Fargo, ND, United States
Fargo, ND, United States

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Anderson J.V.,Biosciences Research Laboratory | Tedrow T.,Biosciences Research Laboratory | Tedrow T.,Kansas State University | Hulke B.,U.S. Department of Agriculture | Kennedy A.C.,Washington State University
Journal of Agricultural and Food Chemistry | Year: 2010

Incubation of dormant wild oat (Avena fatua L., isoline M73) caryopses for 1-3 days with Fusarium avenaceum seed-decay isolate F.a.1 induced activity of the plant defense enzyme polyphenol oxidase (PPO). Both extracts and leachates obtained from F.a.1-treated caryopses had decreased abundance of an ̃57 kDa antigenic PPO and increased abundance of antigenic PPOs ranging from ̃52 to 14 kDa, as compared to extracts and leachates from untreated caryopsis. Leachates from caryopsis incubated for 2 days with F.a.1 also had 5.1- and 7.5-fold more total PPO activity/g fwt and specific activity, respectively. Fractionation of leachate proteins by ion-exchange chromatography associated the majority of PPO activity with an ̃36 kDa protein from untreated caryopses and ̃36, 25, and 24 kDa proteins from F.a.1-treated caryopses. Predicted peptide sequences obtained from high-performance liquid chromatography-tandem mass spectrometry analyses indicated that the ̃57 and 36 kDa wild oat proteins had a strong similarity to wheat PPO. However, the 25 and 24 kDa proteins were most similar to a Chitinase and oxalate oxidase, respectively. Our results indicate that F.a.1-induced activation of latent PPO in wild oat caryopsis likely involves a cleavage mechanism allowing activated PPOs to be readily mobilized into their surrounding environment. © 2010 American Chemical Society.


Gallagher R.S.,Pennsylvania State University | Ananth R.,Pennsylvania State University | Granger K.,Pennsylvania State University | Bradley B.,Pennsylvania State University | And 2 more authors.
Journal of Agricultural and Food Chemistry | Year: 2010

The objective of this research was to identify and quantify the phenolic and short-chained aliphatic organic acids present in the seeds of three wild-type populations of wild oat and compare these results to the chemical composition of seeds from two commonly utilized wild oat isolines (M73 and SH430). Phenolic acids have been shown to serve as germination inhibitors, as well as protection for seeds from biotic and abiotic stress factors in other species, whereas aliphatic organic acids have been linked to germination traits and protection against pathogens. Wild oat populations were grown under a "common garden" environment to remove maternal variation, and the resulting seeds were extracted to remove the readily soluble and chemically bound phenolic and aliphatic organic acid components. Compounds were identified and quantified using gas chromatography-mass spectrometry. Ferulic and p-coumaric acid comprised 99% of the total phenolic acids present in the seeds, of which 91% were contained in the hulls and 98% were in the chemically bound forms. Smaller quantities of OH benzoic and vanillic acid were also detected. Soluble organic acids concentrations were higher in the M73 isoline compared to SH430, suggesting that these chemical constituents could be related to seed dormancy. Malic, succinic, fumaric and azelaic acid were the dominant aliphatic organic acids detected in all seed and chemical fractions. © 2009 American Chemical Society.


PubMed | Biosciences Research Laboratory, Chungbuk National University, Southern Plains Agricultural Research Center and Texas A&M University
Type: | Journal: Food chemistry | Year: 2016

Lipolysis and biohydrogenation in ruminal animals promote the accumulation of saturated fatty acids in their meat and milk. Antibodies were generated against key ruminal lipase contributors Anaerovibrio lipolyticus, Butyrivibrio fibrisolvens, Propionibacterium avidum and acnes. An anti-Pseudomonas lipase antibody was generated to determine if an antibody against a purified protein would be more effective. Each bacterium was cultured and assayed without or with increasing levels of each antibody. Butyrivibrio fibrisolvens H17C also participates in biohydrogenation and therefore the antibody was tested to determine if it could effectively reduce biohydrogenation. Butyrivibrio fibrisolvens was assayed without and with the anti-B. fibrisolvens antibody and linoleic or -linolenic acid. All antibodies were effective at reducing lipolysis with the anti-Pseudomonas lipase averaging a 78% reduction. The anti-B. fibrisolvens showed a tendency for a reduction (P=0.0713) in biohydrogenation products of -linolenic acid. Results demonstrate that lipolysis and biohydrogenation can be immunologically inhibited in vitro.


Liu Z.,North Dakota State University | El-Basyoni I.,University of Nebraska - Lincoln | Kariyawasam G.,North Dakota State University | Zhang G.,Kansas State University | And 7 more authors.
Plant Disease | Year: 2015

Tan spot and Stagonospora nodorum blotch (SNB), often occurring together, are two economically significant diseases of wheat in the Northern Great Plains of the United States. They are caused by the fungi Pyrenophora tritici-repentis and Parastagonospora nodorum, respectively, both of which produce multiple necrotrophic effectors (NE) to cause disease. In this work, 120 hard red winter wheat (HRWW) cultivars or elite lines, mostly from the United States, were evaluated in the greenhouse for their reactions to the two diseases as well as NE produced by the two pathogens. One P. nodorum isolate (Sn4) and four Pyrenophora tritici-repentis isolates (Pti2, 331-9, DW5, and AR CrossB10) were used separately in the disease evaluations. NE sensitivity evaluation included ToxA, Ptr ToxB, SnTox1, and SnTox3. The numbers of lines that wer rated highly resistant to individual isolates ranged from 11 (9%) to 30 (25%) but only six lines (5%) were highly resistant to all isolates, indicating limited sources of resistance to both diseases in the U.S. adapted HRWW germplasm. Sensitivity to ToxA was identified in 83 (69%) of the lines and significantly correlated with disease caused by Sn4 and Pti2, whereas sensitivity to other NE was present at much lower frequency and had no significant association with disease. As expected, association mapping located ToxA and SnTox3 sensitivity to chromosome arm 5BL and 5BS, respectively. A total of 24 potential quantitative trait loci was identified with −log (P value) > 3.0 on 12 chromosomes, some of which are novel. This work provides valuable information and tools for HRWW production and breeding in the Northern Great Plains. © 2015 The American Phytopathological Society.


Chao W.S.,Biosciences Research Laboratory | Dogramaci M.,Biosciences Research Laboratory | Foley M.E.,Biosciences Research Laboratory | Horvath D.P.,Biosciences Research Laboratory | Anderson J.V.,Biosciences Research Laboratory
PLoS ONE | Year: 2012

Quantitative real-time polymerase chain reaction (qRT-PCR) is the most important tool in measuring levels of gene expression due to its accuracy, specificity, and sensitivity. However, the accuracy of qRT-PCR analysis strongly depends on transcript normalization using stably expressed reference genes. The aim of this study was to find internal reference genes for qRT-PCR analysis in various experimental conditions for seed, adventitious underground bud, and other organs of leafy spurge. Eleven candidate reference genes (BAM4, PU1, TRP-like, FRO1, ORE9, BAM1, SEU, ARF2, KAPP, ZTL, and MPK4) were selected from among 171 genes based on expression stabilities during seed germination and bud growth. The other ten candidate reference genes were selected from three different sources: (1) 3 stably expressed leafy spurge genes (60S, bZIP21, and MD-100) identified from the analyses of leafy spurge microarray data; (2) 3 orthologs of Arabidopsis "general purpose" traditional reference genes (GAPDH_1, GAPDH_2, and UBC); and (3) 4 orthologs of Arabidopsis stably expressed genes (UBC9, SAND, PTB, and F-box) identified from Affymetrix ATH1 whole-genome GeneChip studies. The expression stabilities of these 21 genes were ranked based on the CT values of 72 samples using four different computation programs including geNorm, Normfinder, BestKeeper, and the comparative ΔCT method. Our analyses revealed SAND, PTB, ORE9, and ARF2 to be the most appropriate reference genes for accurate normalization of gene expression data. Since SAND and PTB were obtained from 4 orthologs of Arabidopsis, while ORE9 and ARF2 were selected from 171 leafy spurge genes, it was more efficient to identify good reference genes from the orthologs of other plant species that were known to be stably expressed than that of randomly testing endogenous genes. Nevertheless, the two newly identified leafy spurge genes, ORE9 and ARF2, can serve as orthologous candidates in the search for reference genes from other plant species.


PubMed | Biosciences Research Laboratory
Type: Journal Article | Journal: Journal of environmental quality | Year: 2011

The excreted estrogen rate of all livestock in the United States is estimated at 134 kg d. The influence of manure treatment on the fate of estrogens is critical in deciding the recycling of over 300 million dry tons of livestock produced annually. The effects of two common manure management practices, heated composting and ambient temperature decomposition, on the fate of 17-estradiol in poultry litter were determined. A mixture of poultry litter, wood chips, and straw was amended with [C]17-estradiol and allowed to undergo decomposition with a laboratory-scale heated composter (HC) or room temperature incubation (RTI) for 24 d. Radiolabel in the finished products was fractionated into water-extractable, acetone-extractable, nonextractable, and mineralized fractions. Total 17-estradiol radioactive residues in the HC and RTI ( = 2) treatments were not different ( > 0.05), except that statistically less 17-estradiol was mineralized to CO during HC than RTI (1.1 vs. 10.0% for HC and RTI, respectively). Estrone was the major degradation product in extracts of HC and RTI treatments as determined by liquid chromatography/mass spectrometry analyses. The nonextractable residues indicated no quantitative differences among the humins between the treatments. An estimated 3% of the fortified estrogenicity remained after HC treatment, and 15% of the fortified estrogenicity remained after RTI treatment. If reduction of water-removable, biologically active 17-estradiol is the treatment goal, then HC treatment would be slightly preferred over ambient temperature degradation. However, unmanaged, ambient temperature litter piles are less costly and time consuming for food animal producers and result in greater mineralization and similar immobilization of estradiol.


PubMed | Biosciences Research Laboratory
Type: Journal Article | Journal: PloS one | Year: 2012

Quantitative real-time polymerase chain reaction (qRT-PCR) is the most important tool in measuring levels of gene expression due to its accuracy, specificity, and sensitivity. However, the accuracy of qRT-PCR analysis strongly depends on transcript normalization using stably expressed reference genes. The aim of this study was to find internal reference genes for qRT-PCR analysis in various experimental conditions for seed, adventitious underground bud, and other organs of leafy spurge. Eleven candidate reference genes (BAM4, PU1, TRP-like, FRO1, ORE9, BAM1, SEU, ARF2, KAPP, ZTL, and MPK4) were selected from among 171 genes based on expression stabilities during seed germination and bud growth. The other ten candidate reference genes were selected from three different sources: (1) 3 stably expressed leafy spurge genes (60S, bZIP21, and MD-100) identified from the analyses of leafy spurge microarray data; (2) 3 orthologs of Arabidopsis general purpose traditional reference genes (GAPDH_1, GAPDH_2, and UBC); and (3) 4 orthologs of Arabidopsis stably expressed genes (UBC9, SAND, PTB, and F-box) identified from Affymetrix ATH1 whole-genome GeneChip studies. The expression stabilities of these 21 genes were ranked based on the C(T) values of 72 samples using four different computation programs including geNorm, Normfinder, BestKeeper, and the comparative C(T) method. Our analyses revealed SAND, PTB, ORE9, and ARF2 to be the most appropriate reference genes for accurate normalization of gene expression data. Since SAND and PTB were obtained from 4 orthologs of Arabidopsis, while ORE9 and ARF2 were selected from 171 leafy spurge genes, it was more efficient to identify good reference genes from the orthologs of other plant species that were known to be stably expressed than that of randomly testing endogenous genes. Nevertheless, the two newly identified leafy spurge genes, ORE9 and ARF2, can serve as orthologous candidates in the search for reference genes from other plant species.

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