Skip to main content
SpringerLink
Log in

Isolation of Mutants and Construction of Intersexual Heterokaryons of Blakeslea trispora

  • Protocol
  • First Online:
Microbial Carotenoids From Fungi

Part of the book series: Methods in Molecular Biology ((MIMB,volume 898))

Abstract

The Mucoral fungus Blakeslea trispora is used for the industrial production of β-carotene and lycopene. Two genetic techniques have been used to increase carotene accumulation: the isolation of mutants and the formation and segregation of heterokaryons. Because all life stages are multinucleated, recessive mutants are isolated after exposure to N-methyl-N′-nitro-N-nitrosoguanidine, a strong mutagen and inactivator of nuclei. Intersexual heterokaryons are obtained easily, because they are formed spontaneously during sexual interaction. Here are the pertaining methods, based on those previously developed for Phycomyces blakesleeanus, a related and better-known fungus.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Ciegler A (1965) Microbial carotenogenesis. Adv Appl Microbiol 7:1–34

    Article  PubMed  CAS  Google Scholar 

  2. Cerdá-Olmedo E, Lipson ED (eds) (1987) Phycomyces. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  3. Cerdá-Olmedo E (2001) Phycomyces and the biology of light and color. FEMS Microbiol Rev 25:503–512

    Article  PubMed  Google Scholar 

  4. Mehta BJ, Obraztsova IN, Cerdá-Olmedo E (2003) Mutants and intersexual heterokaryons of Blakeslea trispora for production of β-carotene and lycopene. Appl Environ Microbiol 69:4043–4048

    Article  PubMed  CAS  Google Scholar 

  5. Avalos J, Cerdá-Olmedo E (2004) Fungal carotenoid production. In: Arora DK (ed) Handbook of fungal biotechnology, vol 20. Marcel Dekker, New York, pp 367–378

    Google Scholar 

  6. Rodríguez-Sáiz M, Paz B, De la Fuente JL, López-Nieto MJ, Cabri W, Barredo JL (2004) Blakeslea trispora genes for carotene biosynthesis. Appl Environ Microbiol 70:5589–5594

    Article  PubMed  Google Scholar 

  7. Silva F, Navarro E, Peñaranda A, Murcia-Flores L, Torres-Martínez S, Garre V (2008) A RING-finger protein regulates carotenogenesis via proteolysis-independent ubiquitylation of a white collar-1-like activator. Mol Microbiol 70:1026–1036

    PubMed  CAS  Google Scholar 

  8. Cerdá-Olmedo E (1975) The genetics of Phycomyces blakesleeanus. Genet Res 25:285–296

    Article  PubMed  Google Scholar 

  9. Silva F, Torres-Martínez S, Garre V (2006) Distinct white collar-1 genes control specific light responses in Mucor circinelloides. Mol Microbiol 61:1023–1037

    Article  PubMed  CAS  Google Scholar 

  10. Haro JP, Calo S, Cervantes M, Nicolás FE, Torres-Martínez S, Ruiz-Vázquez RM (2009) A single dicer gene is required for efficient gene silencing associated with two classes of small antisense RNAs in Mucor circinelloides. Eukaryot Cell 8:1486–1497

    Article  PubMed  Google Scholar 

  11. Mehta BJ, Cerdá-Olmedo E (1995) Mutants of carotene production in Blakeslea trispora. Appl Microbiol Biotechnol 42:836–838

    Article  CAS  Google Scholar 

  12. Cerdá-Olmedo E, Reau P (1970) Genetic classification of the lethal effects of various agents on heterokaryotic spores of Phycomyces. Mutat Res 9:369–384

    Article  PubMed  Google Scholar 

  13. Roncero MIG, Zabala C, Cerdá-Olmedo E (1984) Mutagenesis in multinucleate cells: the effects of N-methyl-N′-nitro-N-nitrosoguanidine on Phycomyces spores. Mutat Res 125:195–204

    Article  PubMed  CAS  Google Scholar 

  14. De la Guardia MD, Aragón CMG, Murillo FJ, Cerdá-Olmedo E (1971) A carotenogenic enzyme aggregate in Phycomyces: evidence from quantitative complementation. Proc Natl Acad Sci USA 68:2012–2015

    Article  PubMed  Google Scholar 

  15. Burgeff H (1924) Untersuchungen über Sexualität und Parasitismus bei Mucorineen 1. Botanische Abhandlungen 4:1–135

    Google Scholar 

  16. Polaino S, Herrador MM, Cerdá-Olmedo E, Barrero AF (2010) Splitting of β-carotene in the sexual interaction of Phycomyces. Org Biomol Chem 8:4229–4231

    Article  PubMed  CAS  Google Scholar 

  17. Sutter RP (1970) Trisporic acid synthesis in Blakeslea trispora. Science 168:1590–1592

    Article  PubMed  CAS  Google Scholar 

  18. Kuzina V, Cerdá-Olmedo E (2006) Modification of sexual development and carotene production by acetate and other small carboxylic acids in Blakeslea trispora and Phycomyces blakesleeanus. Appl Environ Microbiol 72:4917–4922

    Article  PubMed  CAS  Google Scholar 

  19. Burgeff H (1914) Untersuchungen über Variabilität, Sexualität und Erblichkeit bei Phycomyces nitens Kunze, I. Flora 107:259–316

    Google Scholar 

  20. Murillo FJ, Calderón IL, López-Díaz I, Cerdá-Olmedo E (1978) Carotene-superproducing strains of Phycomyces. Appl Environ Microbiol 36:639–642

    PubMed  CAS  Google Scholar 

  21. Gauger WL, Peláez MI, Álvarez MI, Eslava AP (1980) Mating type heterokaryons in Phycomyces blakesleeanus. Exp Mycol 4:56–64

    Article  Google Scholar 

  22. Thaxter R (1914) New or peculiar zygomycetes. III. Blakeslea, Dissophora, Haplospo­rangium, nova genera. Bot Gaz (Crawfordsville) 58:353–366

    Article  Google Scholar 

  23. Zycha H, Siepmann R, Linnemann G (1969) Mucorales, Eine Beschreibung aller Gattungen und Arten dieser Pilzegruppe. Cramer, Lehre

    Google Scholar 

Download references

Acknowledgment

This work was supported by Junta de Andalucía (CVI-03901) and the Spanish Government (BIO2009-12486 and 11131).

Author information

Authors and Affiliations

  1. Departamento de Genética, Facultad de Biología, Universidad de Sevilla, Sevilla, Spain

    Enrique Cerdá-Olmedo & Bina J. Mehta

Authors
  1. Enrique Cerdá-Olmedo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bina J. Mehta
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Enrique Cerdá-Olmedo .

Editor information

Editors and Affiliations

  1. Parque Tecnológico de León, Gadea Biopharma, Boecillo 47151, Valladolid, 24009, León, Spain

    José-Luis Barredo

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media New York

About this protocol

Cite this protocol

Cerdá-Olmedo, E., Mehta, B.J. (2012). Isolation of Mutants and Construction of Intersexual Heterokaryons of Blakeslea trispora . In: Barredo, JL. (eds) Microbial Carotenoids From Fungi. Methods in Molecular Biology, vol 898. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-918-1_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-918-1_4

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-917-4

  • Online ISBN: 978-1-61779-918-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation