Eur. J. Entomol. 110 (1): 39-45, 2013 | 10.14411/eje.2013.005

Expression of cytochrome P450 CYP6B6 in the different developmental stages of the insect Helicoverpa armigera (Lepidoptera: Noctuidae)

Xuetao ZHANG, Dan YUAN, Li DING, Pan LI, Fen LI, Xiaoning LIU*
College of Life Science and Technology, Xinjiang University, Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, Urumqi 830046, China; e-mails: zhxt51@163.com; 79546571haha@sina.com; 2012dingli@sina.com; lipan19860706@yahoo.com.cn; lifen2010happy@sina.com; liuxn0103@sina.com
* Corresponding author.

In order to determine when during the development of H. armigera CYP6B6 is expressed, we measured the width of the head capsule, the length of the body and weight of the different life stages of the cotton bollworm reared under laboratory condition. The larvae took about 20 d to complete their development under these conditions, the pre-pupae about 4 d, pupae about 9 d, and the adults lived for about 9 d and the eggs took about 4 d to hatch. Knowing this it was possible to determine the pattern of cytochrome P450 CYP6B6 expression at different stages in the life cycle of H. armigera using real-time quantitative polymerase chain reaction (RT-qPCR). The CYP6B6 gene was expressed in all the developmental stages of the cotton bollworm. The results indicate that the level of expression was lower in the egg, pre-pupal, pupal and adult stages than in larvae. The level of expression of CYP6B6 decreased from the 1st to the 3rd instar larva, with the lowest level recorded in the 3rd instar larvae and then increased reaching its highest value in 6th instar larvae. A similar trend in the expression of CYP6B6 was detected using immunohistochemistry. As the results show, more efficient control of the cotton bollworm can be achieved if insecticides are applied when the larvae of H. armiger are in the 3rd instar, because at that stage the eat little food and there is a low level of detoxifying enzymes in the larvae.

Keywords: Lepidoptera, Noctuidae, Helicoverpa armigera, developmental stage, CYP6B6 expression

Received: April 11, 2012; Accepted: June 22, 2012; Published: January 2, 2013

Download citation

References

  1. AHMAD M., GLADWELL R.T. & MCCAFFERY A.R. 1989: Decreased nerve sensitivity is a mechanism of resistance in a pyrethroid resistant strain of Heliothis armigera from Thailand. - Pestic. Biochem. Physiol. 35: 165-171 Go to original source...
  2. BRATTSTEN L.B., HOLYOKE J.C.W., LEEPER J.R. & RAFFA K.F. 1986: Insecticide resistance: Challenge to pest management and basic research. - Science 231: 1255-1260 Go to original source...
  3. CHEN S., YANG Y. & WU Y. 2005: Correlation between fenvalerate resistance and cytochrome P450-mediated O-demethylation activity in Helicoverpa armigera (Lepidoptera: Noctuidae). - J. Econ. Entomol. 98: 943-946 Go to original source...
  4. FEYREISEN R. 2005: Insect cytochrome P450. In: Gilbert L.I., Iatrou K. & Gill S.S. (eds): Comprehensive Molecular Insect Science - Biochemistry and Molecular Biology. Vol. 4. Elsevier, Oxford, pp. 1-77
  5. FORRESTER N.W., CAHILL M., BIRD L.J. & LAYLAND J.K. 1993: Management of pyrethroid and endosulfan resistance in Helicoverpa armigera (Lepidoptera: Noctuidae) in Australia. - Bull. Entomol. Res. 1: 1-132 Go to original source...
  6. GANDHI R., VARAK E. & GOLDBERG M.L. 1992: Molecular analysis of a cytochrome P450 gene of family 4 on the Drosophila X chchromosome. - DNA Cell Biol. 11: 397-404 Go to original source...
  7. GUNNING R.V., EASTON C.S., BALFE M.E. & FERRIS I.G. 1991: Pyrethroid resistance mechanisms in Australian Helicoverpa armigera. - Pestic. Sci. 33: 473-490 Go to original source...
  8. 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 allelochemicals. - Arch. Insect Biochem. Physiol. 48: 179-189 Go to original source...
  9. HELVIG C., KOENER G., UNNITHAN G.C. & FEYREISEN R. 2004: CYP15A1, the cytochrome P450 that catalyzes epoxidation of methyl farnesoate to juvenile hormone III in cockroach corpora allata. - Proc. Natl Acad. Sci. USA. 101: 4024-4029 Go to original source...
  10. HODGSON E. 1983: The significance of cytochrome P450 in insects. - Insect Biochem. 13: 237-246 Go to original source...
  11. HODGSON E., ROSE R.L., RYU D.Y., FALLS G., BLAKE B.L. & LEVI P.E. 1995: Pesticide-metabolizing enzymes. - Toxicol. Lett. 3: 73-81 Go to original source...
  12. HUNTER M.D. & MCNEIL J.N. 1997: Host-plant quality influences diapause and voltinism in a polyphagous insect herbivore. - Ecology 78: 977-986 Go to original source...
  13. KASAI S. & TOMITA T. 2003: Male specific expression of a cytochrome P450 (Cyp312a1) in Drosophila melanogaster. - Biochem. Biophys. Res. Commun. 300: 894-900 Go to original source...
  14. KORYTKO P.J. & SCOTT J.G. 1998: CYP6D1 protects thoracic ganglia of houseflies from the neurotoxic insecticide cypermethrin. - Arch. Insect Biochem. Physiol. 37: 57-63 Go to original source...
  15. LI X., BERENBAUM M.R. & SCHULER M.A. 2002: Plant allelochemicals differentially regulate Helicoverpa zea cytochrome P450 genes. - Insect Mol. Biol. 11: 343-351 Go to original source...
  16. LIU Z.D., LI D.M., GONG P.Y. & WU K.J. 2004: Life table studies of the cotton bollworm, Helicoverpa armigera (Huebner) (Lepidoptera: Noctuidae), on different host plants. - Environ. Entomol. 33: 1570-1576 Go to original source...
  17. LIU X.N., LIANG P., GAO X.W. & SHI X. 2006: Induction of the cytochrome P450 activity by allelochemicals in the cotton bollworm, Helicoverpa armigera (Huebner). - Pestic. Biochem. Physiol. 84: 127-134 Go to original source...
  18. JIANG M.X. & ZHANG X.X. 1998: Effects of photoperiods on developmental time and pupal weight of the cotton bollworm, Helicoverpa armigera. - Entomol. Knowl. 35: 71-73
  19. MCCAFFERY A.R., HEAD D., TAN J., DUBBLEDAM A.A., SUBRANMARIAM V.R. & AGHAN A. 1997: Nerve insensitivity resistance to pyrethroid in heliothine Lepidoptera. - Pestic. Sci. 44: 237-247 Go to original source...
  20. MAIBECHE-COISINE M., MONTI-DEDIEU L., ARAGON S. & DAUPHINVILLEMANT C. 2000: A new cytochrome P450 from Drosophila melanogaster, CYP4G15, expressed in the nervous system. - Biochem. Biophys. Res. Commun. 273: 1132-1137 Go to original source...
  21. NIKOU D., RANSON H. & HEMINGWAY J. 2003: An adult-specific CYP6 P450 gene is overexpressed in a pyrethroid-resistant strain of the malaria vector, Anopheles gambiae. - Gene 318: 91-102 Go to original source...
  22. QIU X.H. & LENG X.F. 2000: The tissue distribution of monooxygenase activities of cotton bollworm, Helicoverpa armigera. - Acta Ecol. Sin. 20: 299-303
  23. QIU L.H., ZHANG W.J. & LI X.W. 1999: Changes of component contents and enzyme activities of microdomal mixed function oxidases in Helicoverpa armigera (Hubner) during different stages. - Chin. J. Pestic. Sci. 1: 45-50 [in Chinese, English abstr]
  24. QIU X.H., LI W. & LENG X.F. 2001: Comparison of components and activities of microsomal P450 monooxygenases from the cotton bollworm, Helicoverpa armigera during different developmental stages. - Acta Entomol. Sin. 44: 142-147
  25. QIU X.H., LI W., TIAN Y. & LENG X.F. 2003: Cytochrome P450 monooxygenases in the cotton bollworm (Lepidoptera: Noctuidae): tissue difference and induction. - J. Econ. Entomol. 4: 1283-1289 Go to original source...
  26. RANASINGHE C. & HOBBS A.A. 1998: Isolation and characterization of two cytochrome P450 cDNA clones for CYP6B6 and CYP6B7 from Helicoverpa armigera (Huebner): possible involvement of CYP6B7 in pyrethroid resistance. - Insect Biochem. Mol. Biol. 28: 571-580 Go to original source...
  27. RANASINGHE C., HEADLAM M. & HOBBS A.A. 1997: Induction of the mRNA for CYP6B2, a pyrethroid inducible cytochrome P450, in Helicoverpa armigera (Hubner) by dietary monoterpenes. - Arch. Insect Biochem. Physiol. 34: 99-109 Go to original source...
  28. SCOTT J.G., LIU N & WEN Z.M. 1998: Insect cytochromes P450: diversity, insecticide resistance and tolerance to plant toxins. - Comp. Biochem. Physiol. (C) 121: 147-155
  29. SUTHERLAND T.D., UNNITHAN G.C., ANDERSEN J.F., EVANS P.H., MURATALIEV M.B., SZABO L.Z., MASH E.A., BOWERS W.S. & FEYREISEN R. 1998: A cytochrome P450 terpenoid hydroxylase linked to the suppression of insect juvenile hormone synthesis. - Proc. Natl Acad. Sci. USA 95: 12884-12889 Go to original source...
  30. TAN W. & GUO Y. 1996: Effects of host plant on susceptibility to deltamethrin and detoxification enzyme of Heliothis armigera (Lepidoptera: Noctuidea). - J. Econ. Entomol. 89: 11-14 Go to original source...
  31. TAYLOR R.L. 1931: On "Dyar's rule" and its application to sawfly larvae. - Ann. Entomol. Soc. Am. 24: 451-466 Go to original source...
  32. WEN Z. & SCOTT J.G. 2001: Cytochrome P450 CYP6L1 is specifically expressed in the reproductive tissues of adult male German cockroaches, Blattella germanica (L.). - Insect Biochem. Mol. Biol. 31: 179-187 Go to original source...
  33. WU Y.C. 1977: A survey on the peanut noctuids in Kwangtung province. - Acta Entomol. Sin. 4: 445-450 [in Chinese, English abstr]
  34. WU K. & GONG P. 1997: A new and practical artificial diet for the cotton bollworm. - Insect Sci. 4: 277-282 Go to original source...
  35. WU K.J. & LI M.H. 1993: Nutritional ecology of the cotton bollworm, Helicoverpa armigera (Huebner): life tables of the population on the artificial diets whit different protein levels. - Acta Entoml. Sin. 36: 21-28 [in Chinese, English abstr]
  36. WILLOUGHBY L., CHUNG H., LUMB C., ROBIN C., BATTERHAM P. & DARBON P.J. 2006: A comparison of Drosophila melanogaster detoxification gene induction responses for six insecticides, caffeine and phenobarbital. - Insect Biochem. Mol. Biol. 36: 934-942 Go to original source...
  37. YANG Y., WU Y., CHENA S., DEVINEB G.J., DENHOLM I., JEWESS P. & MOORES G.D. 2004: The involvement of microsomal oxidases in pyrethroid resistance in Helicoverpa armigera from Asia. - Insect Biochem. Mol. Biol. 34: 763-773 Go to original source...