Comparative biochemistry of entomocidal parasporal crystals of selected Bacillus thuringiensis strains

Journal of Bacteriology - Tập 145 Số 2 - Trang 1052-1062 - 1981
D J Tyrell, Lee A. Bulla, Robert E. Andrews, Karl J. Kramer, Loren I. Davidson, P. Nordin

Tóm tắt

Parasporal crystals of Bacillus thuringiensis subspp. kurstaki, tolworthi, alesti, berliner, and israelensis were compared by electron microscopy, polyacrylamide gel electrophoresis, amino acid analysis, tryptic peptide mapping, immunological analysis, and insecticidal activity. Spore coats also were compared by polyacrylamide gel electrophoresis. B. thuringiensis subsp. israelensis crystals were lethally toxic to mosquito larvae and nontoxic to tobacco hornworm larvae. Conversely, crystals from the other subspecies killed tobacco hornworm larvae but were ineffective against mosquitoes. Crystalline inclusion bodies of all subspecies contained a protoxic subunit that had an apparent molecular weight of approximately 1.34 X 10(5). However, polyacrylamide gel electrophoretic patterns of solubilized crystals revealed a small-molecular-weight component (apparent molecular weight, 26,000) in B. thuringiensis subsp. israelensis that was absent in the other subspecies. Also, differences were noted in amino acid composition and tryptic peptide fingerprints. Crystal proteins were found in spore coats of all subspecies. The results suggest that insecticidal specificity is due to unique polypeptide toxins.

Từ khóa


Tài liệu tham khảo

Aronson , J. N. , and S. I. Fox . 1978 . Soluble glycoprotein insect toxin from the spore coat of Bacillus thuringiensis . Fed. Proc. 37 : 1824 .

Bechtel , D. B. , and L. A. Bulla , Jr. 1976 . Electron microscope study of sporulation and parasporal crystal formation in Bacillus thuringiensis . J. Bacteriol. 127 : 1472 - 1481 .

Bulla , L. A. , Jr. , G. A. Bennett , and 0. L. Shotwell . 1970 . Physiology of sporeforming bacteria associated with insects. II. Lipids of vegetative cells . J. Bacteriol. 104 : 1246 - 1253 .

Bulla , L. A. , Jr. , L. I. Davidson , K. J. Kramer , and B. L. Jones . 1979 . Purification of the insecticidal toxin from the parasporal crystal of Bacillus thuringiensis subsp. kurstaki . Biochem. Biophys. Res. Commun. 91 : 1123 - 1130 .

Bulla , L. A. , Jr. , K. J. Kramer , and L. I. Davidson . 1977 . Characterization of the entomocidal parasporal crystal of Bacillus thuringiensis . J. Bacteriol. 130 : 375 - 383 .

Cooksey K. E. 1971. The protein crystal toxin of Bacillus thuringiensis: biochemistry and mode of action p. 247-274. In H. D. Burges and N. W. Hussey (ed.) Microbial control of insects and mites. Academic Press Inc. New York.

Crestfield , A. M. , S. Moore , and W. H. Stein . 1963 . The preparation and enzymatic hydrolysis of reduced and Scarboxymethylated proteins . J. Biol. Chem. 238 : 622 - 627 .

de Barjac , H. 1978 . Un nouveau candidat a la Iutte biologique contre les moustiques: Bacillus thuringiensis var. israelensis . Entomophaga 23 : 309 - 319 .

de Barjac , H. 1978 . Une nouvelle variette de Bacillus thuringiensis tres toxique pour les Moustiques: B. thuringiensis var. israelensis serotype 14 . C. R. Acad. Sci. Ser. D 286 : 797 - 800 .

de Barjac , H. , and J. Coz . 1979 . Sensibilite comparees de six especes differentes de moustiques a Bacillus thuringiensisvar. israelensis . Bull. W. H. O. 57 : 139 - 141 .

Garcia , R. , and B. Desrochers . 1979 . Toxicity of Bacillus thuringiensis var. israelensis to some California mosquitoes under different conditions . Mosq. News 39 : 541 - 544 .

Goldberg , L. J. , and J. Margalit . 1977 . A bacterial spore demonstrating rapid larvicidal activity against Anopheles sergentii, Uranotaenia unguiculatata, Culex inivitatus, Aedes aegypti, and Culex pipiens . Mosq. News 37 : 355 - 358 .

Heilmann , J. , J. Barrollier , and E. Watzke . 1957 . Beitrag zur aminosaurebestimmung auf papierchromatogrammen . Hoppe Seyler's Z. Physiol. Chem. 309 : 219 - 220 .

Heinrikson , R. L. 1971 . The selective S-methylation of sulfhydryl groups in proteins and peptides with methylp-nitrobenzenesulfonate . J. Biol. Chem. 246 : 4090 - 4096 .

Hirs , C. M. W. 1967 . Determination of cystine as cysteic acid . Methods Enzymol. 11 : 59 - 62 .

Kramer , K. J. , R. L. Felsted , and J. H. Law . 1973 . Cocoonase. V. Structural studies on an insect serine protease . J. Biol. Chem. 248 : 3021 - 3028 .

Lecadet , M. M. , G. Chevrier , and R. Dedonder . 1972 . Analysis of a protein fraction in the spore coats of Bacillus thuringiensis . Eur. J. Biochem. 25 : 349 - 358 .

Liu , T. - Y. , and Y. H. Chang . 1971 . Hydrolysis of proteins with p-toluenesulfonic acid. Determination of tryptophan . J. Biol. Chem. 246 : 2842 - 2848 .

Lowry , 0. H. , N. J. Rosebrough , A. L. Farr , and R. J. Randall . 1951 . Protein measurement with the Folin phenol reagent . J. Biol. Chem. 193 : 265 - 275 .

Luft , J. 1961 . Improvements in epoxy resin embedding methods . J. Biophys. Biochem. Cytol. 9 : 409 - 414 .

Mopper , K. 1978 . Improved chromatographic separations on anion exchange resins . Anal. Biochem. 87 : 162 - 168 .

Nickerson , K. W. , G. St. Julian , and L. A. Bulla , Jr. 1974 . Physiology of sporeforming bacteria associated with insects: radiorespirometric survey of carbohydrate metabolism in the 12 serotypes of Bacillus thuringiensis . Appl. Microbiol. 28 : 129 - 132 .

Ohba , M. , and K. Aizawa . 1978 . Serological identification of Bacillu thuringiensis and related bacteria isolated in Japan . J. Invertebr. Pathol. 32 : 303 - 309 .

Ohba , M. and K. Aizawa . 1979 . A new subspecies of Bacillus thuringiensis possessing 11a:11c flagellar antigenic structure: Bacillus thuringiensis subsp. kyushuensis . J. Invertebr. Pathol. 33 : 387 - 388 .

Schesser , J. H. , and L. A. Bulla , Jr. 1978 . Toxicity of Bacillus thuringiensis spores to the tobacco hornworm Manduca sexta . Appl. Environ. Microbiol. 35 : 121 - 123 .

Schesser , J. H. , K. J. Kramer , and L. A. Bulla , Jr. 1977 . Bioassay for homogeneous parasporal crystal of Bacillus thuringiensis using the tobacco hornworm, Manduca sexta . Appl. Environ. Microbiol. 33 : 878 - 880 .

Sharpe , E. S. , K. W. Nickerson , L. A. Bulla , Jr. , and J. N. Aronson . 1975 . Separation of spores and parasporal crystals of Bacillus thuringiensis in gradients of certain X-ray contrasting agents . Appl. Microbiol. 30 : 1052 - 1053 .

Somerville , H. J. , F. P. Delafield , and S. C. Rittenberg . 1968 . Biochemical homology between crystal and spore protein of Bacillus thuringiensis . J. Bacteriol. 96 : 721 - 726 .

Takesaki , S. , and Kobata, A. 1974 . Microdetermination of individual neutral and amino sugars and N-acetylneuraminic acid in complex saccharides . J. Biochem. 76 : 783 - 789 .

Tyrell , D. J. , L. I. Davidson , L. A. Bulla , Jr. , and W. A. Ramoska . 1979 . Toxicity of parasporal crystals of Bacillus thuringiensis subsp. israelensis to mosquitoes . Appl. Environ. Microbiol. 38 : 656 - 658 .

Undeen , A. H. , and D. Berl . 1979 . Laboratory studies on the effectiveness of Bacillus thuringiensis var. israelensis de Barjac against Simulium damnosum (Diptera: Simuliidae) larvae . Mosq. News 39 : 742 - 745 .

Undeed , A. H. , and W. L Nagel . 1978 . The effect of Bacillus thuringiensis ONR-60A strain (Goldberg) on Simulium larvae in the laboratory . Mosq. News 38 : 524 - 527 .

Weber , K. , and M. Osborn . 1969 . The reliability of molecular weight determinations by dodecyl sulfatepolyacrylamide gel electrophoresis . J. Biol. Chem. 244 : 4406 - 4412 .

Weber , K. , J. R. Pringle , and M. Osborn . 1972 . Measurement of molecular weights by electrophoresis on SDS-acrylamide gel . Methods Enzymol. 26 : 3 - 27 .