Brown D.W.,1815 iversity St |
Butchko R.A.E.,1815 iversity St |
Baker S.E.,Pacific Northwest National Laboratory |
Proctor R.H.,1815 iversity St
Fungal Biology | Year: 2012
Fusarium species are ubiquitous in nature, cause a range of plant diseases, and produce a variety of chemicals often referred to as secondary metabolites. Although some fungal secondary metabolites affect plant growth or protect plants from other fungi and bacteria, their presence in grain-based food and feed is more often associated with a variety of diseases in plants and in animals. Many of these structurally diverse metabolites are derived from a family of related enzymes called polyketide synthases (PKSs). A search of genomic sequence of Fusarium verticillioides, Fusarium graminearum, Fusarium oxysporum, and Fusarium solani identified a total of 58 PKS genes. To gain insight into how this gene family evolved and to guide future studies, we conducted phylogenomic and functional domain analyses. The resulting geneaology suggested that Fusarium PKSs represent 34 different groups responsible for synthesis of different core metabolites. The analyses indicate that variation in the Fusarium PKS gene family is due to gene duplication and loss events as well as enzyme gain-of-function due to the acquisition of new domains or of loss-of-function due to nucleotide mutations. Transcriptional analysis indicates that the 16 F. verticillioides PKS genes are expressed under a range of conditions, further evidence that they are functional genes that confer the ability to produce secondary metabolites. © 2011.
Short D.P.G.,Pennsylvania State University |
O'Donnell K.,1815 iversity St |
Zhang N.,Rutgers University |
Juba J.H.,Pennsylvania State University |
Geiser D.M.,Pennsylvania State University
Journal of Clinical Microbiology | Year: 2011
It has been proposed that plumbing systems might serve as a significant environmental reservoir of human-pathogenic isolates of Fusarium. We tested this hypothesis by performing the first extensive multilocus sequence typing (MLST) survey of plumbing drain-associated Fusarium isolates and comparing the diversity observed to the known diversity of clinical Fusarium isolates. We sampled 471 drains, mostly in bathroom sinks, from 131 buildings in the United States using a swabbing method. We found that 66% of sinks and 80% of buildings surveyed yielded at least one Fusarium culture. A total of 297 isolates of Fusarium collected were subjected to MLST to identify the phylogenetic species and sequence types (STs) of these isolates. Our survey revealed that the six most common STs in sinks were identical to the six most frequently associated with human infections. We speculate that the most prevalent STs, by virtue of their ability to form and grow in biofilms, are well adapted to plumbing systems. Six major Fusarium STs were frequently isolated from plumbing drains within a broad geographic area and were identical to STs frequently associated with human infections. Copyright © 2011, American Society for Microbiology. All Rights Reserved.
Dunn R.O.,1815 iversity St
Biofuels | Year: 2015
Biodiesel has several advantages compared to conventional diesel fuel (petrodiesel). Nevertheless, biodiesel has poor cold flow properties that may restrict its use in moderate climates. It is essential that the cold flow properties of biodiesel and its blends with petrodiesel be measured as accurately as possible. This work provides an overview of the important cold flow properties and how they are analyzed. The utility of cloud point (CP), pour point (PP), and cold filter plugging point (CFPP) in evaluating biodiesel at low temperatures is discussed. Advantages and limitations of the experimental methods are evaluated. Finally, the use of sub-ambient differential scanning calorimetry (DSC) in the study of low temperature phase behavior of biodiesel is examined. Copyright © 2015 Taylor & Francis.
Mohamed A.,1815 iversity St |
Xu J.,1815 iversity St |
Singh M.,1815 iversity St
Food Chemistry | Year: 2010
Banana powder (BP) was added to hard-red spring wheat (HRSW) flour intended for yeast-leavened bread formulation. Five different formulations containing 10%, 15%, 20%, 25%, and 30% BP were prepared with varying amounts of base flour, while vital gluten was maintained at 25% in all blends. Based on the added BP amounts only, the prepared bread could deliver 42.87-128.6 mg potassium/30 g of bread (one regular slice) and 0.33-1.00 g of fibre. Although the dough water absorption was increased, due to BP addition, the dough mixing tolerance (MTI) decreased. The bread loaf volume was significantly higher than the control except for the 30% blend, where the loaf volume was similar to the control. Bread staling increased with BP levels due to the high sugar content but, this effect was limited to the first two days of storage. Blends exhibited darker colour due to the high sugar and protein, while the 25% and 30% blends had the lowest percent of freezable water. The amounts of acetic acid extractable proteins from the dry blends and the dough decreased with increase in BP. The linear rheological properties of the control, 10%, and 30% blends exhibited similar viscoelastic solid behaviour, where both G′ and G′′ had plateaus (G′ > G′′) and they were parallel to each other over three decades of the frequency. Blends showed higher moduli values than the control.
Maragos C.M.,1815 iversity St
Journal of AOAC International | Year: 2016
Significant progress has been made in the development of biosensors that can be used to detect low-MW toxins produced by fungi (mycotoxins). The number of formats that have been investigated is impressive and is an indication of the importance attached to finding easy-to-use, accurate, and rapid methods for detecting these toxins in commodities and foods. This review explores the details of multiplexed biosensors based on many formats, including multiplexed immunoassays, suspension arrays, membrane-based devices (flow-through and immunochromatographic), and planar microarrays. Each assay format has its own strengths and areas that need improvement. Certain formats, such as multiplexed immunochromatographic devices, are well developed and relatively easy to use, and in some cases, commercial products are being sold. Others, such as the suspension arrays and microarrays, are laboratory-based assays that, although more complicated, are also more amenable to a larger scale of multiplexing. The diversity of such efforts and the multitude of formats under investigation suggest that multiple solutions will be found to satisfy the need for multiplexed toxin detection.
Brown D.W.,1815 iversity St |
Butchko R.A.E.,1815 iversity St |
Busman M.,1815 iversity St |
Proctor R.H.,1815 iversity St
Fungal Genetics and Biology | Year: 2012
The genus Fusarium is of concern to agricultural production and food/feed safety because of its ability to cause crop disease and to produce mycotoxins. Understanding the genetic basis for production of mycotoxins and other secondary metabolites (SMs) has the potential to limit crop disease and mycotoxin contamination. In fungi, SM biosynthetic genes are typically located adjacent to one another in clusters of co-expressed genes. Such clusters typically include a core gene, responsible for synthesis of an initial chemical, and several genes responsible for chemical modifications, transport, and/or regulation. Fusarium verticillioides is one of the most common pathogens of maize and produces a variety of SMs of concern. Here, we employed whole genome expression analysis and utilized existing knowledge of polyketide synthase (PKS) genes, a common cluster core gene, to identify three novel clusters of co-expressed genes in F. verticillioides. Functional analysis of the PKS genes linked the clusters to production of three known Fusarium SMs, a violet pigment in sexual fruiting bodies (perithecia) and the mycotoxins fusarin C and fusaric acid. The results indicate that microarray analysis of RNA derived from culture conditions that induce differential gene expression can be an effective tool for identifying SM biosynthetic gene clusters. © 2012.
Schnupf U.,1815 iversity St |
Willett J.L.,1815 iversity St |
Momany F.,1815 iversity St
Carbohydrate Research | Year: 2010
Results are presented from density functional molecular dynamics (DFTMD) simulations, based on constant energy dynamics, of glucose and its cyclic form of 6-carbon epimers. Both in vacuo and an implicit solvent method (COSMO) were examined, including simulations of low-energy conformations of each molecule. Analysis of the DFTMD results includes the following: energies averaged over the simulation time, calculated anomeric ratios, hydroxyl and hydroxymethyl rotamer populations, and hydration energies. Hydrogen-bonding networks persistence times were examined, and the effects of solvation on rotamer populations were described. Anomeric ratios calculated from energy optimization of an ensemble of low-energy conformers are compared to those obtained from ensemble averages from molecular dynamics, with dynamics simulations giving populations in best agreement with experimental anomeric ratios. Ensemble results in vacuo were not in agreement with experimental anomeric ratios or hydroxymethyl populations, producing in some cases reversal of the α:β ratios. The difficulty in obtaining correct α:β ratios increases with the number of axial groups; the mono-axial epimers being best represented, epimers with two axial groups being more difficult, and the epimers with three axial hydroxyl groups being most difficult to analyze, the result of a large number of very strong hydrogen-bonding networks that form the ensemble of low-energy conformations in the multi-axial structures.
Johnson E.T.,1815 iversity St
Molecular genetics and genomics : MGG | Year: 2014
The piggyBac transposable element, originally isolated from a virus in an insect cell line, is a valuable molecular tool for transgenesis and mutagenesis of invertebrates. For heterologous transgenesis in a variety of mammals, transfer of the piggyBac transposable element from an ectopic plasmid only requires expression of piggyBac transposase. To determine if piggyBac could function in dicotyledonous plants, a two-element system was developed in tobacco (Nicotiana tabacum) to test for transposable element excision and insertion. The first transgenic line constitutively expressed piggyBac transposase, while the second transgenic line contained at least two non-autonomous piggyBac transposable elements. Progeny from crosses of the two transgenic lines was analyzed for piggyBac excision and transposition. Several progeny displayed excision events, and all the sequenced excision sites exhibited evidence of the precise excision mechanism characteristic of piggyBac transposase. Two unique transposition insertion events were identified that each included diagnostic duplication of the target site. These data indicate that piggyBac transposase is active in a dicotyledonous plant, although at a low frequency.
Brown D.W.,1815 iversity St
Current Genetics | Year: 2011
Killer protein 4 (KP4) is a well studied viral toxin secreted by the maize smut fungus Ustilago maydis that kills sensitive Ustilago strains as well as inhibits Fusarium and plant root growth by inhibiting calcium uptake. Numerous small, cysteine-rich proteins have been shown to play a critical role in fungal-plant-bacterial associations. The discovery of six KP4-like genes in F. verticillioides precipitated efforts to understand their function and evolutionary origin. Analysis of publicly available genomic sequence identified 31 additional KP4-like genes from a range of Ascomycota, a Basidiomycota, and the moss Physcomitrella patens. Sequence comparison and phylogenetic analysis indicate that the viral KP4 and the moss and fungal KP4-like genes evolved from a common ancestor providing evidence for lateral gene transfer between kingdoms. Six genes of the 37 total genes are predicted to encode a protein with two, non-identical KP4-like domains in tandem separated by 29-56 amino acids. The results suggest that two independent events led to the dual-domain KP4 genes present in different lineages of the Ascomycota. Understanding the nature and function of KP4-like proteins in mycotoxin-producing species like Fusarium may help to limit plant diseases and increase food safety and food production. © 2010 Springer-Verlag (outside the USA).
Wiemann P.,University of Munster |
Brown D.W.,1815 iversity St |
Kleigrewe K.,University of Munster |
Bok J.W.,University of Wisconsin - Madison |
And 3 more authors.
Molecular Microbiology | Year: 2010
Besides industrially produced gibberellins (GAs), Fusarium fujikuroi is able to produce additional secondary metabolites such as the pigments bikaverin and neurosporaxanthin and the mycotoxins fumonisins and fusarin C. The global regulation of these biosynthetic pathways is only poorly understood. Recently, the velvet complex containing VeA and several other regulatory proteins was shown to be involved in global regulation of secondary metabolism and differentiation in Aspergillus nidulans. Here, we report on the characterization of two components of the F. fujikuroi velvet-like complex, FfVel1 and FfLae1. The gene encoding this first reported LaeA orthologue outside the class of Eurotiomycetidae is upregulated in δFfvel1 microarray-studies and FfLae1 interacts with FfVel1 in the nucleus. Deletion of Ffvel1 and Fflae1 revealed for the first time that velvet can simultaneously act as positive (GAs, fumonisins and fusarin C) and negative (bikaverin) regulator of secondary metabolism, and that both components affect conidiation and virulence of F. fujikuroi. Furthermore, the velvet-like protein FfVel2 revealed similar functions regarding conidiation, secondary metabolism and virulence as FfVel1. Cross-genus complementation studies of velvet complex component mutants between Fusarium, Aspergillus and Penicillium support an ancient origin for this complex, which has undergone a divergence in specific functions mediating development and secondary metabolism. © 2010 Blackwell Publishing Ltd.