1887

Abstract

subsp. IPS78 and . HD133 both secreted exochitinase activity when grown in a medium containing chitin. Allosamidin, a specific chitinase inhibitor, inhibited activity from both strains, with IC values of about 50 μM with colloidal chitin as substrate and between 1 and 10 μM with 4-methylumbelliferyl-diacetylchitobioside and 4-methylumbelliferyl-triacetylchitotrioside as substrates. The involvement of these chitinolytic activities during pathogenesis in insects has been investigated with subsp. IPS78 against larvae of the midge , and with subsp. HD133 against caterpillars of the cotton leafworm . Presence of 100 μM allosamidin increased the LD by factors of 1.3 and 1.4, respectively, demonstrating a role for bacterial chitinases in the attack on the insects. Presence of chitinase A from considerably decreased the values for LD' confirming previous observations with different systems of the potentiation of entomopathogenesis of by exogenous chitinases. The most likely action of the endogenous chitinases of is to weaken the insects' peritrophic membranes, allowing more ready access of the bacterial toxins to the gut epithelia. Addition of exogenous chitinases will then increase this effect. Complementary cross-infection experiments, strain HD133 against midge larvae and strain IPS78 against caterpillars, were performed to investigate the pathogen/host specificities of the effects. Results showed that much higher concentrations of bacteria were required to achieve even low mortalities, and addition of chitinase A gave no increase in death rate.

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1998-08-01
2024-03-28
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References

  1. Alam M., Mizutani T., Isono M., Nikaidou N., Watanabe T. 1996; Three chitinase genes (chiA, chiB and chiC) comprise the chitinase system of Bacillus circulans WL-12. . J Ferment Bioeng 82:28–36
    [Google Scholar]
  2. Bot J. 1967; Artificial rearing method for three noctuids of economical importance belonging to the genus Spodoptera. . J Entomol Soc Sth Afr 29:157–160
    [Google Scholar]
  3. Carter D.J. 1984; Spodoptera littoralis. In Pest Lepidoptera of Europe, . pp. 274–275 Carter D. J. Edited by Boston: Dr W. Junk Publishers;
    [Google Scholar]
  4. deBarjac H. 1981; Identification of H-serotypes of Bacillus thuringiensis. In Microbial Control of Pests and Plant Diseases, 1970-1980 . pp. 35–43 Burges H. W. Edited by London: Academic Press;
    [Google Scholar]
  5. deBarjac H. 1990; Characterisation and prospective view of Bacillus thuringiensis israelensis. In Bacterial Control of Mosquito and Blackflies, . pp. 10–15 deBarjac H., Sutherland D. J. Edited by Rutgers: Rutgers University Press;
    [Google Scholar]
  6. Finney D.J. 1962; Probit Analysis,3rd edn. Cambridge: Cambridge University Press..
    [Google Scholar]
  7. Fuhrman J.A., Lee J., Dalamagas D. 1995; Structure and function of a family of chitinase isozymes from Brugian micro-filariae. Exp Parasitol . 80:672–680
    [Google Scholar]
  8. Gooday G.W. 1990; Inhibition of chitin metabolism. In The Biochemistry of Cell Walls and Membranes in Fungi, . pp. 61–79 Kuhn P. J., Trinci A. P. J., Jung M. J., Trinci M. W., Goosey A. P. J., Copping L. G. Edited by Berlin: Springer;
    [Google Scholar]
  9. Gooday G.W. 1994; Physiology of microbial degradation of chitin and chitosan. In Biochemistry of Microbial Degradation, . pp. 279–312 Ratledge C., Bhattacharya SK. Edited by Dordrecht: Kluwer;
    [Google Scholar]
  10. Gooday G.W., Brydon L.J., Chappell L.H. 1988; Chitinase in female Onchocerca gibsoni and its inhibition by allosamidin.. Mol Biochem Parasitol 29:223–225
    [Google Scholar]
  11. Hawtin R.E., Zarkowska T., Arnold K., Thomas C.A., Gooday G.W., King L.A., Kuzio J.A. 1997; Liquefaction of Autographa californica nucleopolyhedrovirus-infected insects is dependent on the integrity of virus-encoded chitinase and cathepsin genes.. Virology 238:243–253
    [Google Scholar]
  12. Huber M., Cabib E., Miller L. 1991; Malaria parasite chitinase and penetration of the mosquito peritrophic membrane. Proc Natl Acad Sci USA . 88:2807–2810
    [Google Scholar]
  13. Kettle D.S. 1977; Biology and bionomics of bloodsucking ceratopogonids. . Annu Rev Entomol 22:33–51
    [Google Scholar]
  14. Kuranda M.J., Robbins P.W. 1987; Cloning and heterologous expression of glycosidase genes from Saccbaromyces cerevisiae. . Proc Natl Acad Sci USA 84:2585–2589
    [Google Scholar]
  15. McCreath K.J., Gooday G.W. 1992; A rapid and sensitive micro-assay for determination of chitinolytic activity.. J Microbiol Methods 14:229–237
    [Google Scholar]
  16. Milewski S., O'Donnell R., Gooday G.W. 1992; Chemical modification studies of the active centre of Candida albicans chitinase and its inhibition by allosamidin.. J Gen Microbiol 138:2545–2550
    [Google Scholar]
  17. Molano J., Duran A., Cabib E. 1977; A rapid and sensitive assay for chitinase using tritiated chitin.. Anal Biochem 83:648–656
    [Google Scholar]
  18. Regev A., Keller M., Strizhov N., Sneh B., Prudovsky E., Chet I., Ginzberg I., Koncz-Kalman Z., Koncz C., Schell J., Zilberstein A. 1996; Synergistic activity of a Bacillus thuring- iensis△-endotoxin and a bacterial endochitinase against Spodop- tera littoris larvae. . Appl Environ Microbiol 62:3581–3586
    [Google Scholar]
  19. Sakuda S., Isogai A., Matsumoto S., Suzuki A. 1987a; Search for microbial insect growth regulators. II. Allosamidin, a novel insect chitinase inhibitor.. J Antibiot 62:40296–300
    [Google Scholar]
  20. Sakuda S., Isogai A., Matsumoto S., Makita T., Matsumoto S., Koseki K., Kodama H., Suzuki A. 1987b; Structures of allosamidins, novel insect chitinase inhibitors, produced by actinomycetes.. Agric Biol Chem 51:3251–3259
    [Google Scholar]
  21. Sampson M.N., Gooday G.W. 1996; A novel chitinase assay using the fluorescent brightener Calcofluor White M2R. In Chitin Enzymology, . 2: pp. 227–234 Muzzarelli R. A. A. Edited by Grottammare: Atec Edizioni;
    [Google Scholar]
  22. Schlein Y., Raymond L.J., Shlomai J. 1991; Chitinase secreted by Leishmania functions in sandfly vector.. Proc R Soc Lond Ser B 245:121–126
    [Google Scholar]
  23. Shahabuddin M., Kaslow D.C. 1993; Chitinase: a novel target for blocking parasite transmission? . Parasitol Today 9:252–255
    [Google Scholar]
  24. Shahabuddin M., Toyoshima T., Aikawa M., Kaslow D.C. 1993; Transmission-blocking activity of a chitinase inhibitor and activation of malarial parasite chitinase by mosquito protease. . Proc Natl Acad Sci USA 90:4266–4270
    [Google Scholar]
  25. Shapiro M., Preisler H.K., amp;Robertson J.L. 1987; Enhancement of baculovirus activity on gypsy moth (Lepidoptera: Lyman- triidae) by chitinase.. J Econ Entomol 80:1113–1116
    [Google Scholar]
  26. Smirnoff W.A. 1974; Three years of aerial field experiments with Bacillus thuringiensis plus chitinase formulation against the spruce budworm.. J Invertebr Pathol 24:344–348
    [Google Scholar]
  27. Sneh B., Schuster S., Gross S. 1983; Improvement of the insecticidal activity of Bacillus thuringiensis var. entomocidus on larvae of Spodoptera littoralis (Lepidoptera, Noctuidae) by addition of chitinolytic bacteria, a phagostimulant and a UV- protectant.. Z Angew Entomol 96:77–83
    [Google Scholar]
  28. Somers P.J.B., Yao R.C., Doolin L.R., McGowan M.J., Fakuda D.S., Mynderse I.S. 1987; Method for the detection and quantification of chitinase inhibitors in fermentation broths; isolation and insect life cycle. Effect of A82516.. J Antibiot 40:1751–1756
    [Google Scholar]
  29. Trudel J., Asselin A. 1989; Detection of chitinase activity after polyacrylamide gel electrophoresis. . Anal Biochem 178:362–366
    [Google Scholar]
  30. Welburn S.G, Maudlin I., amp;Gooday G.W. 1993; Rickettsia-like organisms and chitinase production in relation to transmission of trypanosomes by tsetse flies.. Parasitology 107:141–145
    [Google Scholar]
  31. Wiwat G, Siwayapram P., Pantu-watana S., Bhumiratana A. 1996; Expression of chitinase- encoding genes from Aeromonas hydrophila and Pseudomonas maltophila in Bacillus thuringiensis subsp. israelensis. . Gene 179:119–126
    [Google Scholar]
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