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

钙离子（Ca2+）是动物、植物和真菌中重要的第二信使，但其在真菌感染过程中的作用知之甚少。本项目的目的是阐明钙离子信号在建立兼容的相互作用的禾生炭疽菌的作用。我们将通过对C.草生菌基因组序列。使用靶向缺失方法，我们将敲除已鉴定的候选基因，并确定敲除对真菌渗透前和渗透后发育的影响。在C.将通过用密码子优化的水母发光蛋白Ca2+报道基因转化真菌来鉴定草间生属感染过程。使用发光测定法和光子成像，我们将确定在什么阶段的相互作用，并在哪些细胞类型的Ca2+信号发生。将确定候选基因的缺失对那些Ca2+信号的影响。为了使识别的Ca2+信号具有更高的时空分辨率，我们将采用Ca2+依赖的cameleon荧光的比率成像。在Ca2+信号传导和致病性中发挥作用的蛋白质的功能将通过亚细胞定位和酵母中的异源表达进一步检查。在这个项目中获得的知识不仅将促进我们对Ca2+依赖性过程的理解，从而形成相容的相互作用，而且还将为抗真菌剂的开发提供新的靶点。

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.