Biosynthesis of deoxynivalenol in spikelets of barley inoculated with macroconidia of Fusarium graminearum

This research examined the biosynthesis of deoxynivalenol (DON) and 15-acetyldeoxynivalenol (15-ADON) in barley spikelets inoculated with macroconidia of Fusarium graminearum (Group-II). Investigations were conducted to determine if these toxins were present in macroconidia of the pathogen prior to inoculating barley spikelets. Extracts of macroconidia cultured from mung bean agar were analyzed using gas chromatography-mass spectrometry. Neither DON or 15-ADON was detected in three isolates' macroconidia when compared with macroconidia-DON-matrix standards adjusted to 100, 200, 300, and 400 ng/g with a detection limit of 100 ng/g. Mean recovery of DON that was added to macroconidia was 89.5%. The same isolates were pathogenic on barley cultivars Robust (moderately susceptible) and Chevron (moderately resistant) and produced DON (0 to 3.69 ng/g) and 15-ADON (detected but not quantified) when grown in rice culture. Greenhouse experiments were performed to determine when DON and 15-ADON were detectable after inoculation and to quantify their amount in inoculated barley spikelets. The three isolates of F. graminearum were separately inoculated to a central spikelet on heads of barley cultivars Robust and Chevron. Both toxins were detected in spikelets 48 h postinoculation (PI). DON increased dramatically after 72 h and did not diminish thereafter. Accumulation of 15-ADON peaked at 72 to 120 h and decreased by 240 h PI. There were no statistical differences between cultivars or among fungal isolates for accumulation of either toxin when averaged over the time intervals. Differences of toxin accumulation at each sampling interval were significant (P < 0.0001) when averaged over isolates and cultivars. Spikelets of six cultivars and lines were sampled at inoculation and 18, 36, 54, 72, and 90 h PI. DON and 15-ADON were detected at 36 h PI, but differences among the cultivars and lines were not significant. Yield of DON in inoculated spikelets of 31 barley cultivars and lines at 72 h PI ranged from 0.14 to 1.26 mu g per spikelet, and differences among the cultivars and lines were significant (P < 0.002). The data demonstrate a useful range of variability for toxin accumulation in inoculated spikelets among germ plasm in the Minnesota breeding program. Macroconidia with no detectable DON or 15-ADON could be used for in vitro studies of toxin biosynthesis. Establishing when DON and 15-ADON are synthesized facilitates studying the effects of promising fungicides, biocontrol organisms, and new or novel genetic resistance mechanisms and if or how they may prevent or delay the biosynthesis of toxins.