Calcium oxalate crystals: an integral component of the Sclerotinia sclerotiorum/Brassica carinata pathosystem

PLoS One. 2015 Mar 27;10(3):e0122362. doi: 10.1371/journal.pone.0122362. eCollection 2015.

Abstract

Oxalic acid is an important virulence factor for disease caused by the fungal necrotrophic pathogen Sclerotinia sclerotiorum, yet calcium oxalate (CaOx) crystals have not been widely reported. B. carinata stems were infected with S. sclerotiorum and observed using light microscopy. Six hours post inoculation (hpi), CaOx crystals were evident on 46% of stem sections and by 72 hpi on 100%, demonstrating that the secretion of oxalic acid by S. sclerotiorum commences before hyphal penetration. This is the first time CaOx crystals have been reported on B. carinata infected with S. sclerotiorum. The shape of crystals varied as infection progressed. Long tetragonal rods were dominant 12 hpi (68% of crystal-containing samples), but by 72 hpi, 50% of stems displayed bipyramidal crystals, and only 23% had long rods. Scanning electron microscopy from 24 hpi revealed CaOx crystals in all samples, ranging from tiny irregular crystals (< 0.5 μm) to large (up to 40 μm) highly organized arrangements. Crystal morphology encompassed various forms, including tetragonal prisms, oval plates, crystal sand, and druses. Large conglomerates of CaOx crystals were observed in the hyphal mass 72 hpi and these are proposed as a strategy of the fungus to hold and detoxify Ca2+ions. The range of crystal morphologies suggests that S. sclerotiorum growth and infection controls the form taken by CaOx crystals.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ascomycota / chemistry
  • Ascomycota / metabolism
  • Ascomycota / pathogenicity*
  • Brassica / chemistry
  • Brassica / metabolism
  • Brassica / microbiology*
  • Calcium Oxalate / chemistry*
  • Calcium Oxalate / metabolism
  • Crystallization
  • Hyphae / chemistry
  • Hyphae / pathogenicity
  • Plant Diseases / microbiology

Substances

  • Calcium Oxalate

Grants and funding

This work was supported by the Australia Research Council and the Department of Agriculture and Food Western Australia (Project LP100200113, “Factors responsible for host resistance to the pathogen Sclerotinia sclerotiorum for developing effective disease management in vegetable Brassicas”, (http://www.arc.gov.au/)). MU gratefully acknowledges the financial assistance of an Australian Postgraduate Award. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.