Skip to main content
SpringerLink
Log in

A membrane technique for producing protoplasts ofCryphonectria parasitica

  • Published:
Current Microbiology Aims and scope Submit manuscript

Abstract

A method for the formation and regeneration of protoplasts of several strains of the chestnut blight fungus,Cryphonectria parasitica, is presented. The procedure utillizes cellophane membranes for growth and employs centrifugation for separation of protoplasts from hyphal fragments. Yields averaged 8.04×106 protoplasts per membrane. Regeneration frequencies were 40–50% with a soft-agar overlay. These protoplasts are suitable for use in experiments designed to determine the role of dsRNA in hypovirulence ofC. parasitica.

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

Literature Cited

  1. Anagnostakis SL (1984) Nuclear gene mutations inEndothia (Cryphonectria) parasitica that affect morphology and virulence. Phytopathology 74:561–565

    Google Scholar 

  2. Barrett V, Lemke PA, Dixon RK (1989) Protoplast formation from selected species of ectomycorrhizal fungi. Appl Microbiol Biotechnol 30:381–387

    Google Scholar 

  3. Dales RBG, Croft JH (1977) Protoplast fusion and the isolation of heterokaryons and diploids from vegetatively incompatible strains ofAspergillus nidulans. FEMS Microbiol Lett 1:201–204.

    Google Scholar 

  4. Day PR, Dodds JA, Elliston JE, Jaynes RA, Anagnostakis SL (1977) Double-stranded RNA inEndothia parasitica. Phytopathology 67:1393–1396

    Google Scholar 

  5. de Vries OMH, Wessels JGH (1972) Release of protoplasts fromSchizophyllum commune by a lytic enzyme preparation fromTrichoderma viride. J Gen Microbiol 73:13–22

    Google Scholar 

  6. Dodds JA (1980) Revised estimates of the molecular weights of dsRNA segments in hypovirulent strains ofEndothia parasitica. Phytopathology 70:1217–1220

    Google Scholar 

  7. Elliston JE (1985) Characteristics of dsRNA-free and dsRNA-containing strains ofEndothia parasitica in relation to hypovirulence. Phytopathology 75:151–158

    Google Scholar 

  8. Fincham JRS (1989) Transformation in fungi. Microbiol Rev 53:148–170 53:148–170

    Google Scholar 

  9. Gaugy D, Fevre M (1985) Regeneration and reversion of protoplasts from different species ofPenicillium. Microbios 44:285–293

    Google Scholar 

  10. Grente J, Berthelay-Sauret S (1978) Biological control of chestnut blight in France. In: MacDonald W, Chech F, Luchok J, Smith C (eds) Proceedings of the American Chestnut Symposium. W. Va Univ and US Forest Service, pp 30–34

  11. Hwang SC, Ko WH, Aragaki M (1975) A simplified method for sporangial production byPhytophthora cinnamomi. Mycologia 67:1233–1234

    Google Scholar 

  12. Jaynes RA, Elliston JE (1982) Hypovirulent isolates ofEndothia parasitica associated with large American chestnut trees. Plant Dis 66:769–772

    Google Scholar 

  13. Kitamoto Y, Mori N, Yamamoto M, Ohiwa T, Ichikawa Y (1988) A simple method for protoplast formation and improvement of protoplast regeneration from various fungi using an enzyme fromTrichoderma harzianum. Appl Microbiol Biotechnol 28:445–450

    Google Scholar 

  14. Kunkel W, Groth I, Jacob HE, Risch S, Harnisch M, May R, Berg H, Katenkamp U (1987) Electrofusion of protoplasts ofPenicillium chrysogenum. Stud Biophys 119:35–36

    Google Scholar 

  15. Layton AC, Kuhn DN (1988) Heterokaryon formation by protoplast fusion of drug-resistant mutants inPhytophthora megasperma f. sp.glycinea. Exp Mycol 12:180–194

    Google Scholar 

  16. Puhalla JE, Anagnostakis SL (1971) Genetics and nutritional requirements ofEndothia parasitica. Phytopathology 61:169–173

    Google Scholar 

  17. Sonnenberg ASM, Wessels JGH (1987) Heterokaryon formation in the basidiomyceteSchizophyllum commune by electrofusion of protoplasts. Theor Appl Genet 74:654–658

    Google Scholar 

  18. Van Alfen NK, Jaynes RA, Anagnostakis SL, Day PR (1975) Chestnut blight: biological control by transmissible hypovirulence inEndothia parasitica. Science 189:890–891

    Google Scholar 

  19. Yamada O, Magae Y, Kashiwagi Y, Kakimoto Y, Sasaki T (1983) Preparation and regeneration of mycelial protoplasts ofCollybia velutipes andPleurotus ostreatus. Eur J Appl Microbiol Biotechnol 17:298–300

    Google Scholar 

Download references

Author information

Author notes

  1. Jyan-Chyun Jang

    Present address: Department of Molecular Biology, Wellman 8th, Massachusetts General Hospital, 12114, Boston, MA, USA

Authors and Affiliations

  1. Department of Forest Resources, Clemson University, 29634-1003, Clemson, South Carolina, USA

    Jyan-Chyun Jang, Susan D. McElreath &  Frank H. Tainter

Authors
  1. Jyan-Chyun Jang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Susan D. McElreath
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Frank H. Tainter
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jang, JC., McElreath, S.D. & Tainter, F.H. A membrane technique for producing protoplasts ofCryphonectria parasitica . Current Microbiology 26, 113–116 (1993). https://doi.org/10.1007/BF01577346

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01577346

Keywords

Search

Navigation