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Calcium signals and associated genes
Project
Project code: DFG FOR 666
Contract period: 01.01.2006
- 31.12.2010
Purpose of research: Basic research
The aim of this project is to elucidate the role of Ca2+ signalling processes in C. graminicola during the establishment of a compatible interaction, including germination, appressorium formation, and subsequent biotrophic and necrotrophic growth. We will identify candidate genes for upstream and downstream components of the Ca2+ signal (i.e. Ca2+ channels, transporters, and sensor proteins) by bioinformatics analysis supported by project B7 of the C. graminicola genome sequence traces. Studies on other filamentous fungi and yeast have revealed a number of genes directly involved in Ca2+ signalling (Peiter et al. 2005, Peiter et al. 2007) that will serve as candidates in the present proposal. Using a targeted deletion approach we will knock out identified candidate genes and determine the effect of knock-out on fungal performance, at the various interaction stages. To identify individual Ca2+ signals in the C. graminicola infection process, the fungus has been transformed with the novel codon-optimized Ca2+ reporter protein aequorinS. Using luminometry and photon imaging, we will determine at what stages and in which cell types Ca2+ signals occur (e.g., spore germination, appressorium formation, switch from biotrophy to necrotrophy), as well as the kinetics of the Ca2+ transients. The aequorin method will subsequently be used to determine the effect of gene deletions on those Ca2+ signals. In parallel, we will attempt to establish a ratiometric Ca2+ imaging approach based on the cameleon YC3.6 reporter. This will help to resolve Ca2+ signals with a greatly increased temporal and spatial resolution. For proteins displaying a role in Ca2+ signalling and pathogenicity, the function will be further elucidated by heterologous expression of their full-length cDNAs in the respective yeast mutants. The combination of experimental approaches in this project will help to identify Ca2+ signalosome components involved in compatible interactions, to identify their roles in Ca2+ signalling events, and to link those events with the infection process. This knowledge will not only advance our understanding of the signalling processes leading to the formation of a compatible biotrophic interaction, but will also provide targets for the development novel agents with fungicidal action.
Section overview
Subjects
- Crop Protection
Collaborative Project
FOR 666: Mechanisms of compatibility: Reprogramming of plant metabolism by fungal effector molecules
