ARS | Publication request: DEVELOPMENT OF A QUANTITATIVE PCR ASSAY TO DETERMINE FUSARIUM GRAMINEARUM LOAD IN CONTAMINATED WHEAT SEEDS

Submitted to: Nucleic Acids Research
Publication Type: Abstract Only
Publication Acceptance Date: June 23, 2004
Publication Date: June 23, 2004
Citation: Dyer, R.B., Kendra, D.F., Brown, D.W. 2004. Development of a quantitative pcr assay to determine fusarium graminearum load in contaminated wheat seeds. Nucleic Acids Research.

Technical Abstract: Fusarium graminearum, the predominant causal agent of wheat head scab, is a significant food safety concern and has a major economic impact on agriculture. In addition to a reduction in yield, infected wheat may also lead to seed contamination by the fungal generated toxin, deoxynivalenol (DON). DON is toxic to both plants and animals and is also a critical component of pathogenicity of wheat. Wheat contaminated with DON exacerbates the economic loss during grain marketing through added expenses for screening and testing. Given these concerns, it is imperative that we understand the pathway(s) for DON synthesis so that effective control strategies can be developed. Through gene knockout technology, genes and pathways suspected of being important in the pathogenesis of F. graminearum are being tested in the greenhouse and in the field. These studies depend on technology that allows the researcher to distinguish between disease caused by the parental strain and disease caused by the mutant, test strain. The recent advance of quantitative PCR has enabled the detection and quantification of fungi from field samples at the genus and species levels. Nicholson, et al. (Physiological and Molecular Plant Pathology, 53:17-37) has developed primers (Fg16F/R) to specifically detect and quantify F. graminearum from infected grain. In this report, we describe the development of a real-time quantitative PCR protocol using the Fg16F/R primers. In addition, we developed primers specific to our gene knockout technology. Given the documented specificity of the Fg16F/R primers and the widespread use of gene knockout technology, our protocol will be of general use to the plant pathology community for studying the pathobiology of F. graminearum.