Resistance to Fusarium Head Blight and Deoxynivalenol Accumulation in Virginia Barley.

Fusarium graminearum (teleomorph Gibberella zeae), is a devastating disease of barley (Hordeum vulgare) in the United States. Recent epidemics of FHB in the mid-Atlantic region have underscored the need to develop new commercial varieties of barley that are resistant to FHB and restrict accumulation of the mycotoxin deoxynivalenol (DON). FHB incidence, FHB index, and DON levels of Virginia hulled and hulless barley genotypes were evaluated over five years (2006 to 2010) in FHB nurseries in Virginia. FHB incidence ranged from 22.5% (2010) to 80.1% (2008), and mean DON levels ranged from 0.5 ± 0.4 (2008) to 2.4 ± 2.1 ppm (2010). Barley genotype played a significant role in determining FHB resistance in 2006 to 2009. DON levels were significantly different among barley genotypes in 2007, 2008, and 2009. FHB incidence was positively correlated with FHB index in all 5 years studied. In 2006 and 2010, FHB incidence and index were positively correlated with DON. Early spike emergence resulted in higher FHB incidence and index in 2007, 2008, and 2010. This preliminary work has identified some promising hulled and hulless barley genotypes for targeted breeding and commercialization efforts in FHB nurseries in the future; 'Eve' (hulless) and 'Thoroughbred' (hulled) ranked among the most FHB resistant genotypes.

A comparison of two milling strategies to reduce the mycotoxin deoxynivalenol in barley.

Winter barley (Hordeum vulgare L.), a potential feedstock for fuel ethanol production, may be contaminated with the trichothecene mycotoxin deoxynivalenol (DON). DON is a threat to feed and food safety in the United States and may become concentrated during the production of distillers dried grains with solubles (DDGS). DDGS is a coproduct of fuel ethanol production and is increasingly being used as feed for domestic animals. Therefore, new strategies to reduce the threat of DON in DDGS need to be developed and implemented for grain destined for fuel ethanol production. It is known that large concentrations of DON accumulate in the hulls of wheat and barley. Consequently, improved methods are needed to carefully remove the hull from the grain and preserve the starchy endosperm. Whole kernels from five Virginia winter barley genotypes were used to evaluate the abilities of two different milling strategies (roller milling and precision milling (FitzMill)) for their ability to remove the hull-enriched tissue from the kernel while maintaining starch levels and reducing DON levels in the endosperm-enriched tissue. After whole kernels were milled, DON and starch levels were quantified in the hull-enriched fractions and endosperm-enriched fractions. Initial milling experiments demonstrated that the precision mill system (6 min run time) is able to reduce more DON than the roller mill but with higher starch losses. The average percent DON removed from the kernel with the roller mill was 36.7% ± 5.5 and the average percent DON removed from the dehulled kernel with the precision mill was 85.1% ± 9.0. Endosperm-enriched fractions collected from the roller mill and precision mill contained starch levels ranging from 49.0% ± 12.1 to 59.1% ± 0.5 and 58.5% ± 1.6 to 65.3% ± 3.9, respectively. On average, the precision mill removed a mass of 23.1% ± 6.8 and resulted in starch losses of 9.6% ± 6.3, but produced an endosperm-enriched fraction with relatively very little average DON (5.5 ± 2.7 µg g(-1)). In contrast, on average, the roller mill removed a mass of 12.2% ± 1.6 and resulted in starch losses of 2.1% ± 0.5, but produced an endosperm-enriched fraction with high average DON (20.7 ± 13.5 µg g(-1)). In a time course precision milling experiment, we tested barley genotypes Nomini, Atlantic, and VA96-44-304 and attempted to reduce the starch loss seen in the first experiment while maintaining low DON concentrations. Decreasing the run time of the precision mill from 5 to 2 min, reduced starch loss at the expense of higher DON concentrations. Aspirated fractions revealed that the precision milled hull-enriched fraction contained endosperm-enriched components that were highly contaminated with DON. This work has important implications for the reduction of mycotoxins such as DON in barley fuel ethanol coproducts and barley enriched animal feeds and human foods.

Conversion of deoxynivalenol to 3-acetyldeoxynivalenol in barley-derived fuel ethanol co-products with yeast expressing trichothecene 3-O-acetyltransferases.

BACKGROUND: The trichothecene mycotoxin deoxynivalenol (DON) may be concentrated in distillers dried grains with solubles (DDGS; a co-product of fuel ethanol fermentation) when grain containing DON is used to produce fuel ethanol. Even low levels of DON (≤ 5 ppm) in DDGS sold as feed pose a significant threat to the health of monogastric animals. New and improved strategies to reduce DON in DDGS need to be developed and implemented to address this problem. Enzymes known as trichothecene 3-O-acetyltransferases convert DON to 3-acetyldeoxynivalenol (3ADON), and may reduce its toxicity in plants and animals. RESULTS: Two Fusarium trichothecene 3-O-acetyltransferases (FgTRI101 and FfTRI201) were cloned and expressed in yeast (Saccharomyces cerevisiae) during a series of small-scale ethanol fermentations using barley (Hordeum vulgare). DON was concentrated 1.6 to 8.2 times in DDGS compared with the starting ground grain. During the fermentation process, FgTRI101 converted 9.2% to 55.3% of the DON to 3ADON, resulting in DDGS with reductions in DON and increases in 3ADON in the Virginia winter barley cultivars Eve, Thoroughbred and Price, and the experimental line VA06H-25. Analysis of barley mashes prepared from the barley line VA04B-125 showed that yeast expressing FfTRI201 were more effective at acetylating DON than those expressing FgTRI101; DON conversion for FfTRI201 ranged from 26.1% to 28.3%, whereas DON conversion for FgTRI101 ranged from 18.3% to 21.8% in VA04B-125 mashes. Ethanol yields were highest with the industrial yeast strain Ethanol Red®, which also consumed galactose when present in the mash. CONCLUSIONS: This study demonstrates the potential of using yeast expressing a trichothecene 3-O-acetyltransferase to modify DON during commercial fuel ethanol fermentation.