Protein interaction and gene expression analyses of SNF1 kinase complex components and its activating kinases in <i>M. oryzae</i>.

<p>(<b>A</b>) Different composition of the heterotrimeric SNF1 kinase complex and upstream kinases between <i>S. cerevisiae</i> and <i>M. oryzae</i>. (<b>B</b>) MoSnf1, MoSip2, and MoSnf4 interacted with each other, while no interaction was observed between MoSnf1 and its activating kinases in yeast two-hybrid assay. Yeast transformants expressing MoSnf1 plus MoSip2, MoSnf1 plus MoSnf4, MoSip2 plus MoSnf4, MoSnf1 plus MoSak1, or MoSnf1 plus MoTos3 were 10-fold serially diluted with a starter culture of 10⁶ cells/ml and then spotted (5 µl) onto SD-Trp-Leu-His-Ade medium. (<b>C</b>) Gene expression profiles of <i>MoSNF1</i>, <i>MoSIP2</i>, <i>MoSNF4</i>, <i>MoSAK1</i>, and <i>MoTOS3</i> among different developmental stages. Tested fungal tissues included vegetative hyphae (VH), conidia (CO), appressoria 8 hpi (AP), and invasive hyphae (72 hpi), which were within infected plant leaves (IP). Gene expression data, obtained from quantitative RT-PCR analysis, were normalized by using β-tubulin as an internal control and calibrated against the transcript abundances of VH stage.