Hostname: page-component-cd9895bd7-jkksz Total loading time: 0 Render date: 2024-12-18T18:54:16.190Z Has data issue: false hasContentIssue false
  • English
  • Français

Silencing of cytochrome P450 CYP6B6 gene of cotton bollworm (Helicoverpa armigera) by RNAi

Published online by Cambridge University Press:  16 April 2013

X. Zhang ,
X. Liu ,
J. Ma  and
J. Zhao
X. Zhang
Affiliation:
College of Life Science and Technology, Xinjiang University, Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, Urumqi 830046, China
X. Liu*
Affiliation:
College of Life Science and Technology, Xinjiang University, Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, Urumqi 830046, China
J. Ma
Affiliation:
College of Life Science and Technology, Xinjiang University, Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, Urumqi 830046, China
J. Zhao
Affiliation:
College of Life Science and Technology, Xinjiang University, Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, Urumqi 830046, China
*
*Author for correspondence Phone: (+86)9918582076 E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

RNA interference (RNAi) induced through double-stranded RNA (dsRNA) has been used widely to study gene function in insects. In this paper we demonstrate the efficacy of RNAi in the cotton bollworm, Helicoverpa armigera. Using CYP6B6 as the target gene, which is expressed in the fat baby and midgut of the lepidopteran pest H. armigera, we constructed the vector which expressed dsRNA of CYP6B6. Northern blot analysis showed that dsRNA expressed in the Escherichia coli (HT115) was target gene. The results also showed that the gene expression level and protein expression level of H. armigera larvae fed with dsRNA expressed by E. coli were significantly lower than those of all controls, but the gene expression level was more obvious than that at the protein level; significant lethality differences were also found between HT115 bacteria containing L4440-dsC1 treatment and HT115 bacteria containing L4440 vector or CK (ddH2O) in instar larvae on 4 day when continuous feeding, 32.45% mortality was recorded in the group of feeding HT115 bacteria containing L4440-dsC1 on 10 day. Our results suggest that the RNAi pathway can be exploited to control insect pests.

Keywords

RNA interferenceHelicoverpa armigeraCYP6B6double-stranded RNA
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 103 , Issue 5 , October 2013 , pp. 584 - 591
Copyright
Copyright © Cambridge University Press 2013 

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

Alves, A.P., Lorenzen, M.D., Beeman, R.W., Foster, J.E. & Siegfried, B.D. (2010) RNA interference as a method for target-site screening in the western corn rootworm, Diabrotica virgifera virgifera. Journal of Insect Science 10, 162.Google Scholar
Amdam, G., Simões, Z., Guidugli, K., Norberg, K. & Omholt, S. (2003) Disruption of vitellogenin gene function in adult honeybees by intra-abdominal injection of double-stranded RNA. BMC Biotechnology 3, 1–8.Google Scholar
Araujo, R., Santos, A., Pinto, F., Gontijo, N., Lehane, M. & Pereira, M. (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.Google Scholar
Bautista, M.A.M., Miyata, T., Miura, K. & Tanaka, T. (2009) RNA interference-mediated knockdown of a cytochrome P450, CYP6BG1, from the diamondback moth, Plutella xylostella, reduces larval resistance to permethrin. Insect Biochemistry and Molecular Biology 39, 38–46.Google Scholar
Brun, A., Cuany, A., Le Mouel, T., Berge, J. & Amichot, M. (1996) Inducibility of the Drosophila melanogaster cytochrome P450 gene, CYP6A2, by phenobarbital in insecticide susceptible or resistant strains. Insect Biochemistry and Molecular Biology 26, 697–703.CrossRefGoogle ScholarPubMed
Bucher, G., Scholten, J. & Klingler, M. (2002) Parental RNAi in Tribolium (Coleoptera). Current Biology 12, 85–86.CrossRefGoogle ScholarPubMed
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 interference. Bulletin of Entomological Research 98, 613–619.Google Scholar
Dong, Y. & Friedrich, M. (2005) Nymphal RNAi: systemic RNAi mediated gene knockdown in juvenile grasshopper. BMC Biotechnology 5, 25.Google Scholar
Eleftherianos, I., Millichap, P.J. & Reynolds, S.E. (2006) 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
Enayati, A., Ranson, H. & Hemingway, J. (2005) Insect glutathione transferases and insecticide resistance. Insect Molecular Biology 14, 3–8.Google Scholar
Feyereisen, R. (2005) Insect cytochrome P450. Comprehensive Molecular Insect Science 4, 1–77.Google Scholar
Fire, A., Xu, S.Q., Montgomery, M.K., Kostas, S.A., Driver, S.E. & Mello, C.C. (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806–811.Google Scholar
Grubor, V.D. & Heckel, D.G. (2007) Evaluation of the role of CYP6B cytochrome P450s in pyrethroid resistant Australian Helicoverpa armigera. Insect Molecular Biology 16, 15–23.Google Scholar
Harrison, T.L., Zangerl, A.R., Schuler, M.A. & Berenbaum, M.R. (2001) Developmental variation in cytochrome P450 expression in Papilio polyxenes in response to xanthotoxin, a host plant allelochemical. Archives of Insect Biochemistry and Physiology 48, 179–189.Google Scholar
Hemingway, J. & Karunaratne, S.H. (1998) Mosquito carboxylesterases: a review of the molecular biology and biochemistry of a major insecticide resistance mechanism. Medical and Veterinary Entomology 12, 1–12.Google Scholar
Hlavica, P. (2011) Insect cytochromes P450: topology of structural elements predicted to govern catalytic versatility. Journal of Inorganic Biochemistry 105, 1354–1364.Google Scholar
Hodgson, E. (1985) Microsomal monooxygenases. pp. 647–712in Kerkut, G.A. & Gilbert, L.I. (Eds) Comprehensive Insect Physiology, Biochemistry, and Pharmacology. Oxford, Pergamon.Google Scholar
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.Google Scholar
Jaubert-Possamai, S., Trionnaire, G.L., Bonhomme, J., Christophides, G., Rispe, C. & Tagu, D. (2007) Gene knockdown by RNAi in the pea aphid Acyrthosiphon pisum. BMC Biotechnology 7, 63.Google Scholar
Le Goff, G., Hilliou, F., Siegfried, B.D., Boundy, S., Wajnberg, E., Sofer, L., Audant, P. & Feyereisen, R. (2006) Xenobiotic response in Drosophila melanogaster: sex dependence of P450 and GST gene induction. Insect Biochemistry and Molecular Biology 36, 674–682.Google Scholar
Liu, X., Liang, P., Gao, X. & Shi, X. (2006) Induction of the cytochrome P450 activity by plant allelochemicals in the cotton bollworm, (Helicoverpa armigera) (Hübner). Pesticide Biochemistry and Physiology 84, 127–134.Google Scholar
Mao, Y., Cai, W., Wang, J., Hong, G., Tao, X., Wang, L., Huang, Y. & Chen, X. (2007) Silencing a cotton bollworm P450 monooxygenase gene by plant-mediated RNAi impairs larval tolerance of gossypol. Nature Biotechnology 25, 1307–1313.Google Scholar
Newmark, P.A., Reddien, P.W., Cebrià, F. & Alvarado, A.S. (2003) Ingestion of bacterially expressed double-stranded RNA inhibits gene expression in planarians. Proceedings of the National Academy of Sciences of the United States of America 100, 11861–11865.Google Scholar
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.Google Scholar
Rajagopal, R., Sivakumar, S., Agrawal, N., Malhotra, P. & Bhatnagar, R.K. (2002) Silencing of midgut aminopeptidase N of Spodoptera litura by double-stranded RNA establishes its role as Bacillus thuringiensis toxin receptor. Journal of Biological Chemistry 277, 46849–46851.Google Scholar
Scott, J.G. (1999) Cytochromes P450 and insecticide resistance. Insect Biochemistry and Molecular Biology 29, 757–777.CrossRefGoogle ScholarPubMed
Solis, C.F., Santi-Rocca, J., Perdomo, D., Weber, C. & Guillén, N. (2009) Use of bacterially expressed dsRNA to downregulate Entamoeba histolytica gene expression. PLoS ONE 4, e8424.Google Scholar
Stegeman, J.J. & Livingstone, D.R. (1998) Forms and functions of cytochrome P450. Comparative Biochemistry and Physiology. Part C, Pharmacology, Toxicology & Endocrinology 121, 1–3.Google Scholar
Terra, W., Ferreira, C. & Baker, J. (1996) Compartmentalization of digestion. pp. 206–235in Lehane, M.J. & Billingsley, P.F. (Eds) Biology of the Insect Midgut. London, Chapman & Hall.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 bacteria expressing dsRNA of a non-midgut gene. PLoS ONE 4, e6225.Google Scholar
Timmons, L. & Fire, A. (1998) Specific interference by ingested dsRNA. Nature 395, 854.Google Scholar
Timmons, L. & Fire, A. (2001) Ingestion of bacterially expressed dsRNAs can produce specific and potent genetic interference in (Caenorhabditis elegans). Gene 263, 103–112.Google Scholar
Tomoyasu, Y. & Denell, R.E. (2004) Larval RNAi in Tribolium (Coleoptera) for analyzing adult development. Development Genes and Evolution 214, 575–578.Google Scholar
Turner, C., Davy, M., MacDiarmid, R., Plummer, K., Birch, N. & Newcomb, R. (2006) RNA interference in the light brown apple moth, Epiphyas postvittana (Walker) induced by double-stranded RNA feeding. Insect Molecular Biology 15, 383–391.Google Scholar
Walshe, D., Lehane, S., Lehane, M. & Haines, L. (2009) Prolonged gene knockdown in the tsetse fly Glossina by feeding double stranded RNA. Insect Molecular Biology 18, 11–19.Google Scholar
Willoughby, L., Chung, H., Lumb, C., Robin, C., Batterham, P. & Daborn, P.J. (2006) A comparison of Drosophila melanogaster detoxification gene induction responses for six insecticides, caffeine and phenobarbital. Insect Biochemistry and Molecular Biology 36, 934–942.Google Scholar