Skip to main content Accessibility help
×
Home

Identification of a novel cytochrome P450 gene, CYP321E1 from the diamondback moth, Plutella xylostella (L.) and RNA interference to evaluate its role in chlorantraniliprole resistance

  • Z. Hu (a1) (a2), Q. Lin (a1) (a2), H. Chen (a1) (a2), Z. Li (a1) (a2), F. Yin (a1) (a2) and X. Feng (a1) (a2)...

Abstract

Insect cytochrome P450 monooxygenases (P450s) play an important role in catalysis of many reactions leading to insecticides resistance. Our previous studies on transcriptome analysis of chlorantraniliprole-resistant development in the diamondback moth, Plutella xylostella revealed that up-regulation of cytochrome P450s are one of the main factors leading to the development of chlorantraniliprole resistance. Here, we report for the first time a novel cytochrome P450 gene CYP321E1, which belongs to the cytochrome P450 gene family CYP321. Real-time quantitative PCR (RT–qPCR) analyses indicated that CYP321E1 was expressed at all developmental stages of P. xylostella but was highest in the fourth-instar larvae; furthermore, the relatively high expression was observed in the midgut of the fourth-instar larvae, followed by fat bodies and epidermis. The expression of CYP321E1 in P. xylostella was differentially affected by three representative insecticides, including alphamethrin, abamectin and chlorantraniliprole. Among them, the exposure to chlorantraniliprole resulted in the largest transcript level of this cytochrome P450 gene. The findings suggested potential involvement of CYP321E1 in chlorantraniliprole resistance of P. xylostella. To assess the functional link of CYP321E1 to chlorantraniliprole resistance, RNA interference (RNAi)-mediated gene silencing by double stranded RNA (dsRNA) injecting was used. Results revealed that injection delivery of dsRNA can greatly reduce gene expression after 24 h. As a consequence of RNAi, a significant increment in mortality of larvae injected CYP321E1 dsRNA was observed after 24 h of exposure to chlorantraniliprole. These results strongly support our notion that this novel cytochrome P450 gene plays an important role in chlorantraniliprole detoxification in the diamondback moth and is partly responsible for its resistance.

  • View HTML
    • Send article to Kindle

      To send this article to your Kindle, first ensure no-reply@cambridge.org is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about sending to your Kindle. Find out more about sending to your Kindle.

      Note you can select to send to either the @free.kindle.com or @kindle.com variations. ‘@free.kindle.com’ emails are free but can only be sent to your device when it is connected to wi-fi. ‘@kindle.com’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

      Find out more about the Kindle Personal Document Service.

      Identification of a novel cytochrome P450 gene, CYP321E1 from the diamondback moth, Plutella xylostella (L.) and RNA interference to evaluate its role in chlorantraniliprole resistance
      Available formats
      ×

      Send article to Dropbox

      To send this article to your Dropbox account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Dropbox.

      Identification of a novel cytochrome P450 gene, CYP321E1 from the diamondback moth, Plutella xylostella (L.) and RNA interference to evaluate its role in chlorantraniliprole resistance
      Available formats
      ×

      Send article to Google Drive

      To send this article to your Google Drive account, please select one or more formats and confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your <service> account. Find out more about sending content to Google Drive.

      Identification of a novel cytochrome P450 gene, CYP321E1 from the diamondback moth, Plutella xylostella (L.) and RNA interference to evaluate its role in chlorantraniliprole resistance
      Available formats
      ×

Copyright

The online version of this article is published within an Open Access environment subject to the conditions of the Creative Commons Attribution licence http://creativecommons.org/licenses/by/3.0/

Corresponding author

* Author for correspondence Phone & Fax: +86-20-87597577 E-mail: fengx@gdppri.com

References

Hide All
Bautista, M.A.M., Tanaka, T. & Miyata, T. (2007) Identification of permethrin-inducible cytochrome P450s from the diamondback moth, Plutella xylostella (L) and the possibility of involvement in permethrin resistance. Pesticide Biochemistry and Physiology 87, 8593.
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, 3846.
Baek, J.H., Clark, J.M. & Lee, S.H. (2010) Cross-strain comparison of cypermethrin-induced cytochrome P450 transcription under different induction conditions in diamondback moth. Pesticide Biochemistry and Physiology 96, 4350.
Brandt, A., Scharf, M., Pedra, J.H.F., Holmes, G., Dean, A., Kreitman, M. & Pittendrigh, B.R. (2002) Different expression and induction of two Drosophila cytochrome P450 genes near the Rst(2) DDT locus. Insect Molecular Biology 11, 337341.
Brun, A., Cuany, A., Le Mouel, T., Bere, 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, 697703.
Chung, H., Sztal, T., Pasricha, S., Sridhar, M., Batterham, P. & Daborn, P.J. (2009) Characterization of Drosophila melanogaster cytochrome P450 genes. Proceedings of the National Academy of Science of the United States of America 89, 1092010924.
Feyereisen, R. (1999) Insect P450 enzymes. Annual Review of Entomology 44, 507533.
Feyereisen, R. (2005) Insect cytochrome P450. Vol. 4, pp. 177 in Gilbert, L.I., Iatrou, K. & Gill, S.S. (Eds) Comprehensive Molecular Insect Science. Oxford, Elsevier.
Feyereisen, R. (2006) Evolution of insect P450. Biochemical Society Transactions 34, 12521255.
Feyereisen, R. (2012) Insect CYP genes and P450 enzymes. pp. 236316 in Gilbert, L. (Ed) In Insect Molecular Biology and Biochemistry. London, Elsevier.
Fuchs, S.Y., Spiegelman, V.S. & Belitsky, G.A. (1994) Inducibility of various cytochrome-P450 isozymes by phenobarbital and some other xenobiotics in Drosophila melanogaster . Biochemical Pharmacology 47, 18671873.
Furlong, M.J., Wright, D.J. & Dosdall, L.M. (2013) Diamondback moth ecology and management: problems, progress, and prospects. Annual Review of Entomology 58, 517541.
Guo, Y., Zhang, J., Yu, R., Zhu, K.Y., Guo, Y. & Ma, E. (2012) Identification of two new cytochrome P450 genes and RNA interference to evaluate their roles in detoxification of commonly used insecticides in Locusta migratoria . Chemosphere 87, 709717.
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, 179189.
Hodgson, E. (1985) Microsomal monooxygenases. pp. 647721 in Kerkut, G.A. & Gilbert, L.I. (Eds) Comprehensive Insect Physiology, Biochemistry, and Pharmacology. Oxford, Pergamon.
Le Goff, G., Hilliou, F., Siegfried, G.D., Boundy, S., Wajnberg, E., Sofer, L., Audant, P., ffrench-Constant, R.H. & Feyereisen, R. (2006) Xenobiotic response in Drosophila melanogaster: sex dependence of P450 and GST gene induction. Insect Biochemistry and Molecular Biology 36, 674682.
Li, A., Yang, Y., Wu, S., Li, C. & Wu, Y. (2006) Investigation on the resistance mechanisms to fipronil in diamondback moth. Journal of Economic Entomology 99, 914919.
Li, X., Schuler, M.A. & Berenbaum, M.R. (2007) Molecular mechanisms of metabolic resistance to synthetic and natural xenobiotics. Annual Review of Entomology 52, 231253.
Lin, Q.S., Jin, F.L., Hu, Z.D., Chen, H.Y., Yin, F., Li, Z.Y., Dong, X.L., Zhang, D.Y., Ren, S.X. & Feng, X. (2013) Transcriptome analysis of chlorantraniliprole resistance development in the diamondback moth Plutella xylostella . PLoS ONE 8(8), e72314.
Liu, N. & Scott, J.G. (1996) Genetic analysis of factors controlling elevated cytochrome P450, CYP6D1, cytochrome b5, P450 reductase and monooxygenase activities in LPR house flies, Musca domestica . Biochemical Genetics 34, 133148.
Liu, N. & Scott, J.G. (1998) Increased transcription of CYP6D1 causes cytochrome P450-mediated insecticide resistance in housefly. Insect Molecular Biology 28, 531535.
Nelson, D.R. (2009) The cytochrome P450 homepage. Human Genomics 4, 5965.
Ninsin, K.D. & Tanaka, T. (2005) Synergism and stability of acetamiprid resistance in a laboratory colony of Plutella xylostella . Pest Management Science 61, 723727.
Poupardin, R., Reynaud, S., Strode, C., Ranson, H., Vontas, J. & David, J.P. (2008) Cross- induction of detoxification genes by environmental xenobiotics and insecticides in the mosquito Aedes aegypti: impact on larval tolerance to chemical insecticides. Insect Biochemistry and Molecular Biology 38, 540551.
Ranasinghe, C. & Hobbs, A.A. (1998) Isolation and characterization of two cytochrome P450 cDNA clones for CYP6B6 and CYP6B7 from Helicoverpa armigera (Hubner): possible involvement of CYP6B7 in pyrethroid resistance. Insect Biochemistry and Molecular Biology 28, 571580.
Scott, J.G. & Wen, Z. (2001) Cytochromes P450 of insects: the tip of the iceberg. Pest Management Science 57, 958967.
Shen, B., Dong, H.Q., Tian, H.S., Ma, L., Li, X.L., Wu, G.L. & Zhu, C.L. (2003) Cytochrome P450 genes expressed in the deltamethrin-susceptible and – resistant strains of Culex pipiens pallens . Pesticide Biochemistry and Physiology 75, 1926.
Snyder, M.J., Stevens, J.L., Andersen, J.F. & Feyereisen, R. (1995) Expression of cytochrome P450 genes of the CYP4 family in midgut and fat body of the tobacco hornworm, Manduca sexta . Archives of Biochemistry and Biophysics 321, 1320.
Tabashnik, B.E., Cushing, N.L. & Finson, N. (1987) Leaf residue vs topical bioassay for assessing resistance in the diamondback moth (Lepidoptera: Plutellidae). FAO Plant Protection Bulletin 35, 1114.
Talekar, N.S. & Shelton, A.M. (1993) Biology, ecology and management of diamondback moth. Annual Review of Entomology 38, 275301.
Troczka, B., Zimmer, C.T., Elias, J., Schorn, C., Bass, C., Davies, T.G.E., Field, L.M., Williamson, M.S., Slater, R. & Nauen, R. (2012) Resistance to diamide insecticides in diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae) is associated with a mutation in the membrane-spanning domain of the ryanodine receptor. Insect Biochemistry and Molecular Biology 42, 873880.
Wang, X.L. & Wu, Y.D. (2012) High levels of resistance to chlorantraniliprole evolved in field populations of Plutella xylostella . Journal of Economic Entomology 105, 10191023.
Wang, X.L., Li, X.Y. & Wu, Y.D. (2010) Baseline susceptibility of the diamondback moth (Lepidoptera: Plutellidae) to chlorantraniliprole in China. Journal of Economic Entomology 103, 843848.
Wang, X.L., Khakame, S.K., Ye, C., Yang, Y.H. & Wu, Y.D. (2013) Characterisation of field-evolved resistance to chlorantraniliprole in the diamondback moth, Plutella xylostella, from China. Pest Management Science 69(5), 661665.
Zhou, X.J., Ma, C.X., Li, M., Sheng, C.F., Liu, H.X. & Qiu, X.H. (2010) CYP9A12 and CYP9A17 in the cotton bollworm Helicoverp armigera: sequence similarity, expression profile and xenobiotic response. Pest Management Science 66, 6573.

Keywords

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed