The role of calcium signalling in the establishment of compatible Colletotrichum graminicola interactions

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Calcium (Ca2+) is a pivotal second messenger in animals, plants, and fungi, but its role in the fungal infection process is poorly understood. The aim of this project is to elucidate the role of Ca2+ signalling in the establishment of compatible interactions by Colletotrichum graminicola. We will identify candidate genes for components of the Ca2+ signalling machinery by bioinformatic analysis of the C. graminicola genome sequence. Using a targeted deletion approach, we will knock out identified candidate genes and determine the effect of knock out on fungal pre- and post-penetration development. Individual Ca2+ signals in the C. graminicola infection process will be identified by transforming the fungus with a codon-optimised aequorin Ca2+reporter. Using luminometry and photon imaging, we will determine at what stages of the interaction and in which cell types Ca2+ signals occur. Effects of the deletion of candidate genes on those Ca2+ signals will be determined. To characterise the identified Ca2+ signals with an increased spatio-temporal resolution, we will employ the ratiometric imaging of Ca2+-dependent cameleon fluorescence. The function of proteins displaying a role in Ca2+signalling and pathogenicity will be further examined by subcellular localisation and heterologous expression in yeast. The knowledge gained in this project will not only advance our understanding of Ca2+-dependent processes leading to the formation of a compatible interaction, but will also provide novel targets for the development of antifungal agents.