Biotecnología Vegetal (Oct 2013)

Quantification <i>of Mycosphaerella fijiensis</i> <i>in vitro</i> growth by absorbance readings

  • Mileidy Cruz-Martín,
  • Mayra Acosta-Suárez,
  • Eilyn Mena,
  • Berkis Roque,
  • Michel Leiva-Mora,
  • Tatiana Pichardo,
  • Rosa del Pilar Castro,
  • Yelenys Alvarado-Capó

Journal volume & issue
Vol. 13, no. 4

Abstract

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The slow growth of Mycosphaerella fijiensis Morelet in synthetic and semi-synthetic media as well as the characteristics of their colonies makes the use of traditional methods for evaluating the fungal growth are inadequate and impractical. Therefore, it requires a method to quantify growth in small volumes of substance and in short periods of time. The aim of this study was to quantify the M. fijiensis growth, through absorbance readings on the time, of mycelial suspensions obtained from cultures in liquid medium. Mycelial dry weight of the pathogen grown in liquid culture in flasks of 250 ml capacity and the absorbance in 96 well plates was compared. Similar procedure was used to evaluate the effect of bacterial culture filtrates with in vitro antifungal activity against M. fijiensis. A linear relationship between absorbance at 595 nm of fungal growth in liquid culture medium with respect to the dry mass was found. Absorbance readings using 96-well plates were possible to quantify the M. fijiensis mycelial suspensions growth in liquid culture. Also, there may be applied to assess the growth inhibition of the pathogen for bacterial culture filtrates with results at 48 h incubation. Key words: antifungal activity assay, biomass, DO595