Skip to main content

Fungal Protoplasts

  • Chapter
Genetics and Biotechnology

Part of the book series: The Mycota ((MYCOTA,volume 2))

Abstract

In the 40 years since the first reported isolation of protoplasts from filamentous or mycelial fungi (Peberdy 1971), these structures have become central to research in several key areas of fungal biology. The switch in focus in genetics from the classical to a molecular approach is the major factor. Two genetic techniques, protoplast fusion and transformation, together with electrophoretic techniques to determine the molecular karyotype of fungi are all dependent on the effective use of protoplasts. As a consequence, other aspects of research that employ protoplasts have benefited more generally. At the beginning, protoplasts held high curiosity value but their scientific potential was not fully realized. However, as the desire to extend the protoplast isolation technique to more and more fungi has occurred, correspondingly more attention has been given to improving this essential process. One outcome has been an overall improvement in protoplast yields for many fungi, which in turn has reawakened the interest of researchers addressing cell biological, physiological, and biochemical questions and led to the adoption of protoplasts as experimental tools wherever relevant.

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

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 74.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Andrade CM, Oliviera FM, Linardi VR (1992) Candida fennica: enhancement of protoplast formation and fusion. Can J Microbiol 8: 897–810

    Google Scholar 

  • Anné J, Peberdy JF (1985) Protopast fusion and inter-species hybridization in Penicillium. In: Peberdy JF, Ferenczy L (eds) Fungal protoplasts. Marcel Dekker, New York, pp 259–278

    Google Scholar 

  • Avram D, Petcu I, Radu M, Dan F, Stan R (1992) Electrically induced protoplast fusion for ergosterol-producing yeast strain improvement. J Basic Microbiol 32: 369–372

    Article  PubMed  CAS  Google Scholar 

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

    Article  Google Scholar 

  • Bennett JW (1985) Protoplasts and secondary metabolism. In: Peberdy JF, Ferenczy L (eds) Fungal protoplasts. Marcel Dekker, New York, pp 189–203

    Google Scholar 

  • Bigger CH, White MR, Braymer HD (1972) Ultrastructure and invertase secretion of the cot-2 mutant of Neurospora crassa. J Gen Microbiol 71: 159–166

    CAS  Google Scholar 

  • Bos CJ (1985) Protoplasts from fungal spores. In: Peberdy JF, Ferenczy L (eds) Fungal protoplasts. Marcel Dekker, New York, pp 73–86

    Google Scholar 

  • Bradshaw RE, Peberdy JF (1984) Protoplast fusion in Aspergillus: selection of interspecific heterokaryons using antifungal inhibitors. J Microbiol Methods 3: 27–32

    Article  Google Scholar 

  • Brigidi P, Matteuzzi D, Fava F (1988) Use of protoplast fusion to introduce methionine overproduction into Saccharomyces cerevisiae. Appl Microbiol Biotechnol 28: 268–271

    Article  CAS  Google Scholar 

  • Bulone V, Chanzy H, Gay L, Girard V, Fèvre M (1992) Characterization of chitin and chitin synthase from the cellulosic cell wall fungus Saprolegnia monoïca. Exp Mycol 16: 8–21

    Article  CAS  Google Scholar 

  • Chun SB, Chin JE, Bai S, An G-H (1992) Strain improvement of Phaffia rhodozyma by protoplast fusion. FEMS Microbiol Lett 93: 221–226

    Article  CAS  Google Scholar 

  • Daboussi MJ, Gerlinger G (1992) Parasexual cycle and genetic analysis following protoplast fusion in Nectria haematococca. Curr Genet 21: 385–392

    Article  CAS  Google Scholar 

  • Davis B (1985) Factors influencing protoplast isolation. In: Peberdy JF, Ferenczy L (eds) Fungal protopasts. Marcel Dekker, New York, pp 45–73

    Google Scholar 

  • Deed A, Seviour RJ, Swift IE (1990) Production of carotenoids by protoplasts of Acremonium diospyri. Enzyme Microb Technol 12: 940–944

    Article  CAS  Google Scholar 

  • Didek-Brumec M, Gaberc-Porekar V, Alacevié M, Sociè H (1993) Strain improvement of Claviceps Purpurea by protoplast fusion without introducing auxotrophic markers. Appl Microbiol Biotechnol 38: 746–749

    Article  CAS  Google Scholar 

  • Eggerding C, Randall JA, Sargent ML (1975) An altered invertase in the cot-2 mutant of Neurospora crassa. J Gen Microbiol 89: 102–112

    PubMed  CAS  Google Scholar 

  • Ferenczy L (1984) Fungal protoplast fusion: basic and applied aspects. In: Beers RE, Bassett EG (eds) Cell fusion: gene transfer and transformation. Raven Press, New York, pp 145–169

    Google Scholar 

  • Font de Mora J, Herrero E, Sentendreu R (1993) A kinetic study on the regeneration of Candida albicans protoplasts in the presence of cell wall synthesis inhibitors. FEMS Microbiol Lett 111: 43–48

    Google Scholar 

  • Fungaro MHP, de Azevedo JL, Pizzirani-Kleiner AA (1992) Genetic recombination after protoplast fusion in Candida sp. Rev Bras Genet 15: 499–507

    Google Scholar 

  • Furlaneto MC, Pizzirani-Kleiner AA (1992) Intraspecific hybridisation of Trichoderma pseudokoningii by anastomosis and protoplast fusion. FEMS Microbiol Lett 90: 191–196

    Article  Google Scholar 

  • Gabriel M, Kopeckâ M, Svoboda A (1992) Structural rearrangement of the actin cytoskeleton in regenerating protoplasts of budding yeasts. J Gen Microbiol 138: 2229–2234

    CAS  Google Scholar 

  • Gadau ME, Ling AJ (1992) Protoplast fusion in fungi. In: Arora DK, Elander RP, Mukerji KG (eds) Handbook of applied mycology. Vol 4: Fungal biotechnology. Marcel Dekker, New York, pp 101–128

    Google Scholar 

  • Gautam SP, Gupta AK, Pandey S (1993) Hybrid construction by fusion of protoplasts of Endomycopsis fibuligera and Saccharomyces cerevisiae. Cytobios 73: 183–188

    PubMed  CAS  Google Scholar 

  • Hamlyn PF, Bradshaw RE, Mellon FM, Santiago CM, Wilson JM, Peberdy JF (1981) Efficient protoplast isolation form fungi. Enzyme Microb Technol 3: 321–325

    Article  CAS  Google Scholar 

  • Hinnen A (1985) Some aspects of yeast transformation. In: Peberdy JF, Ferenczy L (eds) Fungal protoplasts. Marcel Dekker, New York, pp 323–336

    Google Scholar 

  • Hocart MJ, Peberdy JF (1990) Protoplasts and the improvement of fungi used in biological control. In: Whipps JM, Lumsden RD (eds) Biotechnology of fungi for improving plant growth. Cambridge University Press, Cambridge, pp 235–258

    Google Scholar 

  • Hoh YK, Tan TK, Yeoh H-H (1992) Protoplast fusion of ß-glucosidase producing Aspergillus niger strains. Appl Biochem Biotechnol 37: 81–88

    Article  PubMed  CAS  Google Scholar 

  • Jang J-C, McElreath SD, Tainter FH (1993) A membrane technique for producing protoplasts of Cryphonectria parasitica. Curr Microbiol 26: 113–116

    Article  CAS  Google Scholar 

  • Jochovâ J, Rupes I, Streiblovâ E (1991) F-actin contractile rings in protoplasts of the yeast Schizosaccharomyces. Cell Biol Int Rep 15: 607–610

    Article  PubMed  Google Scholar 

  • Jochovâ J, Rupes I, Peberdy JF (1993) The effect of the microtubule inhibitor benomyl on protein secretion in Aspergillus nidulans. Mycol Res 97: 23–27

    Article  Google Scholar 

  • Kaul W, Rossow U, Emeis C-C (1993) Screening for microorganisms with cell wall lytic activity to produce protoplast-forming enzymes. Appl Microbiol Biotechnol 39: 574–576

    Article  CAS  Google Scholar 

  • Kelkar HS, Shankar VS, Deshpande MV (1990) Rapid isolation and regeneration of Sclerotium rolfsii protoplasts and their potential application for starch hydrolysis. Enzyme Microb Technol 12: 510–514

    Article  CAS  Google Scholar 

  • Kevei F, Peberdy JF (1985) Interspecies hybridization after protoplast fusion in Aspergillus. In: Peberdy JF, Ferenczy L (eds) Fungal protoplasts. Marcel Dekker, New York, pp 241–258

    Google Scholar 

  • Kida K, Kume K, Morimuar S, Sonoda Y (1992) Repeated-batch fermentation process using a thermotolerant flocculating yeast constructed by protoplast fusion. J Ferment Bioeng 74: 169–173

    Article  CAS  Google Scholar 

  • Kinoshita Y, Hayase S, Higuchi M, Yamamoto Y, Ahmadjian V, Yoshimura I, Yamada Y (1991) Improvement of protoplast isolation from lichen mycobionts. Agric Biol Chem 55: 1891–1892

    Article  CAS  Google Scholar 

  • Kiramura K, Imura M, Lee SP, Kato Y, Usami S (1989) Intergeneric hybridization between Aspergillus niger and Trichoderma viride by protoplast fusion. Agric Biol Chem 53: 1589–1596

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Kolar H, Mischak H, Kammel WP, Kubicek CP (1985) Carboxymethylcellulase and ß-glucanase secretion by protoplasts of Trichoderma reesei. J Gen Microbiol 131: 1339–1347

    CAS  Google Scholar 

  • Kumari JA, Panda T (1992) Studies on critical analysis of factors influencing improved production of protoplasts from Trichoderma mycelium. Enzyme Microb Technol 14: 241–248

    Article  CAS  Google Scholar 

  • Kunkel W, Groth I, Jacob H-E, Risch S, Harnisch M, May R, Berg H, Katenkamp U (1987) Electrofusion of protoplasts of Penicillium chrysogenum. Stud Biophys 119: 1–3

    Google Scholar 

  • Lodder S, Wood D, Gull K (1993) A protoplasting technique with general applicability to molecular karyotyping of hymenomycetes. J Gen Microbiol 139: 1063–1067

    CAS  Google Scholar 

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

    Article  Google Scholar 

  • Lein J (1986) The Panlabs penicillin strain improvement programme. In: Vanek Z, Hostalek Z (eds) Overproduction of microbial metabolites: strain improvement and process control strategies. Butterworths, Boston, pp 105–139

    Google Scholar 

  • May G (1992) Fungal technology. In: Kinghorn JR, Turner G (eds) Applied molecular genetics of filamentous fungi. Blackie Academic & Professional, London, pp 1–27

    Chapter  Google Scholar 

  • Obraztsova IN, Nutsubidze NN, Prabakaran K, Demina VA, Gernet MV (1991) Production and fusion of pro 60 J.F. Peberdy: Fungal Protoplasts toplasts of fungus producers of lignolytic and cellulolytic enzymes, biochemical characteristics of the fusant enzyme complex. Microbiology (Eng! Trans!)

    Google Scholar 

  • Ogawa K, Brown JA, Wood TM (1987) Interspecific hybridization of Trichoderma reesei QM 9414 by protoplast fusion using colour mutants. Enzyme Microb Technol 9: 229–232

    Article  CAS  Google Scholar 

  • Peberdy JF (1971) Protoplasts from fungi. Sci Prog (Oxford) 60: 73–86

    Google Scholar 

  • Peberdy JF (1979a) Fungal protoplasts: isolation, reversion and fusion. Annu Rev Microbio! 33: 21–39

    Article  CAS  Google Scholar 

  • Peberdy JF (1979b) Wall biogenesis by protoplasts. In: Burnett JH, Trinci APJ (eds) Wall and hyphal growth. Cambridge University Press, Cambridge, pp 49–70

    Google Scholar 

  • Peberdy JF (1983) Genetic recombination in fungi by protoplast fusion and transformation. In: Smith JE (ed) Marcel Dekker, New York

    Google Scholar 

  • Peberdy JF (1985) Mycolytic enzymes. In: Peberdy JF, Ferenczy L (eds) Fungal protoplasts. Marcel Dekker, New York, pp 31–44

    Google Scholar 

  • Peberdy JF (1987a) Developments in protoplast fusion in fungi. Microbio! Sci 4: 108–111

    CAS  Google Scholar 

  • Peberdy JF (1987b) Genetics of Acremonium. In: Peberdy JF (ed) Penicillium and Acremonium: biotechnology handbooks. Plenum Press, New York, pp 93–111

    Google Scholar 

  • Peberdy JF (1989) Fungi without coats — protoplasts as tools for mycological research. Mycol Res 93: 1–20

    Article  Google Scholar 

  • Peberdy JF (1994) Protein secretion in fungi: trying to understand a highly productive black box. Trends Biotechnol 12: 50–57

    Article  PubMed  CAS  Google Scholar 

  • Peberdy JF, Ferenczy L (1985) Fungal protoplasts. Marcel Dekker, New York

    Google Scholar 

  • Peberdy JF, Hocart MJ (1987) Protoplasts as tools in the genetics of plant pathogenic fungi. In: Day PR, Jellis GJ (eds) Genetics and plant pathogenesis. Blackwell, Oxford, pp 127–141

    Google Scholar 

  • Ridout CJ, Coley-Smith JR, Lynch JM (1993) Fractionation of extracellular enzymes from a mycoparasitic strain of Trichoderma harzianum. Enzyme Microb Technol 10: 180–187

    Article  Google Scholar 

  • Roncal T, Ugalde UO, Barnes J, Pitt D (1991) Production of protoplasts of Penicillium cyclopium with improved viability and functional properties. J Gen Microbiol 137: 1647–1651

    CAS  Google Scholar 

  • Sanchez A, Larriba G, Villanueva JR, Villa TG (1980) Glycosylation is not necessary for the secretion of exo-1,3-ß-glucanase by Saccharomyces cerevisiae protoplasts. FEBS Lett 121: 283–286

    Article  CAS  Google Scholar 

  • Selitrennikoff CP, Bloomfield EC (1984) Formation and regeneration of protoplasts of wild-type Neurospora crassa. Curr Microbiol 11: 113–118

    Article  Google Scholar 

  • Sharma VK, Goel R, Johari BN (1988) Isolation, purification and regeneration of protoplasts from Sporotrichium thermophile conidiospores. Biochem Int 17: 895–906

    PubMed  CAS  Google Scholar 

  • Sienko M, Stepien P, Paszewski A (1992) Generation of genetic recombinants in Trichosporon cutaneum by spontaneous segregation of protoplast fusants. J Gen Microbiol 138: 1409–1412

    PubMed  CAS  Google Scholar 

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

    Article  Google Scholar 

  • Stovall ME (1992) Interstrain spheroplast fusion of Alternaria alternata. Appl Microbiol Biotechnol 37: 480–484

    Article  Google Scholar 

  • Tahoun MK (1993) Gene manipulation by protoplast fusion and penicillin production by Penicillium chrysogenum. Appl Biochem Biotechnol 39 /40: 445–453

    Article  PubMed  Google Scholar 

  • Tamova G, Betina V, Farkas V (1993) An efficient method for producing protoplasts from Trichoderma viride. Folia Microbiol 38: 214–218

    Article  CAS  Google Scholar 

  • Tashpulatov Zh, Shul’man TS, Baibaev BG, Mirzarakhimova M (1991) Formation and regeneration of protoplasts of the cellulolytically active fungus Trichoderma harzianum 19. Microbiology (Eng! Trans) 60: 377–380

    Google Scholar 

  • Torres KB, Bruckner B, Meier B (1992) Obtaining mutants for protoplast fusion of gibberellin-forming Gibberella fujikuroi strains. Appl Biochem Biotechnol 33: 83–95

    Article  PubMed  CAS  Google Scholar 

  • Toyama H, Toyama N (1990) Protoplast fusion between Aspergillus oryzae and Aspergillus niger in relation to their multinuclear nature. J Ferment Bioeng 69: 186–188

    Article  Google Scholar 

  • Toyama H, Yokoyama T, Shinmyo A, Okada H (1984) Interspecific protoplast fusion of Trichoderma. J Biotechnol 1: 25–35

    Article  Google Scholar 

  • Trevithick JR, Metzenberg RL (1966) Molecular sieving by Neurospora cell walls during secretion of invertase isoenzymes. J Bacteriol 92: 1010–1015

    PubMed  CAS  Google Scholar 

  • Tsukamoto C, Magae Y, Taniguchi H, Sasaki T (1990) Changes in the number of nuclei during formation of giant protoplasts in Pleurotus cornucopiae. J Gen Appl Microbio! 36: 451–454

    Article  Google Scholar 

  • Vagvölgyi Cs, Ferenczy L (1991) Isolation of nuclei from Aspergillus nidulans protoplasts. FEMS Microbio! Lett 82: 247–252

    Google Scholar 

  • Vagvölgyi Cs, Ferenczy L (1992) Transfer of isolated nuclei into protoplasts of Aspergillus nidulans. Experientia 48: 271–272

    Article  Google Scholar 

  • Vainstein MH, Peberdy JF (1991) Location of invertase in Aspergillus nidulans: release during hyphal wall digestion and secretion by protoplasts. Mycol Res 97: 1270–1274

    Article  Google Scholar 

  • van den Hondel CAMJJ, Punt PJ (1991) Gene transfer systems and vector development. In: Peberdy JF, Caten CE, Ogden JE, Bennett JW (eds) Applied molecular genetics of fungi. Cambridge University Press, Cambridge, pp 1–28

    Google Scholar 

  • Virk S, Johri BN, Singh SP (1992) Protoplasts from Malbranchea pulchella var. sulfurea: isolation and regeneration. J Gen Appl Microbiol 38: 75–78

    Article  CAS  Google Scholar 

  • Wangemann M, Schauz K (1989) Release of protoplasts from Ustilago maydis using Novozym 234. Angew Bot 63: 95–101

    Google Scholar 

  • Wellmann H, Shauz K (1993) DMI-resistance in Ustilago maydis. II. Effect of Triadimeton on regenerating protoplasts and analysis of fungicide uptake. Pestic Biochem Physiol 46: 55–64

    Google Scholar 

  • Yanagi SO, Monma M, Kawasumi T, Hino A, Kito M, Takebe I (1985) Conditions for isolation of and colony formation by mycelial protoplasts of Coprinus macrorhizus. Agric Biol Chem 49: 171–179

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1995 Springer-Verlag Berlin Heidelberg

About this chapter

Cite this chapter

Peberdy, J.F. (1995). Fungal Protoplasts. In: Kück, U. (eds) Genetics and Biotechnology. The Mycota, vol 2. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-662-10364-7_4

Download citation

  • DOI: https://doi.org/10.1007/978-3-662-10364-7_4

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-662-10366-1

  • Online ISBN: 978-3-662-10364-7

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics