Skip to main content
SpringerLink
Log in

In Vitro Production of Biotrophic-Like Cultures of Crinipellis perniciosa, the Causal Agent of Witches’ Broom Disease of Theobroma cacao

  • Published:
Current Microbiology Aims and scope Submit manuscript

Abstract

Witches’ broom disease (WBD) of cacao, caused by the hemibiotrophic fungus, Crinipellis perniciosa, exhibits a succession of symptoms that are caused by the biotrophic phase of the fungus. However, the study of this biotrophic phase is limited by its exclusive growth inside the plant or in the presence of callus. Here we report for the first time a method for the growth and maintenance of the biotrophic-like phase of C. perniciosa on a defined medium with metabolites found in the diseased tissues. Our results suggest that glycerol is a key carbon source for this interaction. This is a crucial achievement toward understanding the biology of this fungus during the infectious phase of WBD.

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

Fig. 1
Fig. 2

Similar content being viewed by others

Literature Cited

  1. Arruda MCC, Ferreira MASV, Miller RNG, Resende MLV, Felipe MSS (2003) Nuclear and mitochrondrial rDNA variability in Crinipellis perniciosa from different geographic origins and hosts. Mycol Res 107:25–37

    PubMed  Google Scholar 

  2. Calle H, Cook AA, Fernando SY (1982) Histology of witches broom caused in cacao by Crinipellis perniciosa. Phytopathology 72:1479–1481

    Google Scholar 

  3. Brownlee HE, Hedger J, Scott IM (1990) Host extracts cause morphological variation in germ tubes of the cocoa pathogen, Crinipellis perniciosa. Mycol Res 94:543–547

    Google Scholar 

  4. Brownlee HE, Hedger J, Scott IM (1992) Effects of a range of procyanidins on the cocoa pathogen Crinipellis perniciosa. Physiol Mol Plant Pathol 40:227–232

    Article  CAS  Google Scholar 

  5. Brownlee HE, Mceun AR, Hedger J, Scott IM (1990) Antifungal effects of cocoa tannin on the witches’ broom pathogen Crinipellis perniciosa. Physiol Mol Plant Pathol 36:39–48

    Article  CAS  Google Scholar 

  6. Delgado JC, Cook AA (1976) Nuclear condition of basidia, basidiospores and mycelia of Marasmius perniciosa. Can J Bot 54:66–72

    Google Scholar 

  7. Evans HC (1980) Pleomorphism in Crinipellis perniciosa, causal agent of Witches’ broom disease of cocoa. T Brit Mycol Soc 74:515–526

    Google Scholar 

  8. Frias GA, Purdy LH (1991) Infection biology of Crinipellis perniciosa on vegetative flushes of cacao. Plant Dis 75:552–556

    Google Scholar 

  9. Frias GA, Purdy LH (1995) An inoculation method for evaluating resistance of cacao to Crinipelis perniciosa. Plant Dis 79:787–791

    Google Scholar 

  10. Goodwin PH, Chen GYJ (2002) High expression of a sucrose non–fermenting (SNF1)-related protein kinase from Colletotrichum gloeosporoides f. sp. malvae is associated with penetration of Malva pusilla FEMS Microbiol Lett 215:169–174

    CAS  PubMed  Google Scholar 

  11. Griffith GW (2004) Witches’ brooms and frosty pods: Biologist 51:71–75

    Google Scholar 

  12. Griffith GW, Hedger JN (1993) A novel method for producing basidiocarps of the cocoa pathogen Crinipellis perniciosa using a bran-vermiculite medium. Neth J Plant Pathol 99:227–230

    Article  Google Scholar 

  13. Griffith GW, Hedger JN (1994a). The breeding biology of biotypes of the witches’ broom pathogen, Crinipellis perniciosa. Heredity 72:278–289

    Google Scholar 

  14. Griffith GW, Hedger JN (1994b). Dual culture of Crinipellis perniciosa and potato callus. Eur J Plant Pathol 100:371–379

    Article  Google Scholar 

  15. Hedger JN, Pickering V, Aragundi JA (1987) Variability of populations of the witches’ broom disease of cocoa (Crinipellis perniciosa). T Br Mycol Soc 88:533–546

    Google Scholar 

  16. Kilaru A Hasenstein KH (2005) Development and pathogenicity of the fungus Crinipellis perniciosa on interaction with cacao leaves. Phytopathology 95:101–107

    Google Scholar 

  17. Klose J, Sá MM, Kronstad JW (2004) Lipid-induced filamentous growth in Ustilago maydis. Mol Microbiol 52:823–835

    Article  CAS  PubMed  Google Scholar 

  18. Mandel M, Parrish FW, Reese ET (1962) Sophorose as an inhibitor of cellulase in Trichoderma viride. J Bacteriol 83:400–408

    Google Scholar 

  19. Meinhardt LW, Bellato CM, Tsai SM (2001) SYBR green I used to evaluate the nuclei number of fungal mycelia. BioTechniques 31:42–46

    CAS  PubMed  Google Scholar 

  20. Meinhardt LW, Wulff NA, Bellato CM, Tsai SM (2002a) Genetic analyses of Rhizoctonia solani isolates from Phaseolus vulgaris grown in the Atlantic rainforest region of São Paulo, Brazil. Fitopatol Bras 27:259–267

    CAS  Google Scholar 

  21. Meinhardt LW, Wulff NA, Bellato CM, Tsai SM (2002b) Telomere and microsatellite primers reveal diversity among Sclerotinia sclerotiorum isolates from Brazil. Fitopatol Bras 27:211–215

    CAS  Google Scholar 

  22. Orchard JE, Hardwick K (1988) Photosynthesis, carbohydrate translocation and metabolism of host and fungal tissues in cacao seedlings infected with Crinipellis perniciosa. In: 10th International Cocoa Research Conference. Cocoa Producers’ Alliance, Lagos, Nigeria, pp 325–330

  23. Purdy LH, Schmidt RA (1996) Status of cacao witches’ broom: Biology, epidemiology and management. Ann Rev Phytopathol 34:573–594

    CAS  Google Scholar 

  24. Resende MLV, Gutemberg BAN, Silva LHCP, Niella GR, Carvalho GA, Santiago DVR, et al (2000) Crinipellis perniciosa proveniente de um novo hospedeiro, Heteropterys acutifolia, é patogênico a T. cacao. Fitopatol Bras 25:88–91

    Google Scholar 

  25. Rincones J, Meinhardt LW, Vidal BC, Pereira GAG (2003) Electrophoretic karyotype analysis of Crinipellis perniciosa, the causal agent of witches’ broom disease of Theobroma cacao. Mycol Res 107:452–458

    Article  CAS  PubMed  Google Scholar 

  26. Scarpari LM, Meinhardt LW, Pereira GAG, et al (2005) Biochemical changes during the development of witches’ broom: The most important disease of coca in Brazil caused by Crinipellis perniciosa. J Exp Bot 56:865–877

    Article  CAS  PubMed  Google Scholar 

  27. Wei Y, Shen W, Dauk M, Wang F, Selvaraj G, Zou J (2004) Targeted gene disruption of glycerol-3-phosphate dehydrogenase in Colletotrichum gloeosporioides reveals evidence that glycerol is a significant transferred nutrient from host plant to fungal pathogen. J Biol Chem 276:429–435

    Google Scholar 

  28. White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TH (eds) PCR protocols: A guide to methods and applications. 1st ed. San Diego, CA: Academic, pp. 315–322

    Google Scholar 

Download references

Acknowledgments

The investigators thank the ongoing corporate support given by Almirante Cacau Agrícola (Masterfoods) and Cargill. This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (Nos. 00/10545-4 and 02/09280-1); Conselho Nacional de Pesquisa e Desenvolvimento (No. 68.0032/01-0; 47.1609/03-0); and Secretaria de Agricultura do Estado da Bahia.

Author information

Authors and Affiliations

  1. Laboratório de Genômica e Expressão, Departamento de Genética e Evolução, Instituto de Biologia, Universidade Estadual de Campinas, CP 6109, Campinas, São Paulo, 13083-970, Brazil

    Lyndel W. Meinhardt, Johana Rincones & Gonçalo A. G. Pereira

  2. Laboratório de Biologia Celular e Molecular, Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, Piracicaba, São Paulo, 13416-000, Brazil

    Cláudia de M. Bellato

  3. Departamento de Genética, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, São Paulo, 13400-970, Brazil

    Ricardo A. Azevedo

  4. Laboratório de Genômica e Expressão Gênica, Universidade Estados Santa Cruz, Rodovia Ilhéus-Itabuna, Km 16, Ilhéus, Bahia, 45650-000, Brazil

    Julio C. M. Cascardo

Authors
  1. Lyndel W. Meinhardt
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Cláudia de M. Bellato
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Johana Rincones
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ricardo A. Azevedo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Julio C. M. Cascardo
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gonçalo A. G. Pereira
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lyndel W. Meinhardt.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Meinhardt, L.W., Bellato, C.d., Rincones, J. et al. In Vitro Production of Biotrophic-Like Cultures of Crinipellis perniciosa, the Causal Agent of Witches’ Broom Disease of Theobroma cacao. Curr Microbiol 52, 191–196 (2006). https://doi.org/10.1007/s00284-005-0182-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00284-005-0182-z

Keywords

Search

Navigation