMedCentralPubMedCrossRefGoogle Scholar
  5. Dykxhoorn DM, Novina CD, Sharp PA (2003) Killing the messenger: short RNAs that silence gene expression. Nat Rev Mol Cell Biol 4:457–467PubMedCrossRefGoogle Scholar
  6. Dzitoyeva S, Dimitrijevic N, Manev H (2001) Intra-abdominal injection of double-stranded RNA into anesthetized adult Drosophila triggers RNA interference in the central nervous system. Mol Psychiatry 6:665–670PubMedCrossRefGoogle Scholar
  7. Fairbairn DJ, Cavallaro AS, Bernard M, Mahalinga-Iyer J, Graham MW, Botella JR (2007) Host-delivered RNAi: an effective strategy to silence genes in plant parasitic nematodes. Planta 226:1525–1533PubMedCrossRefGoogle Scholar
  8. Finkelstein R, Perrimon N (1990) The orthodenticle gene is regulated by bicoid and torso and specifies Drosophila head development. Nature 346:485–488PubMedCrossRefGoogle Scholar
  9. Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811PubMedCrossRefGoogle Scholar
  10. Gordon KH, Waterhouse PM (2007) RNAi for insect-proof plants. Nat Biotech 25:1231–1232CrossRefGoogle Scholar
  11. Hannon GJ (2002) RNA interference. Nature 418:244–251PubMedCrossRefGoogle Scholar
  12. Horsch RB, Fry JE, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231 CrossRefGoogle Scholar
  13. Huang T, Cook CE, Davis GK, Shigenobu S, Chen RPY, Chang C (2010) Anterior development in the parthenogenetic and viviparous form of the pea aphid, Acyrthosiphon pisum: hunchback and orthodenticle expression. Insect Mol Biol 19:75–85Google Scholar
  14. Huvenne H, Smagghe G (2010) Mechanisms of dsRNA uptake in insects and potential of RNAi for pest control: a review. J Insect Physiol 56:227–235PubMedCrossRefGoogle Scholar
  15. Jürgens G, Wieschaus E, Nüsslein-Volhard C, Kluding H (1984) Mutations affecting the pattern of the larval cuticle in Drosophila melanogaster. Roux’s Arch Dev Biol 193:283–295Google Scholar
  16. Lehmann R, Nüsslein-Volhard C (1987) hunchback, a gene required for segmentation of an anterior and posterior region of the Drosophila embryo. Dev Biol 119:402–417PubMedCrossRefGoogle Scholar
  17. Liu P, 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 131:1515–1527PubMedCrossRefGoogle Scholar
  18. Liu S, Ding Z, Zhang C, Yang B, Liu Z (2010) Gene knockdown by introthoracic injection of double-stranded RNA in the brown planthopper, Nilaparvata lugens. Insect Biochem Mol Biol 40:666–671PubMedCrossRefGoogle Scholar
  19. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  20. Lynch JA, Desplan C (2006) A method for parental RNA interference in the wasp Nasonia vitripennis. Nat Protoc 1:486–494PubMedCrossRefGoogle Scholar
  21. Lynch JA, Brent AE, Leaf DS, Pultz MA, Desplan C (2006) Localized maternal orthodenticle patterns anterior and posterior in the long germ wasp Nasonia. Nature 439:728–732PubMedCrossRefGoogle Scholar
  22. Mao J, Zeng F (2012) Feeding-based RNA intereference of a gap gene is lethal to the pea aphid, Acyrthosiphon pisum. PLoS ONE 7(11):e48718PubMedCentralPubMedCrossRefGoogle Scholar
  23. Mao Y, Cai W, Wang J, Hong G, Tao X, Wang L, Huang YP, Chen XY (2007) Silencing a cotton bollworm P450 monooxygenase gene by plant-mediated RNAi impairs larval tolerance of gossypol. Nat Biotech 25:1307–1313CrossRefGoogle Scholar
  24. Meister G, Tuschl T (2004) Mechanisms of gene silencing by double-stranded RNA. Nature 431:343–349PubMedCrossRefGoogle Scholar
  25. Meyering-Vos M, Muller A (2007) RNA interference suggests sulfakinins as satiety effectors in the cricket Gryllus bimaculatus. J Insect Physiol 53:840–848PubMedCrossRefGoogle Scholar
  26. Namba R, Sylvester ES (1981) Transmission of cauliflower mosaic virus by the green peach, turnip, cabbage, and pea aphids. J Econ Entomol 74:546–551Google Scholar
  27. Novotny T, Eiselt R, Urban J (2002) Hunchback is required for the specification of the early sublineage of neuroblast 7–3 in the Drosophila central nervous system. Development 129:1027–1036PubMedGoogle Scholar
  28. Patel NH, Hayward DC, Lall S, Pirkl NR, DiPietro D, Ball EE (2001) Grasshopper hunchback expression reveals conserved and novel aspects of axis formation and segmentation. Development 128:3459–3472PubMedGoogle Scholar
  29. Pitino M, Coleman AD, Maffei ME, Ridout CJ, Hogenhout SA (2011) Silencing of aphid genes by dsRNA feeding from plants. PLoS ONE 6:1–8CrossRefGoogle Scholar
  30. Price DRG, Gatehouse JA (2008) RNAi-mediated crop protection against insects. Trends Biotechnol 26:393–400PubMedCrossRefGoogle Scholar
  31. Rosa C, Kamita SG, Falk BW (2012) RNA interference is induced in the glassy winged sharpshooter Homalodisca vitripennis by actin dsRNA. Pest Manag Sci 68(7):995–1002 PubMedCrossRefGoogle Scholar
  32. Schröder R (2003) The genes orthodenticle and hunchback substitute for bicoid in the beetle Tribolium. Nature 422:621–625Google Scholar
  33. Tautz D, Lehmann R, Schürch H, Shuh R, Seifert E, Kienlin A, Jones K, Jäckle H (1987) Finger protein of novel structure encoded by hunchback, a second member of the gap class of Drosophila segmentation genes. Nature 327:383–389CrossRefGoogle Scholar
  34. Terenius O, Papanicolaou A, Garbutt JS, Eleftherianos I, Huvenne H, Kanginakudru S, Albrechtsen M, An C, Aymeric JL, Barthel A, Bebas P, Bitra K, Bravo A, Chevalier F, Collinge DP, Crava CM, de Maagd RA, Duvic B, Erlandson M, Faye I, Felföldi G, Fujiwara H, Futahashi R, Gandhe AS, Gatehouse HS, Gatehouse LN, Giebultowicz JM, Gómez I, Grimmelikhuijzen CJ, Groot AT, Hauser F, Heckel DG, Hegedus DD, Hrycaj S, Huang L, Hull JJ, Iatrou K, Iga M, Kanost MR, Kotwica J, Li C, Li J, Liu J, Lundmark M, Matsumoto S, Meyering-Vos M, Millichap PJ, Monteiro A, Mrinal N, Niimi T, Nowara D, Ohnishi A, Oostra V, Ozaki K, Papakonstantinou M, Popadic A, Rajam MV, Saenko S, Simpson RM, Soberón M, Strand MR, Tomita S, Toprak U, Wang P, Wee CW, Whyard S, Zhang W, Nagaraju J, Ffrench-Constant RH, Herrero S, Gordon K, Swevers L, Smagghe G (2011) RNA interference in Lepidoptera: an overview of successful and unsuccessful studies and implications for experimental design. J Insect Physiol 57:231–245PubMedCrossRefGoogle Scholar
  35. 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 bacteria expressing dsRNA of a non-midgut gene. PLoS ONE 4:1–13CrossRefGoogle Scholar
  36. Tomoyasu Y, Miller SC, Tomita S, Schoppmeier M, Grossmann D, Bucher G (2008) Exploring systemic RNA interference in insects: a genome-wide survey for RNAi genes in Tribolium. Genome Biol 9:R10PubMedCentralPubMedCrossRefGoogle Scholar
  37. 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 15:383–391PubMedCrossRefGoogle Scholar
  38. Wolff C, Sommer R, Schröder R, Glaser G, Tautz D (1995) Conserved and divergent expression aspects of the Drosophila segmentation gene hunchback in the short germband embryo of the flour beetle Tribolium. Development 121:4227–4236PubMedGoogle Scholar
  39. 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(5):e20504PubMedCentralPubMedCrossRefGoogle Scholar
  40. Zhang X, Zhang J, Zhu KY (2010) Chitosan/double-stranded RNA nanoparticle-mediated RNA interference to silence chitin synthase genes through larval feeding in the African malaria mosquito (Anopheles gambiae). Insect Mol Biol 19:683–693PubMedCrossRefGoogle Scholar
  41. Zhao YY, Liu F, Yang G, You MS (2011) PsOr1, a potential target for RNA interference-based pest management. Insect Mol Biol 20:97–104PubMedCrossRefGoogle Scholar
  42. Zhou X, Wheeler MM, Oi FM, Scharf ME (2008) RNA interference in the termite Reticulitermes flavipes through ingestion of double-stranded RNA. Insect Biochem Mol Biol 38:805–815PubMedCrossRefGoogle Scholar
  43. Zhu F, Xu J, Palli R, Ferguson J, Palli SR (2011) Ingested RNA interference for managing the populations of the Colorado potato beetle, Leptinotarsa decemlineata. Pest Manag Sci 7:175–182CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  1. 1.The Key Laboratory of Pest Management in Crops, Ministry of Agriculture, Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijingPeople’s Republic of China

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