ARS | Publication request: Functional Genomics and Proteomics Studies of F. Graminearum

Submitted to: The Canadian Workshop on Fusarium Head Blight
Publication Type: Abstract
Publication Acceptance Date: October 28, 2005
Publication Date: December 1, 2005
Citation: Saparno, A., Alexander, N.J., Blackwell, B., McCormick, S.P., Desjardins, A.E., Anoop, V., Kelso, S., Sprott, D., Robert, L., Tinker, N., Gleddie, S., Ouellet, T. 2005. Functional genomics and proteomics studies of F. graminearum [abstract]. The Canadian Workshop on Fusarium Head Blight. p. 70.

Technical Abstract: A 4.5K unigene F. graminearum cDNA microarray produced in-house has been used in time-course experiments to profile Fg gene expression during liquid culture conditions favouring trichothecene production (with concurrent metabolic profiling). In-depth analysis of microarray results (three biological replicates) has identified candidate genes that may play significant roles in Fg pathogenicity. The gene expression patterns of 19 genes were validated by Northern and/or Real-Time PCR analysis. Two biological replicates of these liquid cultures have been subjected to iTRAQ analysis (isotope-coded-affinity-tagged proteomics technology), identifying 330 and 944 Fg proteins, respectively, and providing quantitative protein expression data over the liquid culture time course. The microarray and iTRAQ data were compared to identify common genes/proteins. In particular, gene and protein expression of a novel Fg gene cluster are co-ordinately induced during trichothecene-producing liquid culture conditions. Disruption and add-back analysis of one gene within this gene cluster suggests it contributes to butenolide production. Preliminary results from Fg Affymetrix data indicate that this gene cluster is expressed in planta, during Fg infection of maize ears.