Fusarium Head Blight of Small Grains in Pennsylvania: Unravelling Species Diversity, Toxin Types, Growth, and Triazole Sensitivity.

Fusarium graminearum is the main causal species of Fusarium head blight (FHB) globally. Recent changes in the trichothecene (toxin) types in the North American FHB pathogens support the need for continued surveillance. In this study, 461 isolates were obtained from symptomatic spikes of wheat, spelt, barley, and rye crops during 2018 and 2019. These were all identified to species and toxin types using molecular-based approaches. An additional set of 77 F. graminearum isolates obtained from overwintering crop residues during winter 2012 were molecularly identified to toxin types. A subset of 31 F. graminearum isolates (15 15-acetyl-deoxynivalenol [15ADON] and 16 3-acetyl-deoxynivalenol [3ADON]) were assessed for mycelial growth, macroconidia, perithecia, and ascospore production, and sensitivity to two triazoles. Ninety percent of isolates obtained from the symptomatic spikes (n = 418) belonged to F. graminearum, with four other species found at a lower frequency (n = 39). The F. graminearum isolates from symptomatic spikes were mainly of the 15ADON (95%), followed by 3ADON (4%), nivalenol (0.7%), and NX-2 (0.3%) toxin types. All F. graminearum isolates obtained from overwintering residue were of the 15ADON type. The toxin types could not be differentiated based on the multivariate analysis of growth and reproduction traits. All isolates were sensitive to tebuconazole and metconazole fungicides in vitro. This study confirms the dominance of F. graminearum and suggests ecological and environmental factors, to be further identified, that lead to similar composition of toxin types in the northern United States. Our results may be useful to assess the sustainability of FHB management practices and provide a baseline for future FHB surveys.

Population Genetic Structure and Chemotype Diversity of Fusarium graminearum Populations from Wheat in Canada and North Eastern United States.

Fusarium head blight (FHB) is a major disease in wheat causing severe economic losses globally by reducing yield and contaminating grain with mycotoxins. In Canada, Fusarium graminearum is the principal etiological agent of FHB in wheat, producing mainly the trichothecene mycotoxin, deoxynivalenol (DON) and its acetyl derivatives (15-acetyl deoxynivalenol (15ADON) and 3-acetyl deoxynivalenol (3ADON)). Understanding the population biology of F. graminearum such as the genetic variability, as well as mycotoxin chemotype diversity among isolates is important in developing sustainable disease management tools. In this study, 570 F. graminearum isolates collected from commercial wheat crops in five geographic regions in three provinces in Canada in 2018 and 2019 were analyzed for population diversity and structure using 10 variable number of tandem repeats (VNTR) markers. A subset of isolates collected from the north-eastern United States was also included for comparative analysis. About 75% of the isolates collected in the Canadian provinces of Saskatchewan and Manitoba were 3ADON indicating a 6-fold increase in Saskatchewan and a 2.5-fold increase in Manitoba within the past 15 years. All isolates from Ontario and those collected from the United States were 15ADON and isolates had a similar population structure. There was high gene diversity (H = 0.803-0.893) in the F. graminearum populations in all regions. Gene flow was high between Saskatchewan and Manitoba (Nm = 4.971-21.750), indicating no genetic differentiation between these regions. In contrast, less gene flow was observed among the western provinces and Ontario (Nm = 3.829-9.756) and USA isolates ((Nm = 2.803-6.150). However, Bayesian clustering model analyses of trichothecene chemotype subpopulations divided the populations into two clusters, which was correlated with trichothecene types. Additionally, population cluster analysis revealed there was more admixture of isolates among isolates of the 3ADON chemotypes than among the 15ADON chemotype, an observation that could play a role in the increased virulence of F. graminearum. Understanding the population genetic structure and mycotoxin chemotype variations of the pathogen will assist in developing FHB resistant wheat cultivars and in mycotoxin risk assessment in Canada.

Root cortical anatomy is associated with differential pathogenic and symbiotic fungal colonization in maize.

Root anatomical phenotypes vary among maize (Zea mays) cultivars and may have adaptive value by modifying the metabolic cost of soil exploration. However, the microbial trade-offs of these phenotypes are unknown. We hypothesized that nodal roots of maize with contrasting cortical anatomy have different patterns of mutualistic and pathogenic fungal colonization. Arbuscular mycorrhizal colonization in the field and mesocosms, root rots in the field, and Fusarium verticillioides colonization in mesocosms were evaluated in maize genotypes with contrasting root cortical anatomy. Increased aerenchyma and decreased living cortical area were associated with decreased mycorrhizal colonization in mesocosm and field experiments with inbred genotypes. In contrast, mycorrhizal colonization of hybrids increased with larger aerenchyma lacunae; this increase coincided with larger root diameters of hybrid roots. F. verticillioides colonization was inversely correlated with living cortical area in mesocosm-grown inbreds, and no relation was found between root rots and living cortical area or aerenchyma in field-grown hybrids. Root rots were positively correlated with cortical cell file number and inversely correlated with cortical cell size. Mycorrhizae and root rots were inversely correlated in field-grown hybrids. We conclude that root anatomy is associated with differential effects on pathogens and mycorrhizal colonization of nodal roots in maize.

FvSO regulates vegetative hyphal fusion, asexual growth, fumonisin B1 production, and virulence in Fusarium verticillioides.

Hyphal anastomosis is a hallmark of filamentous fungi and plays vital roles including cellular homoeostasis, interhyphal communication and nutrient translocation. Here we identify a gene, FvSO, in Fusarium verticillioides, a filamentous ascomycete causing maize ear and stalk rot and producing fumonisin mycotoxins. FvSO, like its Neurospora crassa homologue SO, is required for vegetative hyphal fusion. It is also essential for normal vegetative growth, sporulation, and pathogenesis. FvSO encodes a predicted WW domain protein and shares 70 % protein sequence identity with N. crassa SO. FvSO deletion mutants (ΔFvSO) had abnormal distribution of conidia size, and conidia of ΔFvSO germinated much later and slower than wild type. ΔFvSO was deficient in hyphal anastomosis, had slower radial growth and produced less fungal biomass than wild type. ΔFvSO were unable to perform anastomosis, a key feature of filamentous fungi. Interestingly, production of fumonisin B1 by ΔFvSO was significantly reduced compared to wild type. Additionally, ΔFvSO was nonpathogenic to corn ears, stalks and seedlings, likely due to defective growth and development. In conclusion, FvSO is essential for vegetative hyphal fusion and is required for normal vegetative growth and sporulation, normal levels of fumonisin production and pathogenicity in F. verticillioides. The pleiotropic nature of ΔFvSO phenotypes suggests that FvSO is likely involved in certain signalling pathways that regulate multiple cellular functions.

Methods of instruction of the incident command system and related topics at US veterinary schools.

OBJECTIVE: The Incident Command System (ICS) is an adaptable construct designed to streamline response efforts to a disaster or other incident. We aimed to examine the methods used to teach the ICS at US veterinary schools and to explore alternative and novel methods for instruction of this material. METHODS: A total of 29 US accredited veterinary schools (as of February 2012) were surveyed, and 18 of the 29 schools responded. RESULTS: The ICS and related topics were taught by both classroom methods and online instruction by most of the surveyed schools. Several of the schools used readily available Federal Emergency Management Agency and US Department of Agriculture resources to aid in instruction. Most schools used one course to teach the ICS, and some schools also used unique methods such as field exercises, drills, side-by-side training with disaster response teams, elective courses, extracurricular clubs, and externships to reinforce the ICS and related topics. Some of the surveyed institutions also utilized fourth-year clinical rotations and field deployments during actual disasters as a component of their ICS and emergency response curriculum. CONCLUSION: The ICS is being taught at some form at a significant number of US veterinary schools. Additional research is needed to evaluate the efficacy of the teaching methods of the ICS in US veterinary schools.

Induction of salicylic acid-mediated defense response in perennial ryegrass against infection by Magnaporthe oryzae.

Incorporation of plant defense activators is an innovative approach to development of an integrated strategy for the management of turfgrass diseases. The effects of salicylic acid (SA), benzothiadiazole (BTH, chemical analog of SA), jasmonic acid (JA), and ethephon (ET, an ethylene-releasing compound) on development of gray leaf spot in perennial ryegrass (Lolium perenne L.) caused by Magnaporthe oryzae were evaluated. Gray leaf spot disease incidence and severity were significantly decreased when plants were treated prior to inoculation with SA, BTH, and partially by ET but not by JA. Accumulation of endogenous SA and elevated expression of pathogenesis-related (PR)-1, PR-3.1, and PR-5 genes were associated with inoculation of plants by M. oryzae. Treatment of plants with SA enhanced expression levels of PR-3.1 and PR-5 but did not affect the PR-1 level, whereas BTH treatment enhanced relative expression levels of all three PR genes. Microscopic observations of leaves inoculated with M. oryzae revealed higher frequencies of callose deposition at the penetration sites in SA- and BTH-treated plants compared with the control plants (treated with water). These results suggest that early and higher induction of these genes by systemic resistance inducers may provide perennial ryegrass with a substantial advantage to defend against infection by M. oryzae.

One fungus, one name: defining the genus Fusarium in a scientifically robust way that preserves longstanding use.

In this letter, we advocate recognizing the genus Fusarium as the sole name for a group that includes virtually all Fusarium species of importance in plant pathology, mycotoxicology, medicine, and basic research. This phylogenetically guided circumscription will free scientists from any obligation to use other genus names, including teleomorphs, for species nested within this clade, and preserve the application of the name Fusarium in the way it has been used for almost a century. Due to recent changes in the International Code of Nomenclature for algae, fungi, and plants, this is an urgent matter that requires community attention. The alternative is to break the longstanding concept of Fusarium into nine or more genera, and remove important taxa such as those in the F. solani species complex from the genus, a move we believe is unnecessary. Here we present taxonomic and nomenclatural proposals that will preserve established research connections and facilitate communication within and between research communities, and at the same time support strong scientific principles and good taxonomic practice.

Hidden host plant associations of soilborne fungal pathogens: an ecological perspective.

Much of the current knowledge on population biology and ecology of soilborne fungal pathogens has been derived from research based on populations recovered from plants displaying disease symptoms or soil associated with symptomatic plants. Many soilborne fungal pathogens are known to cause disease on a large number of crop plants, including a variety of important agronomical, horticultural, ornamental, and forest plants species. For instance, the fungus Verticillium dahliae causes disease on >400 host plants. From a phytopathological perspective, plants on which disease symptoms have not been yet observed are considered to be nonhosts for V. dahliae. This term may be misleading because it does not provide information regarding the nature of the plant-fungus association; that is, a nonhost plant may harbor the fungus as an endophyte. Yet, there are numerous instances in the literature where V. dahliae has been isolated from asymptomatic plants; thus, these plants should be considered hosts. In this article, we synthesize scattered research that indicates that V. dahliae, aside from being a successful and significant vascular plant pathogen, may have a cryptic biology on numerous asymptomatic plants as an endophyte. Thus, we suggest here that these endophytic associations among V. dahliae and asymptomatic plants are not unusual relationships in nature. We propose to embrace the broader ecology of many fungi by differentiating between "symptomatic hosts" as those plants in which the infection and colonization by a fungus results in disease, and "asymptomatic hosts" as those plants that harbor the fungus endophytically and are different than true nonhosts that should be used for plant species that do not interact with the given fungus. In fact, if we broaden our definition of "host plant" to include asymptomatic plants that harbor the fungus as an endophyte, it is likely that the host ranges for some soilborne fungal pathogens are much larger than previously envisioned. By ignoring the potential for soilborne fungal pathogens to display endophytic relationships, we leave gaps in our knowledge about the population biology and ecology, persistence, and spread of these fungi in agroecosystems.

Deoxynivalenol biosynthesis-related gene expression during wheat kernel colonization by Fusarium graminearum.

Deoxynivalenol (DON) is a potent mycotoxin and virulence factor produced by Fusarium graminearum. We examined the expression of the core DON biosynthetic gene Tri5 during wheat head infection of susceptible and resistant cultivars and susceptible cultivars treated with strobilurin fungicides (e.g., azoxystrobin). DON was quantified to correlate expression with toxin accumulation. The highest Tri5 expression relative to housekeeping genes occurred at the infection front. As infection progressed, earliest-infected kernels showed diminished relative Tri5 expression but Tri5 expression never ceased during the 21 days observed. Azoxystrobin treatment showed no significant effect on either relative Tri5 expression or DON quantity. The resistant cultivar 'Alsen' showed minimal spread of the fungus, with no fungus detected by day 21. DON was not detected in significant quantities in Alsen in the later stages sampled. In Wheaten, DON levels were negligible at 8 days postinoculation (dpi), with detectable DON at later-sampled time points. Tri5 was detected even in fully senesced kernels 21 dpi. Our data demonstrate the presence of Tri5 transcripts in a susceptible cultivar over a much longer time period than has been previously documented. This suggests the ability of the fungus to rapidly resume toxin biosynthesis in dried infected grain should conducive environmental conditions be present, and provides a possible mechanism for high DON levels in asymptomatic grain.

Relationship of sphinganine analog mycotoxin contamination in maize silage to seasonal weather conditions and to agronomic and ensiling practices.

ABSTRACT Sphinganine analog mycotoxins (SAMs) are reported in maize and maize based feeds. Our objectives were to detect and quantify fumonisins B(1) and B(2) and Alternaria toxins (AAL toxins) AAL-TA and AAL-TB and determine how agronomic practices, weather conditions, and ensiling affected the occurrence and levels in maize silage. Silage was collected at harvest and after ensiling in 2001 and 2002 from 30 to 40 dairies, representing four regions in Pennsylvania. SAMs were quantified using high pressure liquid chromatography (HPLC) with fluorescence detection and high pressure liquid chromatography-mass spectrometry HPLC-MS. The average concentrations and ranges were as follows: fumonisin B(1) 2.02 mug/g (0.20 to 10.10), fumonisin B(2) 0.98 mug/g (0.20 to 20.30), AAL-TA 0.17 mug/g (0.20 to 2.01), and AAL-TB 0.05 mug/g (0.03 to 0.90). Fumonisin B(1) was the most frequently detected toxin (92%) in all samples, followed by fumonisin B(2) (55%), AAL-TA (23%), and -TB (13%). Temperature during maize development was positively correlated with fumonisin occurrence and levels and negatively with AAL-TA, while moisture events were negatively correlated with fumonisins and positively with AAL-TA. Fumonisin levels were higher in silage harvested at later developmental stages (dough through physiological maturity). Ensiling did not affect toxin concentration nor did agronomic practices (tillage system, inoculant use, or silo type) or silage characteristics (dry matter, pH, or organic acid concentration). This is the first report of AAL-TB in silage and on factors that affect SAM frequency and levels in maize silage.