Hostname: page-component-cd9895bd7-jn8rn Total loading time: 0 Render date: 2024-12-31T16:33:56.792Z Has data issue: false hasContentIssue false
  • English
  • Français

Midgut proteases of the cardamom shoot and capsule borer Conogethes punctiferalis (Lepidoptera: Pyralidae) and their interaction with aprotinin

Published online by Cambridge University Press:  09 March 2007

A. Josephrajkumar ,
R. Chakrabarty  and
G. Thomas
A. Josephrajkumar*
Affiliation:
Cardamom Research Station, Pampadumpara 685 556, Idukki District, Kerala, India
R. Chakrabarty
Affiliation:
Department of Plant Biology and Forest Genetics, Genetics Centre, PB no. 7080, Swedish University of Agricultural Sciences, SE 75007, Uppsala, Sweden
G. Thomas
Affiliation:
Interfield Laboratories, Kochi 682 005, Kerala, India
*
*Fax: 91 487 2370019 E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Protease inhibitors cause mortality in a range of insects, and transgenic plants expressing protease inhibitors have been protected against pest attack, particularly internal feeders that are not amenable to control by conventional means. A study of luminal proteases in Conogethes punctiferalis Guenée was performed to identify potential targets for proteinaceous biopesticides, such as protease inhibitors. The midgut protease profile of the gut lumen from C. punctiferalis was studied to determine the conditions for optimal protein hydrolysis. Optimum conditions for peptidase activity were found to be in 50 mm Tris-HCl, pH 10 containing 20 mm CaCl2; incubation for 30 min at 40°C. Four synthetic substrates, i.e. benzoyl-arg-p-nitroanilide, benzoyl-tyr-p-nitroanilide, succinyl-ala-ala-pro-leu-p-nitroanilide (SAAPLpNA) and leu-p-nitroanilide were hydrolysed by C. punctiferalis gut proteases in Tris-HCl buffer pH 10. Trypsin and elastase-like chymotrypsin were the prominent digestive proteases, and age-related modulation of midgut proteases existed for trypsin, chymotrypsin, elastase-like chymotrypsin and leucine aminopeptidase. Serine protease inhibitors such as aprotinin, soybean trypsin inhibitor and phenylmethanesulfonyl fluoride inhibited peptidase activity. Some metal ions such as Ca2+, Mg2+, Pb2+ and Co2+ enhanced BApNA-ase activity whereas others like Mn2+, Zn2+, Cu2+, Fe2+ and Hg2+ were inhibitory at 6 mm concentration. Trypsin and elastase-like chymotrypsin were significantly inhibited by 94% and 29%, respectively, by aprotinin (150 nm) under in vitro conditions. A possible incorporation of protease inhibitors into transgenic plants is discussed.

Keywords

cardamom borerConogethes punctiferalistrypsinchymotrypsinleucine aminopeptidaseaprotinin
Type
Review Article
Information
Bulletin of Entomological Research , Volume 96 , Issue 1 , February 2006 , pp. 91 - 98
Copyright
Copyright © Cambridge University Press 2006

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

Ahmad, Z., Saleemuddin, M. & Siddiqui, M. (1976) Alkaline protease in the larvae of the army worm, Spodoptera litura. Insect Biochemistry 6, 501–505.CrossRefGoogle Scholar
Ahmad, Z., Saleemuddin, M. & Siddiqui, M. (1980) Purification and characterization of three alkaline proteases from the larva of the army worm, Spodoptera litura. Insect Biochemistry 10, 667–673.CrossRefGoogle Scholar
Anwar, A. & Saleemuddin, M. (2002) Purification and characterization of a digestive alkaline protease from the larvae of Spilosoma obliqua. Archives of Insect Biochemistry and Physiology 51, 1–12.CrossRefGoogle ScholarPubMed
Applebaum, S.W. (1985) Biochemistry of digestion. pp. 279–311 in Kerkut, G.A. & Gilbert, L.I. (eds) Comprehensive insect physiology, biochemistry and pharmacology, Vol. IV. Toronto Pergamon Press.Google Scholar
Barrett, A.J. (1986) An introduction to the proteinases. pp. 3–22 in Barrett, A.J. & Salvesen, H.J. (eds) Proteinase inhibitors. Amsterdam, Elsevier.Google Scholar
Bown, D.P., Wilkinson, H.S. & Gatehouse, J.A. (1997) Differentially regulated inhibitor-sensitive and insensitive protease genes from a phytophagous insect pest, Helicoverpa armigera, are members of complex multi gene families. Insect Biochemistry and Molecular Biology 27, 625–638.CrossRefGoogle Scholar
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Analytical Biochemistry 72, 248–254.CrossRefGoogle ScholarPubMed
Burgess, E.P.J., Main, C.A., Stevens, P.S., Gatehouse, A.M.R., Christeller, J.T. & Laing, W.A. (1993) Protease inhibitors active against porina caterpillar (Wiseana cervinata). pp. 331–339 in Prestidge, R.A. (ed.) Proceedings of the Sixth Australasian Grassland Invertebrate Ecology Conference and Agricultural ResearchHamilton, New Zealand.Google Scholar
Burgess, E.P.J., Lovei, G.L., Malone, L.A., Nielsen, I.W., Gatehouse, H.S. & Christeller, J.T. (2002) Prey-mediated effects of the protease inhibitor aprotinin on the predatory carabid beetle, Nebria brevicollis. Journal of Insect Physiology 48, 1093–1101.CrossRefGoogle ScholarPubMed
Christeller, J.T., Liang, W.A., Markwick, N.P. & Burgess, E.P.J. (1992) Midgut protease activities in 12 phytophagous lepidopteran larvae: dietary and protease inhibitory interactions. Insect Biochemistry and Molecular Biology 22, 248–254.CrossRefGoogle Scholar
Christeller, J.T., Burgess, E.P.J., Mett, V., Gatehouse, H.S., Markwick, N.P., Murray, C., Malone, L.A., Wright, M.A., Philip, B.A., Watt, D., Gatehouse, L.N., Lovei, G.L., Shannon, A.L., Phung, M.M., Watson, L.M. & Laing, W.A. (2002) The expression of a mammalian proteinase inhibitor, bovine spleen trypsin inhibitor in tobacco and its effects on Helicoverpa armigera larvae. Transgenic Research 11, 161–173.CrossRefGoogle ScholarPubMed
Gatehouse, J.A., Gatehouse, A.M.R. & Brown, D.P. (2000) Control of phytophagous insect pests using serine proteinase inhibitors. pp. 9–26 in Michaud, D. Recombinant protease inhibitors in plants. Texas, USA, Landes Bioscience.Google Scholar
Gebhard, W., Tschesche, H. & Fritz, H. (1986) Biochemistry of aprotinin and aprotinin like inhibitors. pp. 375–388Barrett, A.J. & Salvesen, H.J. (eds) Proteinase inhibitors. Amsterdam Elsevier.Google Scholar
Girard, C., Le Metayer, M., Bonade-Bottino, M., Pham-Delegue, M.H. & Jouanin, L. (1998) High level of resistance to proteinase inhibitors may be conferred by proteolytic cleavage in beetle larvae. Insect Biochemistry and Molecular Biology 28, 229–237.CrossRefGoogle ScholarPubMed
Gomez, K.A. & Gomez, A.A. (1984) Statistical procedures of agricultural research. 207–215. Singapore, John Wiley and Sons.Google Scholar
Hamed, M.M.B. & Attias, J. (1987) Isolation and partial characterization of two alkaline proteases of the greater wax moth, Galleria mellonella. Insect Biochemistry 17, 653–658.CrossRefGoogle Scholar
Hilder, V.A., Gatehouse, A.M.R., Sheerman, S.E., Barker, R.F. & Boulter, D. (1987) A novel mechanism of insect resistance engineered into tobacco. Nature 30, 160–163.CrossRefGoogle Scholar
Ishaaya, I., Moore, I. & Joseph, D. (1971) Protease and amylase activity in larvae of the Egyptian cotton worm, Spodoptera littoralis. Journal of Insect Physiology 17, 945–953.CrossRefGoogle Scholar
Johnston, K.A., Lee, M.J., Gatehouse, J.A. & Anstee, J.H. (1991) The partial purification and characterization of serine protease activity in the midgut of larval Helicoverpa armigera. Insect Biochemistry 21, 389–397.CrossRefGoogle Scholar
Johnston, K.A., Lee, M.J., Brough, C., Hilder, V.A., Gatehouse, A.M.R. & Gatehouse, J.A. (1995) Protease activities in the larval midgut of Heliothis virescens: evidence for the trypsin and chymotrypsin like enzymes. Insect Biochemistry and Molecular Biology 25, 375–383.CrossRefGoogle Scholar
Kassell, B. & Laskowski, M. (1965) The basic inhibitor of bovine pancreas. V. The disulphide linkages. Biochemistry and Biophysics Research Communications 20, 463–468.CrossRefGoogle Scholar
Keller, M., Sneh, B., Strizhov, N., Prudovsky, E., Regev, A., Koncz, C., Schell, J. & Zilberstein, A. (1996) Digestion of δ-endotoxin by gut proteases may explain reduced sensitivity of advanced instar larvae of Spodoptera littoralis to CryIC. Insect Biochemistry and Molecular Biology 26, 365–373.CrossRefGoogle ScholarPubMed
Klocke, J.A. & Chan, B.G. (1982) Effects of cotton condensed tannin on feeding and digestion in the cotton pest, Heliothis zea. Journal of Insect Physiology 28, 911–915.CrossRefGoogle Scholar
Kraut, H., Frey, E.K. & Werle, E. (1930) Uber die Inaktivierung des Kallikreins. Hopp-Seyler's Z. Physiology and Chemistry 192, 1–21.CrossRefGoogle Scholar
Kumaresan, D., Regupathy, A. & Baskaran, P. (1988) Pests of spices. 10–18Nagercoil, India Rajalakshmi Publications.Google Scholar
Kunitz, M. & Northrop, J.H. (1936) Isolation from beef pancreas of crystalline trypsinogen, trypsin, trypsin inhibitor and an inhibitor trypsin compound. Journal of General Physiology 19, 991–1007.CrossRefGoogle Scholar
Lam, W., Coast, G.M. & Rayne, R.C. (1999) Isolation and characterization of two chymotrypsins from the midgut of Locusta migratoria. Insect Biochemistry and Molecular Biology 29, 653–660.CrossRefGoogle Scholar
Laskowski, M., Kato, I. Jr (1980) Protein inhibitors of proteinases. Annual Review of Biochemistry 49, 593–626.CrossRefGoogle ScholarPubMed
Lee, M.J. & Anstee, J.H. (1995) Endoproteases from the midgut of larval Spodoptera littoralis include a chymotrypsin-like enzyme with an extended binding site. Insect Biochemistry and Molecular Biology 25, 49–61.CrossRefGoogle Scholar
McManus, M.T. & Burgess, E.P.J. (1995) Effects of the soybean (Kunitz) trypsin inhibitor on growth and digestive proteases of larvae of Spodoptera litura. Journal of Insect Physiology 41, 731–738.CrossRefGoogle Scholar
McManus, M.T., White, D.W.R. & McGregor, P.G. (1994) Accumulation of a chymotrypsin inhibitor in transgenic tobacco can affect the growth of insect pests. Transgenic Research 3, 50–58.CrossRefGoogle Scholar
Metcalf, R.L. (1986) The ecology of insecticides and the chemical control of insects. pp. 251–297 in Kogan, M. (ed.) Ecological theory and integrated pest management. New York, John Wiley and Sons.Google Scholar
Novillo, C., Castanera, P. & Ortego, F. (1999) Isolation and characterization of two digestive trypsin-like proteinases from larvae of the stalk corn borer, Sesamia nonagrioides. Insect Biochemistry and Molecular Biology 29, 177–184.CrossRefGoogle ScholarPubMed
Oppert, B. (2000) Transgenic plants expressing enzyme inhibitors and the prospects for biopesticide development. pp. 83–95Koul, O. & Dhaliwal, G.S. (eds). Phytochemical biopesticides. Netherlands, Harwood Academic1.Google Scholar
Oppert, B., Hartzer, K. & Zuercher, M. (2002) Digestive proteinases in Lasioderma serricorne (Coleoptera: Anobiidae). Bulletin of Entomological Research 92, 331–336.CrossRefGoogle ScholarPubMed
Peterson, A.M., Fernando, G.J.P. & Wells, M.A. (1995) Purification, characterization and cDNA sequence of an alkaline chymotrypsin from the midgut of Manduca sexta. Insect Biochemistry and Molecular Biology 25, 765–774.CrossRefGoogle ScholarPubMed
Ravindran, P.N. & Madhusoodanan, K.J. (2002) Cardamom, the genus Elettaria. New York, Taylor & Francis Inc.Google Scholar
Samuels, R.I., Charnley, A.K. & Reynolds, S.E. (1993) A cuticle degrading proteinase from the moulting fluid of the tobacco hornworm, Manduca sexta. Insect Biochemistry and Molecular Biology 23, 607–614.CrossRefGoogle ScholarPubMed
Terra, W.R. & Ferreira, C. (1994) Insect digestive enzymes: properties compartmentalization and function. Comparative Biochemistry and Physiology 109, 1–62.Google Scholar
Terra, W.R., Ferreria, C., Jordao, B.P. & Dillon, R.J. (1996) Digestive enzymes. pp. 153–194Lehane, M.J. & Billingsley, R.F. Biology of the insect midgut. London, Chapman and Hall.Google Scholar
Thangam, E.B. & Rajkumar, G.S. (2002) Purification and characterization of alkaline protease from Alcaligenes faecalis. Biotechnology and Applied Biochemistry 35, 149–154.CrossRefGoogle ScholarPubMed
Todd, J.H., Malone, L.A., Gatehouse, H.S., Burgess, E.P.J., Christeller, J.T., Philip, B.A. & Tregidga, E.L. (2002) Effects of two protease inhibitors on larvae of Argentine stem weevil and clover root weevil. New Zealand Plant Protection 55, 416–420.CrossRefGoogle Scholar
Ussuf, K.K., Laxmi, N.H. & Mitra, R. (2001) Proteinase inhibitors: plant-derived genes of insecticidal protein for developing insect-resistant transgenic plants. Current Science 80, 847–853.Google Scholar
Varadarasan, S. (2001) Insect pest management in cardamom – key to reduce cost of production. Spice India 14, 19–22.Google Scholar
Volpicella, M., Ceci, L.R., Cordewener, J., America, T., Gallerani, R., Bode, W., Jongsma, M.A. & Beekwilder, J. (2003) Properties of purified gut trypsin from Helicoverpa zea, adapted to proteinase inhibitors. European Journal of Biochemistry 270, 10–19.CrossRefGoogle ScholarPubMed
Zhong, G.Y., Peterson, D., Delaney, D.E., Bailey, M., Witcher, D.R., Register, J.C., Bond, D., Li, C.P., Marshall, L., Kulisek, E., Ritland, D., Meyer, T., Hood, E.E. & Howard, J.A. (1999) Commercial production of aprotinin in transgenic maize seeds. Molecular Breeding 5, 345–356.CrossRefGoogle Scholar
Zhu, Y.C. & Baker, J.E. (1999) Characterization of midgut trypsin-like enzymes and three trypsinogen cDNAs from the lesser grain borer, Rhyzopertha dominica (Coleoptera: Bostrichidae). Insect Biochemistry and Molecular Biology 29, 1053–1063.CrossRefGoogle ScholarPubMed