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Abscisic acid (ABA) has been well defined as an important stress hormone in plants. The signaling pathway of ABA involves a family of pyrabactin resistant-like-1 PYR/PYL/RCAR receptors (PYL receptors) that bind ABA and form a complex with a protein phosphatase 2C (PP2C) family member resulting in downstream signaling events. The ABA receptor family has been well characterized in the model dicot Arabidopsis thaliana and more recently this characterization has branched out into cereals Oryza sativa (rice) and Hordeum vulgare (barley), as well as the monocot model plant Brachypodium distachyon and Fragaria vesca (strawberry). The analysis of these characterized ABA receptors and the use of online databases has allowed the identification of multiple putative ABA receptors in Triticum aestivum (wheat). ABA has been historically called a positive effector. Overexpression of proteins in the ABA signalling pathway or exogenous application of ABA is known to cause an increase in drought, cold, and salt tolerance. More recently ABA has been linked to increased fungal susceptibility in several plants. The role ABA plays in the biotic stress response is still largely unexplored. The focus of this project was to identify and characterize a putative wheat ABA receptor through bioinformatics and an in vitro enzyme activity assay, and use virus induced gene silencing (VIGS) to test what role this receptor plays in fugal susceptibility. A total of 13 putative ABA receptors were located, nine of which are unique between the wheat subgenomes. One receptor TaPYL5.1 was recombinantly expressed, purified, and confirmed as an ABA receptor through a phosphatase based enzyme activity assay. A receptor with high sequence identity to TaPYL5.1, TaPYL5.2A, was targeted for plant trials because the TaPYL5.1 plasmid sequence was codon optimized. A VIGS approach was used to knock down TaPYL5.2A in planta. The TaPYL5.2A knockdown plants were found to have an increased resistance to Fusarium Head Blight progression in the early stages of the disease. In conclusion, wheat ABA receptors were successfully identified and an important correlation between decreased receptor levels and increased early Fusarium Head Blight resistance was found. This correlation however was not easily reproducible due to the severity of coupling VIGS with Fusarium Head Blight, and should be followed up with additional studies looking at the broader family of wheat ABA receptors.

Fusarium head blight (FHB) caused by Fusarium graminearum is a fungal disease of wheat that can result in severe yield losses and contaminate grain with deoxynivalenol (DON). Wheat cultivars with different levels of FHB resistance were combined with fungicides application to control FHB. Results showed that foliar fungicide Prosaro™ combined with moderately resistant cultivars greatly reduced the risk of FHB. Integrating fungicide application with moderately resistant cultivars can be an effective strategy in controlling FHB. Quantitative trait loci (QTL) for resistance to FHB related traits were analyzed using a double haploid population. Four QTL associated with FHB resistance was detected on chromosomes 2B, 2D, 4D and 7A. The QTL on chromosome 2B and 4D were found to reduce multiple FHB-related traits and were more frequently detected than QTL on chromosome 2D and 7A. QTL on chromosome 2B and 4D could be valuable for improving FHB resistance in wheat. / October 2015

Fusarium head blight (FHB) continues to threaten the economic sustainability of wheat and barley production in Canada and worldwide. The overall goal of this thesis is to expand our current knowledge of the FHB pathogen, Fusarium graminearum and its trichothecene chemotype diversity. Continuous monitoring of trichothecene chemotypes may well inform on the potential risk and the type of Fusarium populations present in a given region. Fusarium populations in Winnipeg and Carman, Manitoba were examined using chemotype as a marker in the field. Rapid expansion of the 3-acetyldeoxynivalenol (3-ADON) chemotype was observed in Winnipeg and Carman. 3-ADON chemotype is consistently found at high frequencies over the previously common 15-acetyldeoxynivalenol (15-ADON) chemotype, suggesting that the shift in pathogen populations is continuing. This study provides the first evidence on the presence of nivalenol (NIV) producing F. cerealis strains in winter wheat in Manitoba, Canada. Therefore, discovery of NIV producing F. cerealis in wheat poses a serious concern for the wheat industry in Canada. Phylogenetic, chemotypic, phenotypic, and pathogenic abilities of 150 strains of F. graminearum species complex (FGSC) from eight countries were investigated. Type and amount of trichothecenes produced by a strain are key factors in determining the level of aggressiveness of that strain regardless of its species origin. The sequence variations of TRI8 gene in different species in the FGSC were examined as Fusarium species may produce different types of trichothecenes depending on differences in the core trichothecene (TRI) cluster genes. The TRI8 haplotypes did group according to chemotype rather than by species, indicating that 3-ADON, 15-ADON and NIV chemotypes have a single evolutionary origin. Comparison of TRI gene expression demonstrated that accumulation of TRI transcripts was higher in 3-ADON producing strains compared to 15-ADON and NIV strains. The presence of masked mycotoxins deoxynivalenol-3-glucoside (D3G) in food and feed is an increasing concern. Canadian spring wheat cultivars inoculated with different chemotypes produce D3G upon Fusarium infection and moderately resistant/intermediate cultivars showed higher D3G/DON ratio compared to susceptible cultivars. / October 2016

Plant-associated microbes (collectively the microbiome) are important contributors to plant health. They are known to play roles in increasing yield via improving stress tolerance, promoting growth, and suppressing the activity of plant pathogens. We investigated the wheat seed-head microbiome (phytomicrobiome) as a potential source of resistance to Fusarium head blight (FHB), or scab. FHB is a devastating disease in wheat, and other cereal grains, that causes losses in both quantity, through reduced yield, and quality of grain, through the production of toxins such as Deoxynivalenol. Efforts to combat FHB have focused primarily on breeding cultivars with resistance and applying fungicides. However, new resources for combatting FHB may lie in microbiome-plant interactions. To explore host-microbiome-pathogen interactions, we used field trials to characterize the seed head bacterial community (16S rRNA gene amplicons) across planting locations, host resistance genotypes, varieties, and plant development stages. We identified bacterial amplicon sequence variants (ASVs) present in each sample and then examined ASV community composition based on our variables. Characterizing bacterial relative abundance across samples, we identified 9,063 ASVs. These ASVs clustered according to plant developmental stages or maturity plant, location, and host genotype, but not by variety or maturity group. First, comparing plants at the pre-flowering versus mature grain-head stage, we found that both bacterial community richness and evenness changed significantly. In addition to these developmental changes, we found that bacterial community structure changes across locations, even between locations. Finally, we found that, in the presence of the pathogen, ASVs cluster by host resistance genotype, and that there are important taxonomic groups that are differentially abundant in the presence of the pathogen. Overall, we found that the wheat grain-head microbiome is shaped by environment-host-pathogen interactions, and that these interactions lead to differential abundance of particular community members that may be important in the management of FHB. / Master of Science in Life Sciences / Plant associated microbes are important contributors to plant health. They are known to play roles in increasing yield via improved stress tolerance, promoting growth, and suppressing plant disease. We investigated the wheat grain-head microbial communities as a source of disease resistance. The disease is called Fusarium Head Blight (FHB) and is caused by Fusarium graminaerum. FHB is a devastating disease in wheat and other cereals, causing losses, through reduced yield and quality through the production of toxins that prohibit use of the grain. To combat FHB, research has focused on developing plants that have resistance and the application of chemical fungicides. However, new resources for combating FHB may lie in the interactions between plants and microbes. This research is focused on identifying microbes that naturally interact with the plant, and how the pathogen, Fusarium, interacts with these beneficial microbes. In field trials, we characterized the microbial community by DNA sequencing technologies across locations, wheat with varying levels of genetic resistance, and wheat developmental stages. First, between the wheat kernel samples of pre-flowering and maturity, we found significant differences in microbial community. Consistent with other studies we found that the largest changes in microbial community composition across different growing locations. Finally, we found an interaction between the grain head microbiome and host resistance state when plants were exposed to the pathogen. Overall, we find that the wheat grain head microbiome is shaped by growing location and through interactions with the plant host and pathogen.

Fusarium head blight (FHB) caused by Fusarium graminearum and Septoria tritici blotch (STB) caused by Septoria tritici are economically important wheat diseases in Ontario. Both reduce yield, FHB is associated with mycotoxin accumulation including deoxynivalenol (DON). Different F. graminearum chemotypes produce either DON/15-acetyldeoxynvialenol (ADON) or DON/3-ADON. The majority (97.5%) of F. graminearum isolates collected from commercial fields across Ontario were 15-ADON chemotype, the remaining were 3-ADON. In inoculated field experiments 3-ADON chemotypes were more aggressive and toxic compared with 15-ADON chemotypes as measured by FHB symptoms and DON content. In inoculated field experiments with a population derived from ‘RCATL33’ and ‘RC Strategy’ soft red winter wheat parents, genetic resistance was more effective than fungicide application at controlling FHB. Field trials with the hard red winter wheat population derived from ‘Maxine’ and ‘FTHP Redeemer’ parents revealed that STB and FHB phenotypic resistance had no negative impact on grain yield in the absence of disease.

Master of Science / Agronomy / Allan Fritz / Fusarium head blight (FHB) is a fungal disease, mostly commonly associated with F. graminearum, which affects cereal crops such as wheat resulting in substantial yield losses and reductions in grain quality. The onset of the disease can occur rapidly when warm, wet or humid weather coincides with flowering in the spring. The pathogen also produces mycotoxins such as deoxynivalenol (DON) that accumulate in the grain and can be toxic to humans and animals. This results in additional economic losses as contaminated grain must be discarded or blended to reduce the amount of toxin in order to meet federal regulatory limits. Development and deployment of resistant cultivars has proved to be an effective method to combat the disease, and many resistant sources have been reported in the literature with the majority of major resistance coming from Chinese landraces. Transferring resistance from these sources into cultivars adapted to the U.S. has been a slow process due to linkage of FHB resistance genes with poor agronomic traits. Therefore, it is important for breeders to search for sources of resistance in native material adapted to their local conditions. In this study, we aimed to identify quantitative trait loci (QTL) for resistance to spread of FHB within the head (Type II resistance), accumulation of DON toxin in grain (Type III resistance), and resistance to kernel infection (Type IV resistance). Plant material consisted of 148 doubled haploid (DH) lines from a cross between the two moderately resistant hard red winter wheat (HRWW) cultivars Art and Everest. The study was conducted for two years using a point inoculation technique in a greenhouse in Manhattan, KS. Three QTL conferring resistance to FHB traits were detected on chromosomes 2D, 4B, and 4D. The QTL on chromosomes 4B and 4D overlapped with the major height genes Rht1 and Rht2, respectively. Plant height has shown previous associations with FHB, though the underlying cause of these associations is not well understood. The majority of results have reported increased susceptibility associated with shorter plant types; however, in this study, the haplotype analysis for the Rht-B1 and Rht-D1 loci showed an association between the dwarfing alleles and increased resistance to FHB. This suggests either pleiotropic effects of these loci or perhaps linkage with nearby genes for FHB resistance. Markers close to the peaks of the FHB resistance QTL have the potential for Kompetitive Allele Specific PCR (KASP) marker development and subsequent use in marker assisted selection (MAS) to help improve overall FHB resistance within breeding programs.

Challenges to growing winter wheat in North Dakota include winter temperature and disease pressure. Fusarium head blight (FHB) is a devastating disease that necessitates breeding for resistance. In the NDSU breeding program FHB resistance genes are often associated with a decrease in performance. This study used single seed descent to advance lines while maintaining a near random population. Early generation (F4) selection focused heavily on yield and the presence of FHB resistance quantitative trait loci to develop winter wheat lines with FHB resistance and high yield. Stripe rust is a fungal disease that is becoming increasingly problematic in North Dakota. To assess the available stripe rust resistance in the NDSU winter wheat germplasm, two sets of diverse breeding lines were used for stripe rust resistance phenotyping and genotyping by sequencing. The phenotype and genotype data were then used to locate resistance genes through genome wide association study.

The global food supply is being stressed by climate change, a growing population, and harmful diseases. One risk to vital cereal crops such as wheat and barley is Fusarium head blight (FHB), caused by the fungal plant pathogen Fusarium graminearum. Ascospores of the fungus are released from perithecia on the residues of corn and small grains and can be transported long distances (>500 m) through the atmosphere. The overall objective of this work was to assess the influence of meteorological conditions and perithecial maturity on ascospore release. The research focuses on F. graminearum because of its damaging impact to staple crops and the global ubiquity of FHB. The first specific objective was to apply state-of-the-science techniques to identify causal meteorological variables of ascospore release. We analyzed field measurements of airborne ascospores against meteorological conditions at Virginia Tech's Kentland Farm, Blacksburg, Virginia, USA and used convergent cross mapping and multivariate state space reconstruction to identify significant causal agents within this complicated natural and dynamic system. We identified relative humidity, solar radiation, wind speed, and air temperature as predictors of ascospore release. Our second research objective was to understand the impact of varying meteorological conditions on ascospore release under controlled environmental conditions. We assessed ascospore release in a chamber with controlled temperature (15°C and 25°C) and relative humidity (60%, 75%, and 95%). Ascospores released from ascospore-producing structures (perithecia) were captured on microscope slides placed inside of 3D-printed ascospore discharge devices. Results showed the sensitivity of ascospore release to relative humidity and temperature, with cool temperature and high relative humidity resulting in greater quantities of ascospores released. Our third research objective was to determine the relationship between the maturity, the number of ascospores, and the hardness of perithecia. A mechanical compression testing instrument was used to investigate the hardness of perithecia at various stages of maturity, producing a mean perithecium compression constant quantifying the uniaxial compression force required to rupture a perithecium. Results indicated that old perithecia contain the greatest amount of ascospores and exhibit increased resiliency, requiring greater forces to rupture, compared to young perithecia. This research has illustrated the complexities of F. graminearum ascospore release by describing the impact of several meteorological conditions and perithecial maturity on the timing and quantity of released ascospores. Collectively, our results may inform wheat growers on the nature and timing of ascospore release, which could help inform FHB management decisions in the future. / Ph. D.

No description available.