Skip to main content
SpringerLink
Log in

Cuticular and non-cuticular substrate influence on expression of cuticle-degrading enzymes from conidia of an entomopathogenic fungus,Nomuraea rileyi

  • Human And Animal Mycology
  • Published:
Mycopathologia Aims and scope Submit manuscript

Abstract

Larval cuticle fromTrichoplusia ni, Helicoverpa (=Heliothis)zea, andHeliothis virescens and a cellulose substrate were used to quantify release of proteolytic, chitinolytic, and lipolytic enzymes by germinating conidia of the entomopathogenic fungus,Nomuraea rileyi. There was no significant difference in conidial viability incubated withT. ni, H. zea or cellulose substrates. Conidial viability onH. virescens cuticle, however, was significantly lower (ca. 19–25%) than the other three substrates. The presence of cuticle substrates, especially cuticle ofT. ni, stimulated germination. The nature of the substrate influenced both the time and quantity of the enzymes expressed. Specific proteases (aminopeptidase, chymoelastase, trypsin) generally were expressed earlier and/or in greater quantities on cuticular than on the cellulose substrate. Although both chitinolytic enzymes (endochitinase, N-acetylglucosaminidase) were detected on all three cuticular substrates, their activity was substantially lower than that of the proteolytic enzymes. Lipase activity was only minimally present. Early concurrent release of both proteases and chitinases suggested that both may be important in the penetration of the larval integument by germinating conidia ofN. rileyi. Expression of proteases and chitinases, especially aminopeptidase and endochitinase was probably a specific response to cuticle, because little or no activity was expressed on the non-host, cellulose substrate.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Steinhaus EA, Marsh GA. Report of diagnosis of diseased insects 1951–1961. Hilgardia 1962; 33: 349–490.

    Google Scholar 

  2. Gabriel BP: Enzymatic activities of some entomophthorous fungi. J Invertebr Pathol 1968; 11: 70–81.

    Google Scholar 

  3. Charnley AK. Physiological aspects of destructive pathogenesis in insects by fungi: A speculative review. In Anderson JM, Rayner ADM, Walton DWH, (eds.). Invertebrate-Microbial Interactions. Brit Mycol Soc Symp 6. London: Cambridge Univ Press, 1984; 229–70.

    Google Scholar 

  4. Hüber J. Untersuchugen zur Physiologie insektentotender Pilze. Arch Mikrobiol 1958; 29: 257–76.

    Google Scholar 

  5. Gabriel BP. Histochemical study of the insect cuticle infected by the fungusEntomophthora coronata. J Invertebr Pathol 1968; 11: 82–89.

    Google Scholar 

  6. Leopold J, Samsinakova A. Quantitative estimation of chitinase and several other enzymes in the fungusBeauveria bassiana. J Invertebr Pathol 1970; 15: 34–42.

    Google Scholar 

  7. Mohamed AKA, Turner AG. Proteolytic activity ofNomuraea rileyi on casein and host insect cuticle. Mycopathologia 1983; 13–15.

  8. St. Leger RJ, Cooper RM, Charnley AK. Production of cuticle-degrading enzymes by the entomopathogenMetarhizium ansiopliae during infection of cuticle fromCalliphora vomitaria andManduca sexta. J Gen Microbiol 1987; 133: 1371–82.

    Google Scholar 

  9. El-Sayed GN, Ignoffo CM, Leathers TD, Gupta SC. Use of colorometric system to detect enzymes expressed by germinating conidia of entomopathogenic fungi. Mycopathologia 1992; 118: 29–36.

    Google Scholar 

  10. Soderhall K, Svensson E, Unestam T. Chitinase and protease activity in germinating zoospore cyst of a parasitic fungus,Aphanomyces astaci, Oomycetes. Mycopathologia 1978; 64: 9–11.

    Google Scholar 

  11. Balasubramanian R, Manocha MS. Proteinase, chitinase and chitosanase activities in germinating spores ofPiptocephalis virginiana. Mycologia 1986; 78: 157–63.

    Google Scholar 

  12. Goettel MS, St. Leger RJ, Rizzo NW, Staples RC, Roberts DW. Ultrastructural localization of a cuticle-degrading protease produced by the entomopathogenic fungusMetarhizium anisopliae during penetration of host (Manduca sexta) cuticle. J Gen Microbiol 1989; 135: 2233–39.

    Google Scholar 

  13. St. Leger RJ, Goettel M, Roberts DW, and Staples RC. Prepenetration events during infection of host cuticle byMetarhizium anisopliae. J Invertebr Pathol 1991; 58: 168–79.

    Google Scholar 

  14. Ignoffo CM, Garcia C. Host spectrum and relative virulence of an Ecuadoran and Mississippian biotype ofNomuraea rileyi. J Invertebr Pathol 1985; 45: 346–52.

    Google Scholar 

  15. Ignoffo CM, Boucias DB. Relative activity of geographical isolates ofNomuraea bioassayed against the cabbage looper and velvet bean caterpillar. J Invertebr Pathol 1992; 59: 215–17.

    Google Scholar 

  16. Ignoffo CM. Insect viruses. In Smith CN, ed, Insect Colonization and Mass Production. New York: Academic Press, 1966: 501–30.

    Google Scholar 

  17. Hackman RH. Biochemical methods (proteins). In Miller TA, ed. Cuticle Techniques in Arthropods. New York: Springer Verlag, 1980: 145–184.

    Google Scholar 

  18. Mishell BB, Shiigi SM. Selected Methods in Cellular Immunology. San Francisco: Freeman, 1980: 16.

    Google Scholar 

  19. Bradford M. A rapid and sensitive method for the quantification of microgram quantities of proteins utilizing the principle of protein-dye-binding. Anal Biochem 1976; 72: 248–54.

    Google Scholar 

  20. Bidachoka MJ, Khahatourians GG. Identification ofBeauveria bassiana extracellular protease as a viruelence factor in pathogenicity toward the migratory grasshopper,Melanoplus sanguinipes. J Invertebr Pathol 1990; 56: 362–370.

    Google Scholar 

  21. Frugoni JAC. Tampone universal di britton e robinson a forza ionica constante. Gazz Chim Ital 1957; 87: 403–405.

    Google Scholar 

  22. Erlanger BF, Kokowski N, Cohen W. The preparation and properties of two new chromogenic substrates of trypsin. Arch Biochem Biophys 1961, 95; 271–78.

    Google Scholar 

  23. Nakadai T, Nasuno S, Iguchi. N. Purification and properties of leucine aminopeptidase I fromAspergillus oryzae. Agr Biol Chem 1973; 37: 757–765.

    Google Scholar 

  24. El-Sayed GN, Coudron TA, Ignoffo CM, Riba G. Chitinolytic activity and virulence associated with native and mutant isolates of an entomopathogenic fungus,Nomuraea rileyi (Farlow) Samson. J Invertebr Pathol 54: 394–403.

  25. Bender ML, Kedzy FJ, Wedler FC. α-Chymotrypsin: Enzyme concentration and kinetics. J Chem Educ 1967; 44: 84–88.

    Google Scholar 

  26. Tetz NW. Measurement of lipase activity in serum. In Cooper GR, ed. Standard Methods of Clinical Chemistry, Vol. 7. New York: Academic Press, 1972: pp. 19–31.

    Google Scholar 

  27. Smith RJ, Grula EA. Nutritional requirements for conidial germination and hyphal growth ofBeauveria bassiana. J. Invertebr Pathol 37: 222–230.

  28. Coudron TA, Kroha MJ, Ignoffo CM. Levels of chitinolytic activity during development of three entomopathogenic fungi. Comp Biochem Physiol B 1984; 79: 339–48.

    Google Scholar 

  29. Kerwin JL. Fatty acid regulation of the germination ofErynia variabilis conidia on adults and puparia of the lesser housefly,Fannia canicularis. Can J Microbiol 1984; 30: 158–61.

    Google Scholar 

  30. Latgé JP, Samperdo L, Brey P, Diaquin M. Agressiveness ofConidiobolus obscurus against the pea aphid: Influence of cuticular extracts on ballistopore germination of aggressive and non-aggressive strains. J Gen Microbiol 1987; 133: 1987–1997.

    Google Scholar 

  31. El-Sayed GN, Ignoffo CM, Leathers TD. Effects of cuticle source and concentration on germination of conidia of two isolates ofNomuraea rileyi. Mycopathologia 1991; 113: 95–102.

    Google Scholar 

  32. Hoffman-Ostenhof O. Enzymes inhibited by quinones. In Hochster RM, Quastal JH, eds. Metabolic Inhibitors. New York: Academic Press, 1963: 145–59.

    Google Scholar 

  33. Kuo MJ, Alexander M. Inhibition of lysis of fungi by melanin. J Bacteriol 1967; 94: 624–629.

    Google Scholar 

  34. Bull AT. Inhibition of polysaccharases by melanin: Enzyme inhibition in relation to mycolysis. Arch Biochem Biophys 1970; 137: 345–56.

    Google Scholar 

  35. Hackman RH. The integument of Arthropoda. In Florkin M, Scheer BT, eds. Chemical Zoology. New York: Academic Press, 1971: 1–53.

    Google Scholar 

  36. Michalski C, Domnas AJ. The beta D glucosidase system of an Actinoplanes sp. Carbohydr. Res. 1974; 33: 153–170.

    Google Scholar 

  37. Kucera M. Inhibition of the toxic proteases fromMetarhizium ansiopliae by extracts ofGalleria mellonella larvae. J Invertebr Pathol 1982; 40: 299–300.

    Google Scholar 

  38. Bell TJ, Lee B, Domnas AJ. The extracellular acid phosphatase of the mosquito-parasitizing fungusLagenidium giganteum. J Invertebr Pathol 1989; 54: 306–313.

    Google Scholar 

  39. Bidochka MJ, Khachatourians GG. N-acetyl-D-glucosa-mine-mediated regulation of extracellular protease in the entomopathogenic fungusBeauveria bassiana. Appl Environ Microbiol 1988; 54: 2699–2704.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biological Control of Insects Research Laboratory, USDA/ARS, Research Park, Rt. K, P.O. Box 7629, 65205, Colombia, MO, USA

    G. N. El-Sayed & C. M. Ignoffo

  2. Microbial Properties Research Center, USDA/ARS, Peoria, Ill, USA

    T. D. Leathers & S. C. Gupta

Authors
  1. G. N. El-Sayed
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C. M. Ignoffo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. T. D. Leathers
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S. C. Gupta
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This article reports the results of research only. Mention of a proprietary product in this paper does not constitute a recommendation for use by the US Department of Agriculture.

Rights and permissions

Reprints and permissions

About this article

Cite this article

El-Sayed, G.N., Ignoffo, C.M., Leathers, T.D. et al. Cuticular and non-cuticular substrate influence on expression of cuticle-degrading enzymes from conidia of an entomopathogenic fungus,Nomuraea rileyi . Mycopathologia 122, 79–87 (1993). https://doi.org/10.1007/BF01103603

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01103603

Key words

Search

Navigation