Roles of three Fusarium graminearum membrane Ca²⁺ channels in the formation of Ca²⁺ signatures, growth, development, pathogenicity and mycotoxin production

Similar to animals and plants, external stimuli cause dynamic spatial and temporal changes of cytoplasmic Ca²⁺ in fungi. Such changes are referred as the Ca²⁺ signature and control cellular responses by modulating the activity or location of diverse Ca²⁺-binding proteins (CBPs) and also indirectly affecting proteins that interact with CBPs. To understand the mechanism underpinning Ca²⁺ signaling, therefore, characterization of how Ca²⁺ moves to and from the cytoplasm to create Ca²⁺ signatures under different conditions is fundamental. Three genes encoding plasma membrane Ca²⁺ channels in a Fusarium graminearum strain that expresses a fluorescent protein-based Ca²⁺ indicator in the cytoplasm were mutagenized to investigate their roles in the generation of Ca²⁺ signatures under different growth conditions and genetic backgrounds. The genes disrupted include CCH1 and MID1, which encode a high affinity Ca²⁺ uptake system, and FIG1, encoding a low affinity Ca²⁺ channel. Resulting mutants were also analyzed for growth, development, pathogenicity and mycotoxin production to determine how loss of each of the genes alters these traits. To investigate whether individual genes influence the function and expression of other genes, phenotypes and Ca²⁺ signatures of their double and triple mutants, as well as their expression patterns, were analyzed.