ELISA Analysis for Fusarium in Barley: Development of Methodology and Field Assessment

Abstract

Evaluating Fusarium head blight (FHB) involves inoculating barley (Hordeum vulgare L.) with Fusarium graminearum Schwabe [teleomorph Gibberella zeae (Schwein.) Petch] followed by visual observation of disease and analysis for deoxynivalenol (DON). Disease symptoms and DON are not always correlated because both are affected by environmental variables. The objective of this study was to develop an enzyme‐linked immunosorbent assay (ELISA) for quantification of FHB in barley. Antibodies to F. graminearum were tested for reaction with other Fusarium spp. Antibodies from cell line IF8 reacted with Fusarium spp. tested, but not other Ascomycota. The ELISA method was developed using seed lots with no, low, and high levels of DON. Quantity of seed, volume of extraction buffer, and agitation time were tested and Fusarium quantified with ELISA. Five genotypes each for high, medium, and low ELISA values were selected from a field experiment using a doubled‐haploid mapping population in 2003. The lines were grown in 2004 and scored for FHB, DON, and ELISA. ELISA had lower error than FHB or DON. Lines selected for low, medium, and high ELISA in 2003 had low, medium, and high ELISA values in 2004. ELISA and DON were correlated in both field experiments (r = 0.51). ELISA and DON were correlated (r = 0.71) in samples selected from grain elevators in 1993 through 2003 indicating naturally occurring Fusarium spp. outside the B clade had no effect on the performance of the ELISA analysis.