A bioassay for predicting the resistance of wheat leaves to Septoria nodorum

Plants of the winter wheat cv. Bounty were inoculated with Septoria nodorum and then assayed for ergosterol. A detached leaf technique was used in which leaf segments were incubated on agar containing benzimidazole. Four levels of a conidial suspension inoculum were applied using a measured droplet technique. Ergosterol in plant extracts was measured using HPLC and its identity confirmed by co-injection with pure ergosterol as an internal standard. The extracts were also assayed by silica-gel thin layer chromatography with the sterol being visualised with rhodamine; the presence of ergosterol in the material from the two higher disease level treatments was confirmed by analysing the eluted spots in a spectrophotometer. Additional confirmation of ergosterol was obtained using gas-liquid chromatography comparing a mixture of known plant sterols with a sample from the diseased leaf tissue. The ergosterol assay was found to be very sensitive and offers a high level of reproducibility. It would therefore appear that such assays could be of value in breeding programmes when it is necessary to screen wheat cultivars for their reaction to S. nodorum or other fungal pathogens.