The fungal plant pathogen Fusarium graminearum Schwabe (teleomorph Gibberella zeae¬) produces a dangerous trichothecene mycotoxin called deoxynivalenol (DON) and causes a devastating disease of barley (Hordeum vulgare L.) called Fusarium head blight (FHB). Food and feed products derived from barley, such as dried distillers grains with solubles (DDGS), may be contaminated with DON and pose a threat to the health of humans and domestic animals. New methods to mitigate the threat of DON in barley need to be developed and implemented. TRI101 and TRI201 are trichothecene 3-O-acetyltransferases that modify DON and reduce its toxicity. The first objective of this research was to isolate unique TRI101 and TRI201 enzymes that modify DON efficiently. We hypothesized that TRI101/TRI201 enzymes from different species of Fusarium would have varying rates and abilities to modify DON. Using degenerate primers, an internal portion of TRI101 or TRI201 was identified in 54 strains of Fusarium. Full-length sequences of seven TRI101 or TRI201 genes were cloned and expressed in yeast. All seven genes acetylated DON, but at different rates. The second objective of this research was to utilize transformed yeast expressing TRI101 or TRI201 to reduce DON levels in barley mashes and ultimately in DDGS. We hypothesized that DON levels would be reduced in DDGS derived from mashes prepared with transformed yeast. Five different barley genotypes were used to prepare the fermentation mashes and DON levels were reduced in all DDGS samples derived from mashes prepared with transformed yeast. The third objective of this study was to characterize barley genotypes developed at Virginia Tech for resistance to FHB and DON. We hypothesized that significant differences in resistance would be observed among barley genotypes and FHB resistance would be associated with reduced DON accumulation. From 2006 to 2010, FHB resistance was assessed in hulled (22 to 37) and hulless (13 to 32) barley genotypes by measuring incidence and index, and DON resistance was determined by quantifying DON levels in ground grain using gas chromatography-mass spectrometry. Our study showed that FHB and DON resistance is significantly determined by genotype. The final objective of this study was to develop a robust tissue culture system necessary for future development of transformed barley plants with FHB resistance gene(s). We hypothesized that callus production would vary among barley genotypes. In our analysis of 47 Virginia barley genotypes, 76% (36/47) of the genotypes produced callus tissue and there were significant differences in callus size. Our work sets the stage for identifying and characterizing DON detoxification genes in the future. The development of commercial barley lines that do not accumulate DON and that are resistant to FHB will directly impact growers and producers of small grains in the eastern U.S. / Ph. D.

Fusarium head blight (FHB), caused mainly by Fusarium graminearum (teleomorph: Gibberella zeae), is one of the most damaging diseases of wheat. A ‘Brio’/‘TC 67’ spring wheat population was used to map quantitative trait loci (QTLs) for resistance to FHB, and to study the association of morphological and developmental characteristics with FHB resistance. Interval mapping (IM) detected a major QTL on chromosome 5AL for resistance to disease severity (type II resistance) and Fusarium-damaged kernels (FDK) under greenhouse and field conditions, respectively. Inconsistent QTL(s) was also detected on chromosome 5BS for disease severity and index using field data. The associations of plant height and number of days to anthesis were negative with disease incidence, severity, index, and deoxynivalenol (DON) accumulation data under field conditions. However, number of days to anthesis was positively correlated with disease severity (greenhouse) and FDK (field). Awnedness had a negative effect on FHB, namely the presence of awns resulted in less disease in the population. Spike threshability also affected FHB so that the hard threshable genotypes represented lower disease. Phylogenetic relationships of putative F. graminearum isolates from different sources were characterized using Tri101 gene sequencing data. Canadian and Iranian isolates clustered in F. graminearum lineage 7 (=F. graminearum sensu stricto) within the F. graminearum clade while the isolates received from CIMMYT, Mexico were placed in F. graminearum lineage 3 (=Fusarium boothii) within the Fg clade or Fusarium cerealis. The PCR assay based on the Tri12 gene revealed the presence of the NIV, 3-ADON, and 15-ADON chemotypes with 15-ADON being the predominant chemotype. While we did not find the NIV chemotype among the Canadian isolates, it was the predominant chemotype among the Iranian isolates. High variation in aggressiveness was observed among and within Fusarium species tested, with the isolates of F. graminearum sensu stricto being the most aggressive and the NIV chemotype being the least aggressive. The interactions between Fusarium isolates and wheat genotypes from different sources were investigated by inoculating isolates of F. graminearum sensu stricto and F. boothii on wheat genotypes. Significant differences were observed among the genotypes inoculated by single isolates. Results also showed significant interactions between Fusarium isolates and wheat genotypes. The F. boothii isolates from CIMMYT produced low disease symptom and infection on wheat genotypes regardless of the origin of the genotypes while F. graminearum sensu stricto isolates from Canada and Iran resulted in higher FHB scores.

Fusarium head blight (FHB), caused mainly by Fusarium graminearum (teleomorph: Gibberella zeae), is one of the most damaging diseases of wheat. A ‘Brio’/‘TC 67’ spring wheat population was used to map quantitative trait loci (QTLs) for resistance to FHB, and to study the association of morphological and developmental characteristics with FHB resistance. Interval mapping (IM) detected a major QTL on chromosome 5AL for resistance to disease severity (type II resistance) and Fusarium-damaged kernels (FDK) under greenhouse and field conditions, respectively. Inconsistent QTL(s) was also detected on chromosome 5BS for disease severity and index using field data. The associations of plant height and number of days to anthesis were negative with disease incidence, severity, index, and deoxynivalenol (DON) accumulation data under field conditions. However, number of days to anthesis was positively correlated with disease severity (greenhouse) and FDK (field). Awnedness had a negative effect on FHB, namely the presence of awns resulted in less disease in the population. Spike threshability also affected FHB so that the hard threshable genotypes represented lower disease. Phylogenetic relationships of putative F. graminearum isolates from different sources were characterized using Tri101 gene sequencing data. Canadian and Iranian isolates clustered in F. graminearum lineage 7 (=F. graminearum sensu stricto) within the F. graminearum clade while the isolates received from CIMMYT, Mexico were placed in F. graminearum lineage 3 (=Fusarium boothii) within the Fg clade or Fusarium cerealis. The PCR assay based on the Tri12 gene revealed the presence of the NIV, 3-ADON, and 15-ADON chemotypes with 15-ADON being the predominant chemotype. While we did not find the NIV chemotype among the Canadian isolates, it was the predominant chemotype among the Iranian isolates. High variation in aggressiveness was observed among and within Fusarium species tested, with the isolates of F. graminearum sensu stricto being the most aggressive and the NIV chemotype being the least aggressive. The interactions between Fusarium isolates and wheat genotypes from different sources were investigated by inoculating isolates of F. graminearum sensu stricto and F. boothii on wheat genotypes. Significant differences were observed among the genotypes inoculated by single isolates. Results also showed significant interactions between Fusarium isolates and wheat genotypes. The F. boothii isolates from CIMMYT produced low disease symptom and infection on wheat genotypes regardless of the origin of the genotypes while F. graminearum sensu stricto isolates from Canada and Iran resulted in higher FHB scores.