Fargo, ND, United States
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Tsilo T.J.,University of Minnesota | Tsilo T.J.,Small Grain Institute | Simsek S.,North Dakota State University | Ohm J.-B.,Cereal Crops Research Unit | And 3 more authors.
Genome | Year: 2011

Wheat end product quality is determined by a complex group of traits including dough viscoelastic characteristics and bread-making properties. Quantitative trait loci (QTL) mapping and analysis were conducted for endosperm texture, dough-mixing strength, and bread-making properties in a population of 139 (MN99394 × MN98550) recombinant inbred lines that were evaluated at three environments in 2006. Based on the genetic map of 534 loci, six QTL were identified for endosperm texture, with the main QTL on chromosomes 1A (R 2 = 6.6%-17.3%), 5A (R 2 = 6.1%-17.1%), and 5D (R 2 = 15.8%-22%). Thirty-four QTL were identified for eight dough-mixing strength and bread-making properties. Major QTL clusters were associated with the low-molecular weight glutenin gene Glu-A3, the two high-molecular weight glutenin genes Glu-B1 and Glu-D1, and two regions on chromosome 6D. Alleles at these QTL clusters have previously been proven useful for wheat quality, except one of the QTL clusters on chromosome 6D. A QTL cluster on chromosome 6D is one of the novel chromosome regions influencing dough-mixing strength and bread-making properties. The QTL for endosperm texture on chromosomes 1A, 5A, and 5B also influenced flour ash content (12.4%-23.3%), flour protein content (10.5%-12.5%), and flour colour (7.7%-13.5%), respectively. © 2011 Published by NRC Research Press.


McDonald M.C.,ETH Zurich | Oliver R.P.,Curtin University Australia | Friesen T.L.,Cereal Crops Research Unit | Brunner P.C.,ETH Zurich | McDonald B.A.,ETH Zurich
New Phytologist | Year: 2013

Population genetic and phylogenetic studies have shown that Phaeosphaeria nodorum is a member of a species complex that probably shares its center of origin with wheat (Triticum aestivum and Triticum durum). We examined the evolutionary histories of three known necrotrophic effectors (NEs) produced by P. nodorum and compared them with neutral loci. We screened over 1000 individuals for the presence/absence of each effector and assigned each individual to a multi-effector genotype. Diversity at each NE locus was assessed by sequencing c. 200 individuals for each locus. We found significant differences in effector frequency among populations. We propose that these differences reflect the presence/absence of the corresponding susceptibility gene in wheat cultivars. The population harboring the highest sequence diversity was different for each effector locus and never coincided with populations harboring the highest diversity at neutral loci. Coalescent and phylogenetic analyses showed a discontinuous presence of all three NEs among nine closely related Phaeosphaeria species. Only two of the nine species were found to harbor NEs. We present evidence that the three described NEs of P. nodorum were transmitted to its sister species, Phaeosphaeria avenaria tritici 1, via interspecific hybridization. © 2013 The Authors. New Phytologist © 2013 New Phytologist Trust.


Livingston III D.P.,U.S. Department of Agriculture | Livingston III D.P.,North Carolina State University | Henson C.A.,Cereal Crops Research Unit | Henson C.A.,University of Wisconsin - Madison | And 6 more authors.
PLoS ONE | Year: 2013

The crown is the below ground portion of the stem of a grass which contains meristematic cells that give rise to new shoots and roots following winter. To better understand mechanisms of survival from freezing, a histological analysis was performed on rye, wheat, barley and oat plants that had been frozen, thawed and allowed to resume growth under controlled conditions. Extensive tissue disruption and abnormal cell structure was noticed in the center of the crown of all 4 species with relatively normal cells on the outside edge of the crown. A unique visual response was found in oat in the shape of a ring of cells that stained red with Safranin. A tetrazolium analysis indicated that tissues immediately inside this ring were dead and those outside were alive. Fluorescence microscopy revealed that the barrier fluoresced with excitation between 405 and 445 nm. Three dimensional reconstruction of a cross sectional series of images indicated that the red staining cells took on a somewhat spherical shape with regions of no staining where roots entered the crown. Characterizing changes in plants recovering from freezing will help determine the genetic basis for mechanisms involved in this important aspect of winter hardiness.


PubMed | Soybean and Wheat Quality Research Unit and Cereal Crops Research Unit
Type: Journal Article | Journal: Virus genes | Year: 2016

Maize rayado fino virus (MRFV) possesses an open reading frame (ORF43) predicted to encode a 43 kDa protein (p43) that has been postulated to be a viral movement protein. Using a clone of MRFV (pMRFV-US) from which infectious RNA can be produced, point mutations were introduced to either prevent initiation from three potential AUG initiation codons near the 5-end of ORF43 or prematurely terminate translation of ORF43. Inoculation of maize seed via vascular puncture inoculation (VPI) resulted in plants exhibiting symptoms typical of MRFV infection for all mutants tested. Furthermore, corn leafhoppers (Dalbulus maidis) transmitted the virus mutants to healthy plants at a frequency similar to that for wild-type MRFV-US. Viral RNA recovered from plants infected with mutants both prior to and after leafhopper transmission retained mutations blocking ORF43 expression. The results indicate that ORF43 of MRFV is dispensable for both systemic infection of maize and transmission by leafhoppers.


Henson C.A.,Cereal Crops Research Unit | Henson C.A.,University of Wisconsin - Madison | Duke S.H.,University of Wisconsin - Madison | Livingston III D.P.,North Carolina State University
PLoS ONE | Year: 2014

Extensive research has been conducted on cold acclimation and freezing tolerance of fall-sown cereal plants due to their economic importance; however, little has been reported on the biochemical changes occurring over time after the freezing conditions are replaced by conditions favorable for recovery and growth such as would occur during spring. In this study, GC-MS was used to detect metabolic changes in the overwintering crown tissue of oat (Avena sativa L.) during a fourteen day time-course after freezing. Metabolomic analysis revealed increases in most amino acids, particularly proline, 5-oxoproline and arginine, which increased greatly in crowns that were frozen compared to controls and correlated very significantly with days after freezing. In contrast, sugar and sugar related metabolites were little changed by freezing, except sucrose and fructose which decreased dramatically. In frozen tissue all TCA cycle metabolites, especially citrate and malate, decreased in relation to unfrozen tissue. Alterations in some amino acid pools after freezing were similar to those observed in cold acclimation whereas most changes in sugar pools after freezing were not. These similarities and differences suggest that there are common as well as unique genetic mechanisms between these two environmental conditions that are crucial to the winter survival of plants.


Vinje M.A.,University of Wisconsin - Madison | Vinje M.A.,Cereal Crops Research Unit | Willis D.K.,University of Wisconsin - Madison | Willis D.K.,Vegetable Crops Research Unit | And 3 more authors.
Planta | Year: 2011

Two barley (Hordeum vulgare L.) β-amylase genes (Bmy1 and Bmy2) were studied during the late maturation phase of grain development in four genotypes. The Bmy1 and Bmy2 DNA and amino acid sequences are extremely similar. The largest sequence differences are in the introns, seventh exon, and 3′ UTR. Accumulation of Bmy2 mRNA was examined in developing grain at 17, 19, and 21 days after anthesis (DAA). One genotype, PI 296897, had significantly higher Bmy2 RNA transcript accumulation than the other three genotypes at all developmental stages. All four genotypes had Bmy2 mRNA levels decrease from 17 to 19 DAA, and remain the same from 19 to 21 DAA. Levels of Bmy1 mRNA were twenty thousand to over one hundred thousand times more than Bmy2 mRNA levels in genotypes Legacy, Harrington, and Ashqelon at all developmental stages and PI 296897 at 19 and 21 DAA. PI 296897 had five thousand times more Bmy1 mRNA than Bmy2 mRNA at 17 DAA. However, Bmy2 protein was not found at 17 DAA in any genotype. The presence of Bmy2 was immunologically detected at 19 DAA and was present in greater amounts at 21 DAA. Also, Bmy2 protein was found to be stored in mature grain and localized in the soluble fraction. However, Bmy1 protein was far more prevalent than Bmy2 at all developmental stages in all genotypes. Thus, the vast majority of β-amylase activity in developing and mature grain can be attributed to endosperm-specific β-amylase. © 2011 Springer-Verlag (outside the USA).


Duke S.H.,University of Wisconsin - Madison | Henson C.A.,Cereal Crops Research Unit | Henson C.A.,University of Wisconsin - Madison
Journal of the American Society of Brewing Chemists | Year: 2011

This study was conducted to test the following hypotheses: 1) increases in barley wort total sugar concentrations during mashing would be greater than those for malt extract (ME) and more closely reflect increases in wort osmolyte concentrations (OC); and 2) individual wort sugars would correlate better with OC than ME. Malts from 12 barley cultivars were mashed, and wort sugars were quantified at six time points during mashing. Over the initial 55 min of mashing, total sugars for combined cultivars increased 110% compared with 120 and 91% for OC and ME, respectively. At the end of the mashing regime, total sugars for combined cultivars increased to 116% versus 132 and 91-92% for OC and ME, respectively. This indicates starch conversion to sugars was more closely reflected by OC than ME and that after 55 min of mashing sugars and OC continued to increase due to degradation of starch and other malt components to lower molecular weight compounds. In contrast, ME remained relatively constant, supporting the first hypothesis. LSD analysis of data from combined cultivars throughout mashing revealed that both OC and total sugars increased significantly from 55 to 70°C, whereas ME did not, supporting the first hypothesis. Over all time points for combined cultivars, OC correlated better than ME with total sugar concentrations, supporting the second hypothesis. In correlations of individual sugar concentrations with ME and OC for combined and individual cultivars, OC correlated better than ME with all sugars and maltodextrins for combined cultivars and most individual cultivars, supporting the second hypothesis. © 2011 American Society of Brewing Chemists, Inc.


Friesen T.L.,Cereal Crops Research Unit | Faris J.D.,Cereal Crops Research Unit
Canadian Journal of Plant Pathology | Year: 2010

Stagonospora nodorum blotch (SNB) has long been a problem in wheat production areas by affecting both the leaves and glumes of susceptible bread and durum wheat. Resistance to both disease phases has been shown to be complexly inherited and although much effort has gone into the identification and introgression of disease resistance, less than satisfactory progress has been made in producing SNB resistant cultivars. A major pitfall in this process has been the lack of understanding of the underlying mechanism of disease resistance. Recently, we have shown the Stagonospora nodorum-wheat interaction to involve multiple effector proteins also known as host-selective toxins (HSTs) that interact either directly or indirectly with dominant wheat sensitivity/susceptibility gene products to induce disease. Therefore, we have referred to this system as an 'inverse gene-for-gene' interaction (i.e. effector-triggered susceptibility) because the recognition of an effector protein by the host leads to susceptibility rather than resistance as it does in classical gene-for-gene interactions currently referred to as effector-triggered immunity. To date, we have reported five HST-host gene interactions. In each case, toxin sensitivity and susceptibility is controlled by a single dominant gene and in all but one case the interaction is dependent on light. Using quantitative trait loci analysis, the toxin-host gene interactions have been shown to account for 18-95% of the disease variation, highlighting the importance of these interactions. Several unpublished interactions also exist making this a model system for the investigation of the molecular mechanism of necrotrophic disease. © 2010 The Canadian Phytopathological Society.


Duke S.H.,University of Wisconsin - Madison | Vinje M.A.,Cereal Crops Research Unit | Henson C.A.,University of Wisconsin - Madison
Journal of the American Society of Brewing Chemists | Year: 2012

This study was conducted to determine the relationships between patterns of activity of malt amylolytic enzymes (a-amylase, β-amylase, and limit dextrinase) and sugar production in two- and six-row North American cultivars over the course of Congress mashing and to test two hypotheses: 1) that maximal activity of and rates of increase in β-amylase activity during the initial phases of mashing would correlate better than the other amylolytic enzymes with sugar production and 2) that β-amylase intron III allelic variation would have little to no association with sugar production during mashing. Malts of twelve barley cultivars were mashed in a micro-masher and assayed for amylolytic enzyme activities and sugar levels at 6 time points during the 115 min mashing regime. Peak activities of β-amylase were positively and significantly correlated with wort total sugars (r = 0.704, P = 0.011), glucose (r = 0.654, P = 0.021), and maltose (r = 0.780, P = 0.003) and negatively and significantly correlated with maltotetraose (r = -0.830, P = 0.001) and maltopentaose (r = -0.767, P = 0.004). In contrast, with the same comparisons, there were no significant correlations with wort total sugars for α-amylase and limit dextrinase and only a-amylase significantly correlated with some individual sugars (glucose, r = 0.611, P = 0.035; maltotriose, r = 0.594, P = 0.042; maltotetraose, r = -0.772, P = 0.003; maltopentaose, r = -0.728, P = 0.007). Correlations of rates of change in β-amylase activity from five min to the time of maximal activity versus rates of change in total sugars and individual sugars revealed positive and significant correlations with wort total sugars (r = 0.794, P = 0.002), maltose (r = 0.851, P = 0.0004), and maltotriose (r = 0.605, P = 0.038) and significantly negatively correlated with maltotetraose (r = -0.663, P = 0.019) and maltopentaose (r = -0.677, P = 0.016). In contrast, with the same comparisons, there were no significant correlations with wort total sugars or individual sugars versus a-amylase or limit dextrinase. Least significant difference (LSD) analysis revealed that there was no consistent pattern in total wort sugars and the most and least abundant wort sugars and maltodextrins produced during mashing amongst cultivars with Bmy1.a or Bmy1.b intron III alleles. There were no significant differences in cultivars with Bmy1.a or Bmy1.b intron III alleles producing the highest levels of total wort sugars early in mashing, when the bulk of total sugars are produced, (1st 30 min: Bmy1.a [Legacy, Tradition]; Bmy1.b [Harrington]) and at the end of mashing (Bmy1.a [Legacy, Pinnacle, Tradition]; Bmy1.b [Harrington, Merit]). This study supports both of the proposed hypotheses. © 2012 American Society of Brewing Chemists, Inc.


Schmitt M.R.,Cereal Crops Research Unit | Budde A.D.,Cereal Crops Research Unit
Journal of the American Society of Brewing Chemists | Year: 2012

The standard method for determining wort free amino nitrogen content is a long-established but now relatively outdated procedure, calling for the use of test tubes, marbles, several water baths (boiling and 20°C), manual sample transfers, and a conventional spectrophotometer. Currently, many malting quality laboratories avoid use of this cumbersome manual system by adopting automated chemistry analysis systems as their in-house standard method for wort free amino nitrogen analysis. Several recent papers have taken a somewhat different approach to measuring malting quality by adapting the basic analytical method to microtiter plates, microplate readers, and related instrumentation. This streamlines sample handling and increases sample capacity. However, results from the several free amino nitrogen methods published previously have not been directly compared. In this study, we analyzed several malts using the standard ASBC Wort-12 method, our standard Skalar segmented flow analysis chemistry system, and several variations of those two chemistries in microplate formats to compare the results obtained by the different analysis systems. Among these methods, the results from the manual ASBC Wort-12, the automated Skalar system, and a microplate-format assay based on the Skalar reagents agreed well. Surprisingly, however, direct adaptation of the Wort-12 reagents, times, and temperatures to the microplate format yielded inconsistent results. We examined a number of parameters in the microplate-format Wort-12 implementation but were unable to identify a single factor responsible for the divergence of results. From these results, we recommend adapting the chemistry from the Skalar segmented flow analysis system to a microplate format for analysis of wort free amino nitrogen content. © 2012 American Society of Brewing Chemists, Inc.

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