Accumulation of a chymotrypsin inhibitor in transgenic tobacco can affect the growth of insect pests
Tóm tắt
A member of the potato proteinase inhibitor II (PPI II) gene family that encodes for a chymotrypsin iso-inhibitor has been introduced into tobacco (Nicotiana tabacum) usingAgrobacterium tumefaciens-mediated T-DNA transfer. Analysis of the primary transgenic plants (designated R0) confirmed that the introduced gene is being expressed and the inhibitor accumulates as an intact and fully functional protein. For insect feeding trials, progeny from the self-fertilization of R0 plants (designated R1) were used. Leaf tissue, either from transgenic or from control (non-transgenic) plants, was fed to larvae ofChrysodeixis eriosoma (Lepidoptera: Noctuidae, green looper),Spodoptera litura (F.) (Lepidoptera: Noctuidae) andThysanoplusia orichalcea (F.) (Lepidoptera: Noctuidae) and insect weight gain (increase in fresh weight) measured. Consistently,C. eriosoma larvae fed leaf tissue from transgenic plants expressing thePPI II gene grew slower than insects fed leaf tissue from non-transgenic plants or transgenic plants with no detectablePPI II protein accumulation. However, larvae of bothS. litura andT. orichalcea consistently demonstrated similar or faster growth when fed leaf tissue from transgenic plants compared with those fed non-transgenic plants. In agreement with the feeding trials, the chymotrypsin iso-inhibitor extracted from transgenic tobacco effectively retarded chymotrypsin-like activity measured inC. eriosoma digestive tract extracts, but not in extracts fromS. litura. We conclude, therefore, that for certain insects the use of chymotrypsin inhibitors should now be evaluated as an effective strategy to provide field resistance against insect pests in transgenic plants, but further, that a single proteinase inhibitor gene may not be universally effective against a range of insect pests. The significance of these observations is discussed with respect to the inclusion of chymotrypsin inhibitors in the composite of insect pest resistance factors that have been proposed for introduction into crop plants.
Tài liệu tham khảo
Bevan, M. (1984) BinaryAgrobacterium vectors for plant transformation.Nuc. Acids Res. 12, 8711–21.
Boulter, D., Edwards, G.A., Gatehouse, A.M.R., Gatehouse, J.A. and Hilder, V.A. (1990) Additive protective effects of different plant-derived insect resistance genes in transgenic tobacco plants.Crop Protect. 9, 351–4.
Broadway, R.M. and Duffy, S.S. (1986) Plant proteinase inhibitors: Mechanism of action and effect on the growth and digestive physiology of larvalHeliothis zea andSpodoptera exigua.J. Insect Physiol. 32, 827–33.
Bryant, J., Green, T.R., Gurusaddaiah, T. and Ryan, C.A. (1976) Proteinase inhibitor II from potatoes: Isolation and characterization of its protomar components.Biochem. 15, 3418–24.
Christeller, J.T., Shaw, B.D., Gardiner, S.E. and Dymock, J. (1989) Partial purification and characterisation of the major midgut proteases of grassgrub larvae (Costelytra zelandicae, Coleoptera: Scarabaeidae).Insect Biochem. 19, 221–31.
Ditta, G., Stanfield, S., Corbins, D. and Helinski, D.R. (1980) Broad host range DNA cloning system for Gram-negative bacteria: construction of a gene bank ofRhizobium meliloti.Proc. Natl Acad. Sci. USA 77, 7347–51.
Garcia-Olmedo, F., Salcedo, G., Sanchez-Monge, R., Gomez, L., Royo, J. and Carbonero, P. (1987) Plant proteinaceous inhibitors of proteinases and α-amylases.Oxf. Surv. Pl. Cell Mol. Biol. 4, 275–334.
Gatehouse, A.M.R. and Boulter, D. (1983) Assessment of the antimetabolic effects of trypsin inhibitors from cowpea (Vigna unguiculata) and other legumes on development of the bruchid beetleCallosobruchus maculatus.J. Sci. Food Agric. 34, 345–50.
Hernalsteens, J.P., Vliet, F. van, Beuckeleer, M. De, Depicker, A., Engler, G., Lemmers, M., Hosters, M., Montagu, M. van and Schell, J. (1980) TheAgrobacterium tumefaciens Ti plasmid as a host vector system for introducing foreign DNA into plant cells.Nature 287, 654–6.
Herrera-Estrella, L., Depicher, A., Montagu, M. van and Schell, J. (1983) Expression of chimaeric genes transferred into plant cells using a Ti-plasmid-derived vector.Nature 303, 209–13.
Hilder, V.A., Gatehouse, A.M.R., Sheerman, S.E., Barker, R.F. and Boulter, D. (1987) A novel mechanism of insect resistance engineered into tobacco.Nature 330, 160–3.
Hill, R.L. (1989)Spodoptera litura (F.), tropical armyworm (Lepidoptera: Noctuidae). InA review of biological control of insect pests and weeds in New Zealand 1874–1987, pp. 143–4 (eds P.G. Cameron, R.L. Hill, J. Bain and W.P. Thomas). Wallingford: CAB International.
Hill, M.G., Cameron, P.G., Dugdale, J.S., Allan, D.J. and Walker, G.P. (1987) Biology ofThysanoplusia orichalcea (Lepidoptera: Noctuidae) in New Zealand.N. Z. Entomol. 10, 44–50.
Horsch, R.B., Fry, J.E., Hoffman, N.L., Eichholtz, D., Rogers, S.G. and Fraley, R.F. (1985) A simple and general method for transferring genes into plants.Science 227, 1299–31.
Johnson, R., Narvaez, J., An, G. and Ryan, C. (1990) Expression of proteinase inhibitors I and II in transgenic tobacco plants: Effects on natural defense againstManduca sexta larvae.Proc. Natl Acad. Sci. USA 86, 9871–5.
Johnstone, A. and Thorpe, R. (1982) Immunochemistry in Practice. Oxford: Blackwell Scientific Publications.
Keil, M., Sanchez-Serrano, J., Schell, J. and Willmitzer, L. (1986) Primary structure of a proteinase inhibitor II gene from potato (Solanum tuberosum).Nuc Acids Res. 14, 5641–50.
Kogan, M. (1986) Plant defense strategies and host-plant resistance. In Kogan, M. ed.,Ecological Theory and Integrated Post Management Practice, pp. 83–134. New York: John Wiley and Sons.
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature 727, 680–5.
MacIntosh, S.C., Kishore, G.M., Perlate, F.J., Marrone, P.G., Stone, T.B., Sims, S.R. and Fuchs, R.L. (1990) Potentiation ofBacillus thuringiensis insecticidal activity by serine protease inhibitors.J. Agric. Food Chem. 36, 1145–52.
Maniatis, T., Fritsch, E.F. and Sambrook, J. (1982)Molecular Cloning: a Laboratory manual. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press.
McManus, M.T., Tilson, V., White, D.W.R., Laing, W.A., Keen, G. and Christeller, J.T. (1990) Purification and characterisation of potato proteinase II from transgenic tobacco. InAbstracts from The New Zealand Genetical Society Meeting, Palmerston North, New Zealand. August 1990. A31.
McManus, M.T., Laing, W.A., Christeller, J.T. and White, D.W.R. (1993)Post-translational modification of an isoinhibitor from the potato proteinase inhibitor II gene family in transgenic tobacco yields a peptide with homology to potato chymotrypsin inhibitor I (PCI-1) (submitted).
Pearce, G., Sy, L., Russell, C., Ryan, C.A. and Hass, G.M. (1982) Isolation and characterisation from potato tubers of two polypeptide inhibitors of serine proteinases.Arch. Biochem. Biophys 213, 456–62.
Roberts, L.I.N. (1979) Biology ofChrysodeixis eriosoma (Lepidoptera: Noctuidae) in New Zealand.N. Z. Entomol. 7, 52–8.
Singh, P. (1983) A general purpose laboratory diet mixture for rearing insects.Insect Sci. Application 4, 357–62.
Spjotvoll, E. and Stoline, M.R. (1973) An extension of the T-method of multiple comparison to include the cases with unequal sample sizes.J. Am. Stat. Assoc. 68, 976–8.
Thornburg, R.W., An, G., Cleveland, T.E., Johnson, R. and Ryan, C.A. (1987) Wound-inducible expression of a potato proteinase inhibitor II-chloramphenicol acetyltransferase gene fusion in transgenic tobacco plants.Proc. Natl Acad. Sci. USA 84, 744–8.
Towbin, H., Stachelin, T. and Gordon, J. (1979) Electrophoretic transfer from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proc. Natl Acad. Sci. USA 76, 4350–4.
Vaeck, M., Reynaerts, A., Hofte, H., Jansens, S., Beucheleer, M. De, Dean, C., Zabeau, M., Montagu, M. van and Leemans, J. (1987) Transgenic plants protected from insect attack.Nature 328, 33–7.
Whiteley, J.R. and Schnepf, H.E. (1986) The molecular biology of parasporal crystal body formation inBacillus thuringiensis.Ann. Rev. Micriobiol. 40, 549–76.
Wolfson, J.L. and Murdock, L.L. (1987) Suppression of larval colorado potato beetle growth and development by digestive proteinase inhibitors.Entomol. Exp. Appl. 44, 235–40.