, Volume 70, Issue 3, pp 306–310 | Cite as

Screening for Ergot Alkaloid Producers among Microscopic Fungi by Means of the Polymerase Chain Reaction

  • L. V. Boichenko
  • D. M. Boichenko
  • N. G. Vinokurova
  • T. A. Reshetilova
  • M. U. Arinbasarov


The potential of the polymerase chain reaction for the detection of ergot alkaloid producers among microscopic fungi of the generaPenicilliumand Clavicepswas evaluated. Twenty-three strains of various species of fungi with a previously studied capacity for alkaloid production were used. The internal fragment of the gene encoding 4-dimethylallyltryptophan synthase, the enzyme catalyzing the first step in the biosynthesis of ergot alkaloids, was amplified using degenerate primers. This approach revealed an about 1.2-kb specific DNA fragment in micromycetes synthesizing ergot alkaloids with complete tetracyclic ergoline system. Microorganisms that produce alkaloids with modified C or D ergoline rings, as well as α-cyclopiazonic acid, did not yield the PCR fragment of the expected size. This fragment was also not found in fungi incapable of ergot alkaloid production.

alkaloids microscopic fungi PCR screening 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Fliger, M., Wirst, M., and Shelby, R., Ergot Alkaloids: Sources, Structures and Analytical Methods, Folia Microbiol., 1997, vol. 42, no. 1, pp. 3–30.Google Scholar
  2. 2.
    Rehacek, Z. and Sajdl, P., Ergot Alkaloids, Prague: Academia, 1990.Google Scholar
  3. 3.
    Shapira, R., Paster, N., Eyal, O., Menasherov, M., Mett, A., and Salomon, R., Detection of Aflatoxigenic Molds in Grains by PCR, Appl. Environ. Microbiol., 1996, vol. 62, no. 9, pp. 3270–3273.Google Scholar
  4. 4.
    Cenis, J.L., Rapid Extraction of Fungal DNA for PCR Amplification, Nucleic Acids Res., 1992, vol. 20, p. 2380.Google Scholar
  5. 5.
    Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor: Cold Spring Harbor Lab. Press, 1989.Google Scholar
  6. 6.
    Kozlovskii, A.G., Arinbasarov, M.U., Solov'eva, T.F., Adanin, V.M., Grigorov, I., Angelov, T.I., Slokoska, L.S., and Angelova, M.B., Alkaloids of Claviceps sp. IBPM F-401, Prikl. Biokhim. Mikrobiol., 1980, vol. 16, no. 4, pp. 569–577.Google Scholar
  7. 7.
    Tudzynski, P., Holter, K., Correia, T., Arntz, C., Grammel, N., and Keller, U., Evidence for an Ergot Alkaloid Gene Cluster in Claviceps purpurea, Mol. Gen. Genet., 1999, vol. 261, no. 1, pp. 133–141.Google Scholar
  8. 8.
    Solov'eva, T.F., Baskunov, B.P., Kiseleva, O.V., and Kozlovskii, A.G., Biosynthesis of Alkaloids by Penicillium puberulum, Mikrobiologiya, 1992, vol. 61, no. 3, pp. 395–403.Google Scholar
  9. 9.
    Kozlovskii, A.G., Vinokurova, N.G., Zhelifonova, V.P., and Ozerskaya, S.M., Alkaloid Formation by Penicillia of the Series Fellutana and Canescentia, Mikrobiologiya, 1997, vol. 66, no. 4, pp. 514–519.Google Scholar
  10. 10.
    Kozlovskii, A.G., Vinokurova, N.G., and Ozerskaya, S.M., Peculiarities of Alkaloid Formation in Penicillium chrysogenum Strains Isolated from Soils of Different Climatic Zones, Mikrobiologiya, 1998, vol. 67, no. 4, pp. 483–487.Google Scholar
  11. 11.
    Kozlovskii, A.G., Stefanova-Avramova, L.N., Reshetilova, T.A., Sakharovskii, V.G., and Adanin, V.M., Clavine Ergot Alkaloids Produced by Penicillium gorlencoanum, Prikl. Biokhim. Mikrobiol., 1981, vol. 17, no. 6, pp. 806–812.Google Scholar
  12. 12.
    Vinokurova, N.G., Reshetilova, T.A., Adanin, V.M., and Kozlovskii, A.G., Alkaloid Composition of the Fungi Penicillium palitans and Penicillium oxalicum, Prikl. Biokhim. Mikrobiol., 1991, vol. 27, no. 6, pp. 850–855.Google Scholar
  13. 13.
    Kozlovskii, A.G., Reshetilova, T.A., Sakharovskii, V.G., Adanin, V.M., and Zyakun, A.M., Products of the Metabolism of the Alkaloids Roquefortine and 3,12-Dihydroroquefortine by the Fungus Penicillium farinosum, Prikl. Biokhim. Mikrobiol., 1988, vol. 24, no. 5, pp. 642–646.Google Scholar
  14. 14.
    Kozlovskii, A.G., Vinokurova, N.G., Reshetilova, T.A., Sakharovskii, V.G., Baskunov, B.P., and Seleznev, S.G., New Metabolites of Penicillium glandicola var. glandicola: Glandicolin A and Glandicolin B, Prikl. Biokhim. Mikrobiol., 1994, vol. 30, no. 3, pp. 410–414.Google Scholar
  15. 15.
    Kozlovskii, A.G., Solov'eva, T.F., Sakharovskii, V.G., and Adanin, V.M., Biosynthesis of Abnormal Ergot Alkaloids by the Fungus Penicillium aurantio-virens, Dokl. Akad. Nauk SSSR, 1981, vol. 260, no. 1, pp. 230–233.Google Scholar
  16. 16.
    Kozlovskii, A.G., Vinokurova, N.G., and Adanin, V.M., Diketopiperazine Alkaloids of the Fungus Penicillium piscarium, Prikl. Biokhim. Mikrobiol., 2000, vol. 36, no. 3, pp. 271–275.Google Scholar
  17. 17.
    Reshetilova, T.A., Solov'eva, T.F., Baskunov, B.P., and Kozlovskii, A.G., Investigation of Alkaloid Formation in Some Fungi of the Genus Penicillium, Mikrobiologiya, 1992, vol. 61, no. 3, pp. 873–879.Google Scholar
  18. 18.
    Kozlovskii, A.G., Reshetilova, T.A., Medvedeva, T.N., Arinbasarov, M.U., Sakharovskii, V.G., and Adanin, V.M., Intra-and Extracellular Alkaloids of Penicillium roquefortii, Biokhimiya, 1979, vol. 44, no. 9, pp. 1691–1700.Google Scholar
  19. 19.
    Solov'eva, T.F., Baskunov, B.P., and Reshetilova, T.A., Puberulin A, a New Diketopiperazine Alkaloid of the Fungus Penicillium rugulosum Thom VKM F-352, Prikl. Biokhim. Mikrobiol., 1997, vol. 33, no. 1, pp. 66–69.Google Scholar
  20. 20.
    Solov'eva, T.F., Reshetilova, T.A., Baskunov, B.P., Grigoryan, K.M., and Kozlovskii, A.G., Alkaloids of the Fungi of the Genus Penicillium Isolated from Food Stuffs, Prikl. Biokhim. Mikrobiol., 1995, vol. 31, no. 5, pp. 545–550.Google Scholar
  21. 21.
    Vinokurova, N.G., Ozerskaya, S.M., Zhelifonova, V.P., and Adanin, V.M., Taxonomic Position and Nitrogen-Containing Secondary Metabolites of Penicillium vitale Pidoplichko et Bilai apud Bilai, Mikrobiologiya, 2000, vol. 69, no. 3, pp. 415–419.Google Scholar
  22. 22.
    Kozlovskii, A.G., Vinokurova, N.G., and Zhelifonova, V.P., Mycotoxin Production Profiles of Penicillium vulpinum (Cooke & Massee) Seifert & Samson Strains, Mikrobiologiya, 2000, vol. 69, no. 1, pp. 45–48.Google Scholar

Copyright information

© MAIK “Nauka/Interperiodica” 2001

Authors and Affiliations

  • L. V. Boichenko
    • 1
  • D. M. Boichenko
    • 1
  • N. G. Vinokurova
    • 1
  • T. A. Reshetilova
    • 1
  • M. U. Arinbasarov
    • 1
  1. 1.Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of SciencesPushchino, Moscow oblastRussia

Personalised recommendations