Fungal Growth and Transformation Protocols

The wild type M. oryzae strain Guy11 and its derivative strains were grown on complete medium (CM) plates [20 (link)] at 28°C. Conidia were harvested from 9-day-old cultures grown on CM plates. Genomic DNA and total RNA were extracted from 3-day-old cultures grown in liquid CM. Fungal transformants were screened in CM plates supplemented with 200 μg ml⁻¹ of hygromycin B (Roche Applied Science, Mannheim, Germany) or 250 μg ml⁻¹ of glufosinate ammonium (Sigma-Aldrich, St Louis, MO, USA), depending on the selection marker. Evaluations of conidiation and vegetative growth were performed as previously described [21 (link)]. And growth rate of M. oryzae was determined by measuring the colony diameter of 6-day-old cultures on CM plates supplemented with chemicals.