Skip to main content
SpringerLink
Log in

Reduction of the number of nuclei per cell ofHelminthosporium euphorbiae by protoplast isolation

  • Original Papers
  • Published:
Mycoscience

Abstract

Helminthosporium euphorbiae is a pathogen of the weedEuphorbia heterophylla, which causes severe losses in soybean (Glycine max) crops. The fungus causes leaf loss and affects germination, making it a promising biocontrol agent for this weed. In order to start a breeding program for this species, four isolates were examined for number of nuclei in the conidia and hyphae and nuclear behavior at different cultivation stages. The conidia were multinucleated with about 20 nuclei per conidium, and 5 to 7 nuclei were observed in the hyphae compartments. The high number of nuclei makes the genetic manipulation of this species diffucult, so the protoplast formation is an alternative for obtaining cells with a reduced number of nuclei. Thus the experimental conditions for the production and regeneration of protoplasts inH. euphorbiae were determined by assessing three enzymatic complexes and seven osmotic stabilizers. The efficiency of formation and regeneration frequency of the protoplasts varied depending on isolates, stabilizers and enzyme mixture used. The number of nuclei estimated per protoplast was reduced to 1 to 6, depending on the stage of mycelial growth during the protoplast formation process.

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

  • Bagagli, E., Valadares, M. C. C. and Azevedo, J. L. 1991. Parameiosis in the entomopathogenic fungusMetarhizium anisopliae (Metsch.) Sorokin. Rev. Brasil. Genet.14: 261–271.

    Google Scholar 

  • Barcellos, F. G. 1993. Estudos básicos visando o melhoramento genético da linhagem Ma12 deMetarhizium anisopliae para esporulação. Monografia-Bacharelado em Ciências Biológicas-Universidade Estadual de Londrina.

  • Cerdeira, A. L. and Voll, E. 1981. Controle de amendoim bravo (Euphorbia heterophylla L.) através de herbicidas pósemergentes. Seminário Nacional de Pesquisa de Soja. EMBRAPA-CNPSo, Brasília1: 311–317.

    Google Scholar 

  • Davis, B. 1985. Factors influencing protoplast isolation. In: Fungal Protoplast, Applications in Biochemistry and Genetics, (ed. by Peberdy, J. F. and Ferenczy, L., Marcel Dekker, New York.

    Google Scholar 

  • Gazziero, D. L. P. 1985. Controle químico deEuphorbia heterophylla. Resultados de Pesquisa da Soja 1984/85. EMBRAPA-CNPSo, Londrina1: 335–338.

    Google Scholar 

  • Gazziero, D. L. P., Calcavara, P. R., Yorinori, J. T. and Arrabal, C. A. 1989. Avaliação da compatibilidade de misturas deHelminthosporium sp. com o herbicida pós-emergente Chlorimuron-ethyl, no controle de amendoim-bravo (Euphorbia heterophylla). Resultados de Pesquisa da Soja 1988/89. EMBRAPA-CNPSo, Londrina1: 279.

    Google Scholar 

  • Gazziero, D. L. P., Sarlo, S. M. Z. and Yorinori, J. T. 1993. Fatores que interferem na eficiência do fungoHelminthosporium sp. no controle biológico de amendoim-bravo (Euphorbia heterophylla). Resultados de Pesquisa da Soja 1989/90. EMBRAPA-CNPSo, Londrina1: 367–375.

    Google Scholar 

  • Gazziero, D. L. P. and Yorinori, J. T. 1987. Efeitos da hora de aplicação do fungoHelminthosporium sp. no controle de amendoim-bravo (Euphorbia heterophylla). Resultados de Pesquisa da Soja 1985/86. EMBRAPA-CNPSo, Londrina1: 395–401.

    Google Scholar 

  • Gazziero, D. L. P. and Yorinori, J. T. 1988. Avaliação da compatibilidade da mistura deHelminthosporium sp. com herbicidas pós-emergentes no controle de amendoim-bravo (Euphorbia heterophylla). Resultados de Pesquisa da Soja 1986/87. EMBRAPA-CNPSo, Londrina1: 336–338.

    Google Scholar 

  • Hamlym, P. F., Bradshaw, R. E., Mellon, F. W., Santiago, C. M., Wilson, J. M. and Peberdy, J. F. 1981. Efficient protoplast isolation from fungi using commercial enzymes. Enzyme Microb. Technol.3: 321–325.

    Article  Google Scholar 

  • Iijima, Y. and Yanagi, S. O. 1986. A method for the high yield preparation of and high frequency regeneration of BasidiomycetePleurotus ostreatus (“Hiratake”) protoplasts using sulfite pulp waste components. Agric. Biol. Chem.50: 1855–1861.

    Google Scholar 

  • Knox-Davies, P. S. and Dickson, J. G. 1960. Cytology ofHelminthosporium turcium and its ascigerous stage,Trichometasphaeria turcica. Am. J. Bot.47: 328–339.

    Article  Google Scholar 

  • Leonard, R. J. 1988.Setosphaeria turcica, cause of northern corn leafblight, and otherSetosphaeria spp, Adv. Plant Pathol.6: 241–248.

    Google Scholar 

  • Muchovej, J. J. and Carvalho, A. O. 1989. A new combination forHelminthosporium euphorbiae. Mycotaxon25: 159–162.

    Google Scholar 

  • Peberdy, J. F. 1978. Protoplasts and their development. In: The filamentous fungi-Developmental mycology, (ed. by Smith, J. E. and Berry, D. R.), pp. 119–131. E. Arnold, London.

    Google Scholar 

  • Souza, C. G. M. 1992. Avaliação de isolados deHelminthosporium sp. quanto à produção de fitotoxina para o controle biológico deE. heterophylla. Monografia-Bacharelado, Universidade Estadual de Londrina.

  • Tanaka, Y., Murata, N. and Kato, H. 1979. Behavior of nuclei and chromosomes during ascus development in the mating between either rice-strain or weeping lovegrass-strain and ragi-strain ofPyricularia. Ann. Phytopathol. Soc. Japan45: 182–191.

    Google Scholar 

  • Yorinori, J. T. 1985. Biological control of milk weed (E. heterophylla) with pathogenic fungi. In: International Symposium on Biological Control of Weeds, (ed. by Delfosse, E. S.), pp. 677–681. Vancouver, Canada.

  • Yorinori, J. T., Assis, R. and Gazziero, D. L. P. 1985a. Avaliação da compatibilidade de misturas deHelminthosporium sp. com inseticidas e herbicidas e o efeito no controle deE. heterophylla. Resultados de Pesquisa de Soja 1984/85. EMBRAPA-CNPSo, Londrina1: 320–322.

    Google Scholar 

  • Yorinori, J. T., Assis, R. and Gazziero, D. L. P. 1985b. Avaliação da longevidade de esporos deHelminthosporium sp. e produção massal de inóculo. Resultados de Pesquisa de Soja 1984/85. EMBRAPA-CNPSo, Londrina1: 325–326.

    Google Scholar 

  • Yorinori, J. T. 1987. Controle biológico de ervas daninhas com microrganismos. Reunião sobre Controle Biológico e Doenças de Plantas. Campinas1: 20–35.

    Google Scholar 

  • Yorinori, J. T. and Gazziero, D. L. P. 1987. Avaliação da Iongevidade de esporos deHelminthosporium sp. Resultados de Pesquisa de Soja 1985/86. EMBRAPA-CNPSo, Londrina1: 402–403.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Departmento de Biologia Geral-Centro de Ciências Biológicas-Universidade Estadual de Londrina, Campus Universitário, Caixa Postal 6001, CEP 86051-970, Londrina, Pr. Brasil

    Tânia Aparecida da Silva & Luzia Doretto Paccola-Meirelles

Authors
  1. Tânia Aparecida da Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Luzia Doretto Paccola-Meirelles
    View author publications

    You can also search for this author in PubMed Google Scholar

About this article

Cite this article

Aparecida da Silva, T., Paccola-Meirelles, L.D. Reduction of the number of nuclei per cell ofHelminthosporium euphorbiae by protoplast isolation. Mycoscience 42, 313–320 (2001). https://doi.org/10.1007/BF02461212

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02461212

Key words

Search

Navigation