Fig. 3. Expression analysis of splaA and splaB.

(A) splaA and splaB expression levels were determined by RT-PCR analysis of total RNA extracted from A. oryzae RIB40 pre-cultured in DPY liquid medium for 24 h, transferred to the following fresh media, and grown for 4 h at 30 °C unless stated otherwise: (left to right) CD medium, CD without any carbon source, CD without any nitrogen source, CD adjusted to pH 8.0, CD medium at 4 °C, 15 °C and 40 °C, and CD containing 50 mM H₂O₂. γ-actin, utilized as an internal control, is shown at the bottom. (B) RT-PCR analysis of splaA and splaB during aerial hyphae and conidia formation. A. oryzae RIB40 was pre-cultured in DPY liquid medium for 24 h and then transferred to a Petri dish containing 10 ml of DPY liquid medium, so to allow the formation of aerial hyphae and conidia. After 6 h (early aerial hyphae formation), 18 h (completion of aerial hyphae formation and early conidiation), and 48 h (full conidiation), as revealed by microscopic examination, samples were collected and processed for RT-PCR as above (see ‘Materials and Methods’ for details).