European Journal of Plant Pathology

, Volume 102, Issue 9, pp 883–889 | Cite as

The use of ergosterol in the pathogenic fungusBipolaris sorokiniana for resistance rating of barley cultivars

  • Torsten Gunnarsson
  • Ingrid Almgren
  • Pernilla Lydén
  • Helena ékesson
  • Hans -Börje Jansson
  • Göran Odham
  • Mats Gustafsson
Research Articles

Abstract

Ergosterol content in the plant pathogenic fungusBipolaris sorokiniana was determined in different matrices including mycelium, spores, culture filtrate and infected barley leaves. Ergosterol was extracted with methanol, hydrolysed with KOH and quantified by reverse phase high performance liquid chromatography (HPLC). Our procedure was used to study how the ergosterol concentration ofB. sorokiniana varied due to fungal age and nutrient availability when growing in liquid medium. It was found that the ergosterol content decreased with fungal age. The decrease was not due to leakage. It was also found that a change to a less nutrient-rich medium caused an increase in ergosterol content whereas a change to a rich medium led to a decrease. The procedure was also used for quantification of fungal infections in complex matrices (e.g. leaves). The development of fungal infection in barley leaves was followed during 10 days. Visual grading of leaf spots was also compared to ergosterol content in three varieties of barley. The ergosterol content in the leaves increased exponentially until day 7, and the grading of the leaf spots was correlated to the ergosterol content. Our results show that, despite a great variation, ergosterol may be used as a biomarker to detect and quantify fungal infections in a given matrix.

Key words

fungal pathogen Hordeum vulgare HPLC HPLC-MS 

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References

  1. Carlson H, Nilsson P, Jansson H-B and Odham G (1991) Characterization and determination of prehelminthosporal, a toxin from the plant pathogenic fungusBipolaris sorokiniana, using liquid chromatography/mass spectrometry. J Microbiol Meth 13: 259–269Google Scholar
  2. Davis MW and Lamar RT (1992) Evaluation of methods to extract ergosterol for quantification of soil fungal biomass. Soil Biol Biochem 24(3): 189–198Google Scholar
  3. Gessner MO, Bauchrowitz MA and Escautier M (1991) Extraction and quantification of ergosterol as a measure of fungal biomass in leaf litter. Microb Ecol 22: 285–291Google Scholar
  4. Gessner MO and Chauvet E (1993) Ergosterol-to-biomass conversion factors for aquatic hyphomycetes. Appl Environm Microbiol 59: 502–507Google Scholar
  5. Gordon TR and Webster RK (1984) Evaluation of ergosterol as an indicator of infestation of barley seed byDrechslera graminea. Phytopathol 74: 1125–1127Google Scholar
  6. Gordon TR and Webster RK (1986) Identification of ergosterol as a metabolite ofDrechslera graminea andD. teres. Can J Microbiol 32: 69–71Google Scholar
  7. Gretenkort MA and Ingram DS (1993) The use of ergosterol as a quantitative measure of the resistance of cultured tissues ofBrassica napus ssp.oleifera toLeptosphaeria maculans. J Phytopathol 138: 217–224Google Scholar
  8. Griffiths HM, Jones DG and Akers A (1985) A bioassay for predicting the resistance of wheat leaves toSeptoria nodorum. Ann Appl Biol 107: 293–300Google Scholar
  9. Kurppa A (1985) Reactions of spring barley cultivars grown in Finland to soil-borne infection byBipolaris sorokiniana and to its toxic metabolites. J Agric Sci Finl 57: 85–96Google Scholar
  10. Liljeroth E, Jansson H-B and SchÄfer W (1993) Transformation ofBipolaris sorokiniana with the GUS-gene and the use for studying fungal colonization of barley roots. Phytopathol 84: 1484–1489Google Scholar
  11. Matcham SE, Jordan BR and Wood DA (1985) Estimation of fungal biomass in a solid substrate by three independent methods. Appl Microbiol Biotechnol 21: 108–112Google Scholar
  12. Martin F, Delaruelle C and Hilbert J-L (1990) An improved ergosterol assay to estimate the fungal biomass in ectomycorrhizas. Mycol Res 94: 1059–1064Google Scholar
  13. Newell SY, Arsuffi TL and Fallon RD (1988) Fundamental procedures for determining ergosterol content of decaying plant material by liquid chromatography. Appl Environm Microbiol 54: 1876–1879Google Scholar
  14. Nout MJR, Bonants-van Laarhoven TMG, de Jong P and de Koster PG (1987) Ergosterol content ofRhizopus oligisporus NRRL 5905 grown in liquid and solid substrates. Appl Microbiol Biotechnol 26: 456–461Google Scholar
  15. Piening LJ (1973) Differential yield response to ten barley cultivars to common root rot. Can J Plant Sci 53: 763–764Google Scholar
  16. Salmanowicz B and Nylund J-E (1988) High performance chromatography determination of ergosterol as a measure of ectomycorrhiza infection in Scots pine. Eur J For Pathol 18: 291–298Google Scholar
  17. Schmitz O, Danneberg G, Hundeshagen B, Klingner A and Both H (1991) Quantification of Vesicular-Arbuscular mycorrhiza by biochemical parameters. J Plant Physiol 139: 106–114Google Scholar
  18. Seitz LM, Mohr HF, Burroughs R and Sauer DB (1977) Ergosterol as an indicator of fungal invasion in grains. Cereal Chem 54: 1207–1217Google Scholar
  19. Seitz LM, Sauer DB, Burroughs R, Mohr HE and Hubbard JD (1979) Ergosterol as a measure of fungal growth. Phytopathol 358: 1202–1203Google Scholar
  20. Skoropad WP and Arny DC (1957) The influence of amino acids on the growth of two strains ofHelminthosporium gramineum. Phytopathol 47: 249–252Google Scholar
  21. Snijders CHA and Krechting CF (1992) Inhibition of deoxynivalenol translocation and fungal colonization inFusarium head blight resistant wheat. Can J Bot 70: 1570–1576Google Scholar
  22. Stack RW (1982) Yield losses in spring barley due to common root rot in eastern North Dakota. Phytopathl 72: 1139–1140Google Scholar
  23. Torres M, Viladrich R, Sanchis V and Canela R (1992) Influence of age on ergosterol content in mycelium ofAspergillus ochraceus. Lett Appl Microbiol 15: 20–22Google Scholar
  24. West AW and Grant WD (1987) Use of ergosterol, diaminopimelic acid and glucosamine contents of soils to monitor changes in microbial populations. Soil Biol Biochem 19: 607–612Google Scholar
  25. Young JC and Games DE (1993) Supercritical fluid extraction and supercritical fluid chromatography of the fungal metabolite ergosterol. J Agric Food Chem 41: 577–581Google Scholar

Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Torsten Gunnarsson
    • 1
  • Ingrid Almgren
    • 2
  • Pernilla Lydén
    • 1
  • Helena ékesson
    • 3
  • Hans -Börje Jansson
    • 3
  • Göran Odham
    • 1
  • Mats Gustafsson
    • 2
  1. 1.Chemical Ecology and Ectoxicology, Dept. of EcologyLund UniversityLundSweden
  2. 2.Dept. of Plant Breeding ResearchSwedish Univ. of Agr. Sci.SvalövSweden
  3. 3.Microbial Ecology, Dept. of EcologyLund UniversityLundSweden

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