Skip to main content
SpringerLink
Log in

Production of Calcium Oxalate Crystals by the Basidiomycete Moniliophthora perniciosa, the Causal Agent of Witches’ Broom Disease of Cacao

  • Published:
Current Microbiology Aims and scope Submit manuscript

Abstract

Oxalic acid has been shown as a virulence factor for some phytopathogenic fungi, removing calcium from pectin and favoring plant cell wall degradation. Recently, it was published that calcium oxalate accumulates in infected cacao tissues during the progression of Witches’ Broom disease (WBD). In the present work we report that the hemibiotrophic basidiomycete Moniliophthora perniciosa, the causal agent of WBD, produces calcium oxalate crystals. These crystals were initially observed by polarized light microscopy of hyphae growing on a glass slide, apparently being secreted from the cells. The analysis was refined by Scanning electron microscopy and the compositon of the crystals was confirmed by energy-dispersive x-ray spectrometry. The production of oxalate by M. perniciosa was reinforced by the identification of a putative gene coding for oxaloacetate acetylhydrolase, which catalyzes the hydrolysis of oxaloacetate to oxalate and acetate. This gene was shown to be expressed in the biotrophic-like mycelia, which in planta occupy the intercellular middle-lamella space, a region filled with pectin. Taken together, our results suggest that oxalate production by M. perniciosa may play a role in the WBD pathogenesis mechanism.

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
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Akamatsu Y, Ohta A, Takahashi M, et al. (1991) Enzymatic formation of oxalate from oxaloacetate with cell-free-extracts of the brown-rot fungus Tyromyces palustris in relation to the biodegradation of cellulose. Mokuzai Gakkaishi 37:575–577

    CAS  Google Scholar 

  2. Bateman DF, Beer SV (1965) Simultaneous production and synergistic action of oxalic acid and polygalacturonase during pathogenesis by Sclerotium rolfsii. Phytopathology 55:204

    PubMed  CAS  Google Scholar 

  3. Ceita GO, Macêdo JNA, Santos TB, et al. (2007) Involvement of calcium oxalate degradation during programmed cell death in Theobroma cacao tissues triggered by the hemibiotrophic fungus Moniliophthora perniciosa. Plant Sci 173:106–117

    Article  CAS  Google Scholar 

  4. Cessna SG, Sears VE, Dickman MB, et al. (2000) Oxalic acid, a pathogenicity factor for Sclerotinia sclerotiorum, suppresses the oxidative burst of the host plant. Plant Cell 12:2191–2199

    Article  PubMed  CAS  Google Scholar 

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

    Google Scholar 

  6. Dias BBA, Cunha WG, Morais LS, et al. (2006) Expression of an oxalate decarboxylase gene from Flammulina sp. in transgenic lettuce (Lactuca sativa) plants and resistance to Sclerotinia sclerotiorum. Plant Pathol 55:187–193

    Article  CAS  Google Scholar 

  7. Dutton MV, Evans CS (1996) Oxalate production by fungi: its role in pathogenicity and ecology in the soil environment. Can J Microbiol 42:881–895

    Article  CAS  Google Scholar 

  8. Evans HC (1980) Pleomorphism in Crinipellis perniciosa, causal agent of witches broom disease of cocoa. Trans Br Mycol Soc 74:515–523

    Article  Google Scholar 

  9. Evans HC, Bastos CN (1980) Basidiospore germination as a means of assessing resistance to Crinipellis perniciosa (Witches Broom Disease) in cocoa cultivars. Trans Bri Mycol Soc 74:525–536

    Google Scholar 

  10. Franceschi V (2001) Calcium oxalate in plants. Trends Plant Sci 6:331–331

    Article  PubMed  CAS  Google Scholar 

  11. Garcia O, Macedo JA, Tiburcio R, et al. (2007) Characterization of necrosis and ethylene-inducing proteins (NEP) in the basidiomycete Moniliophthora perniciosa, the causal agent of witches’ broom in Theobroma cacao. Mycol Res 111:443–455

    Article  PubMed  CAS  Google Scholar 

  12. Gesteira A, Micheli F, Carels N, et al. (2007) Comparative analysis of expressed genes from cacao meristems infected by Moniliophthora perniciosa. Ann Bot 100:129–140

    Article  PubMed  CAS  Google Scholar 

  13. Goldstein J, Newbury D, Joy D, et al. (2003) Scanning electron microscopy and x-ray microanalysis. Vol 3. Springer Science+Business Media, New York

    Google Scholar 

  14. Griffith GW, Nicholson J, Nenninger A, et al. (2003) Witches’ brooms and frosty pods: two major pathogens of cacao. New Zeal J Bot 41:423–435

    Google Scholar 

  15. Guimaraes RL, Stotz HU (2004) Oxalate production by Sclerotinia sclerotiorum deregulates guard cells during infection. Plant Physiol 136:3703–3711

    Article  PubMed  CAS  Google Scholar 

  16. Han Y, Joosten HJ, Niu WL, et al. (2007) Oxaloacetate hydrolase, the C-C bond lyase of oxalate secreting fungi. J Biol Chem 282:9581–9590

    Article  PubMed  CAS  Google Scholar 

  17. Kesarwani M, Azam M, Natarajan K, et al. (2000) Oxalate decarboxylase from Collybia velutipes—molecular cloning and its overexpression to confer resistance to fungal infection in transgenic tobacco and tomato. J Biol Chem 275:7230–7238

    Article  PubMed  CAS  Google Scholar 

  18. Kubicek CP, Schreferlkunar G, Wohrer W, et al. (1988) Evidence for a cytoplasmic pathway of oxalate biosynthesis in Aspergillus niger. Appl Environ Microbiol 54:633–637

    PubMed  CAS  Google Scholar 

  19. Lenz H, Wunderwald P, Eggerer H (1976) Partial purification and some properties of oxalacetase from Aspergillus niger. Eur J Biochem 65:225–236

    Article  PubMed  CAS  Google Scholar 

  20. Maximova SN, Marelli JP, Young A, et al. (2006) Over-expression of a cacao class I chitinase gene in Theobroma cacao L. enhances resistance against the pathogen, Colletotrichum gloeosporioides. Planta 224:740–749

    Article  PubMed  CAS  Google Scholar 

  21. Meinhardt LW, Bellato CD, Tsai SM (2001) SYBR (R) Green I used to evaluate the nuclei number of fungal mycelia. Biotechniques 31:42–46

    PubMed  CAS  Google Scholar 

  22. Munir E, Yoon JJ, Tokimatsu T, et al. (2001) A physiological role for oxalic acid biosynthesis in the wood-rotting basidiomycete Fomitopsis palustris. Proc Natl Acad Sci USA 98:11126–11130

    Article  PubMed  CAS  Google Scholar 

  23. Pedersen H, Christensen B, Hjort C, et al. (2000) Construction and characterization of an oxalic acid nonproducing strain of Aspergillus niger. Metab Eng 2:34–41

    Article  PubMed  CAS  Google Scholar 

  24. Pennman D, Britton G, Hardwick K, et al. (2000) Chitin as a measure of biomass of Crinipellis perniciosa, causal agent of witches broom disease of cocoa. Mycol Res 104:671–675

    Article  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  28. Sexton AC, Cozijnsen AJ, Keniry A, et al. (2006) Comparison of transcription of multiple genes at three developmental stages of the plant pathogen Sclerotinia sclerotiorum. FEMS Microbiol Lett 258:150–160

    Article  PubMed  CAS  Google Scholar 

  29. Silva SDVM, Matsuoka K (1999) Histologia da interação Crinipellis perniciosa em cacaueiros suscetível e resistente à vassoura-de-bruxa. Fitopat Bras 24:54–59

    Google Scholar 

  30. Tokimatsu T, Nagai Y, Hattori T, et al. (1998) Purification and characteristics of a novel cytochrome c dependent glyoxylate dehydrogenase from a wood-destroying fungus Tyromyces palustris. FEBS Lett 437:117–121

    Article  PubMed  CAS  Google Scholar 

  31. Whitney KD, Arnott HJ (1986) Calcium oxalate crystals and basidiocarp dehiscence in Geastrum saccatum (Gasteromycetes). Mycologia 78:649–656

    Article  CAS  Google Scholar 

  32. Whitney KD, Arnott HJ (1987) Calcium oxalate crystal morphology and development in Agaricus bisporus. Mycologia 79:180–187

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico; No. 472279/2006-8) and FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo; Nos. 06/50794-0, 06/59843-3, and 07/51030-6). The authors would like to thank Daniel Bratfisch Razzo for operation of the scanning electron microscope (JSM 6360 LV), Rafaela F. Camargo for help in preparing fungal samples, Dr. Francisco Javier Medrano for critical suggestions, and Dr. Johana Rincones and Dr. Carol H. Collins for manuscript revision.

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, 13083-970, São Paulo, Brazil

    Maria Carolina S. do Rio, Bruno V. de Oliveira, Daniela P. T. de Tomazella & Gonçalo A. G. Pereira

  2. Departamento de Química Analítica, Instituto de Química, Universidade Estadual de Campinas, CP 6157, Campinas, 13083-970, São Paulo, Brazil

    José A. Fracassi da Silva

Authors
  1. Maria Carolina S. do Rio
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bruno V. de Oliveira
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Daniela P. T. de Tomazella
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. José A. Fracassi da Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Gonçalo A. G. Pereira
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gonçalo A. G. Pereira.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rio, M.C.S.d., de Oliveira, B.V., de Tomazella, D.P.T. et al. Production of Calcium Oxalate Crystals by the Basidiomycete Moniliophthora perniciosa, the Causal Agent of Witches’ Broom Disease of Cacao. Curr Microbiol 56, 363–370 (2008). https://doi.org/10.1007/s00284-007-9091-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00284-007-9091-7

Keywords

Search

Navigation