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Carboxylamidases inSpodoptera exigua: properties and distribution in the larval body

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Abstract

Resistance to many insecticides demonstrated by the beet armyworm,Spodoptera exigua (Hübner), can be caused by the action of carboxylamidases. A colorimetric method, based on the hydrolysis of 4-nitroacetanilide to 4-nitroaniline by carboxylamidases, was used for evaluating biochemical properties of these detoxifying enzymes in beet armyworm. The optimum pH and temperature were 7.5 and 38°C, respectively. Km (Michaelis constant) and Vmax (maximal velocity) at 28°C were 2.3 X 10-4 M and 2.06 nmol min-1 mg protein-1, respectively. The enzyme activity was evaluated in several body parts and located mostly (66.2%) in the midgut. The soluble fraction (supernatant of 105,000g) contained the highest enzyme activity relative to the total (69.3%), and exhibited the highest specific activity. Carboxylamidase activity was totally inhibitedin vitro at a concentration of 10-6 M methomyl. The analysis of thein vitro inhibition kinetics indicated the ability of methomyl and diflubenzuron to inhibit carboxylamidases noncompetitively. Over 95% inhibitionin vivo was obtained when the larvae were fed with castor bean leaves dipped in 250 mgl -1 of methomyl. Thein vivo enzyme activity could be reduced to half with a pretreatment of 15 mgl -1 diflubenzuron.

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References

  1. Alava, D. and Lagunes Tejeda (1976) Resistencia cruzada a varios tipos de insecticidas despues de producir resistencia a paration metilico enSpodoptera exigua (Hübner).Folia Entomol. Mex. 36:77–78.

    Google Scholar 

  2. Aldunate, J., Repetto, Y., Letelier, M.E. and Morello, A. (1987) The carboxylesterases ofTrypanosoma cruzi epimastigotes.Comp. Biochem. Physiol. 86B:67–71.

    CAS  Google Scholar 

  3. Auda, M., El Saidy, M.F. and Degheele, D. (1988) Effect of chlorfluazuron on the cuticular protein pattern of the sixth instar larva of both susceptible and monocrotophos resistant strains ofSpodoptera littoralis (Boisd.).Meded. Fac. Landbouwwet. Rijksuniv. Gent 51:185–193.

    Google Scholar 

  4. Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Anal. Biochem. 72: 248–257.

    Article  PubMed  CAS  Google Scholar 

  5. CAB (1972) Distribution maps of pests. Pest:Spodoptera exigua (Hübner). Commonwealth Agricultural Bureaux, London. Series A, Map no. 302.

    Google Scholar 

  6. Chaufaux, J. and Ferron, P. (1986) Sensibilité de deux populations deSpodoptera exigua Hb. (Lepid., Noctuidae) aux baculovirus et aux pyretrinoïdes de synthese.Agronomie 6:99–104.

    Article  Google Scholar 

  7. Dauterman, W.C. (1985) Insect metabolism: extramicrosomal.in: Kerkut, G.A. and Gilbert, L.I. [Eds.] Comprehensive Insect Physiology, Biochemistry and Pharmacology. Vol. 12, pp. 713–730. Pergamon Press, Oxford, UK.

    Google Scholar 

  8. Dixon, M. (1953) The determination of enzyme inhibitor constants.Biochem. J. 55:170–171.

    PubMed  CAS  Google Scholar 

  9. El Saidy, M.F., Auda, M. and Degheele, D. (1989) Detoxification mechanisms of diflubenzuron and teflubenzuron in the larvae ofSpodoptera littoralis (Boisd.).Pestic. Biochem. Physiol. 35:211–222.

    Article  Google Scholar 

  10. Gazit, Y., Ishaaya, I. and Perry, A.S. (1989) Detoxification and synergism of diflubenzuron and chlorfluazuron in the red flour beetleTribolium castaneum.Pestic. Biochem. Physiol. 34:103–110.

    Article  CAS  Google Scholar 

  11. Ishaaya, I. and Degheele, D. (1988) Properties and toxicological significance of diflubenzuron hydrolase activity inSpodoptera littoralis larvae.Pestic. Biochem. Physiol. 32: 180–187.

    Article  CAS  Google Scholar 

  12. Pree, D.J., Archibald, D.E. and Cole, K.J. (1990) Insecticide resistance in spotted tentiform leafminer (Lepidoptera: Gracillariidae): Mechanisms and management.J. Econ. Entomol. 83:678–685.

    CAS  Google Scholar 

  13. Stoops, J.K. and Bender, M.L. (1975) A spectrophotometric assay for determining the rate constants of acetylcholinesterase inhibitions.Anal. Biochem. 63:543–554.

    Article  PubMed  CAS  Google Scholar 

  14. Sudderuddin, K.I. (1973) Anin vitro study of esterases, hydrolysing non-specific substrates, of an OP-resistant strain of the green peach aphid,Myzus persicae.Comp. Biochem. Physiol. 44:1067–1076.

    Article  CAS  Google Scholar 

  15. Van Asperen, K. (1962) A study of housefly esterases by means of a sensitive colorimetric method.J. Insect Physiol. 8:401–416.

    Article  CAS  Google Scholar 

  16. Van de Vrie, M. (1977)Spodoptera exigua (Lepidoptera: Noctuidae) in sierteeltgewassen.Gewasbescherming 8:67–70.

    Google Scholar 

  17. Van Laecke, K. and Degheele, D. (1991) Synergism of diflubenzuron and tefiubenzuron in larvae of beet armyworm (Lepidoptera: Noctuidae).J. Econ. Entomol. 84:785–789.

    Google Scholar 

  18. Van Laecke, K. and Degheele, D. (1991) Detoxification of diflubenzuron and tefiubenzuron in the larvae of the beet armyworm (Spodoptera exigua) (Lepidoptera: Noctuidae).Pestic. Biochem. Physiol. 40:181–190.

    Article  Google Scholar 

  19. Van Laecke, K., Degheele, D. and Auda, M. (1989) Effect of a sublethal dose of chitin synthesis inhibitors onSpodoptera exigua (Hübner) (Lepidoptera: Noctuidae).Parasitica 45:90–98.

    Google Scholar 

  20. Wilkinson, C.F. (1983) Role of mixed-function oxidases in insecticide resistance.in: Georghiou, G.P. and Saito, T. [Eds.] Pest Resistance to Pesticides. pp. 175–204. Plenum, New York, NY.

    Google Scholar 

  21. Yoshida, H.A. and Parrella, M.P. (1987) The beet armyworm in floricultural crops.Calif. Agric. 41:13–15.

    Google Scholar 

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Van Laecke, K., Degheele, D. Carboxylamidases inSpodoptera exigua: properties and distribution in the larval body. Phytoparasitica 21, 9–21 (1993). https://doi.org/10.1007/BF02980868

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