Digital Commons Network^™

Metabolomics-Based Analysis Of Miniature Flask Contents Identifies Tobacco Mixture Use Among The Ancient Maya, Mario Zimmermann, Korey J. Brownstein, Luis Pantoja Díaz, Iliana Ancona Aragón, Scott R. Hutson, Barry Kidder, Shannon Tushingham, David R. Gang

Anthropology Faculty Publications

A particular type of miniature ceramic vessel locally known as “veneneras” is occasionally found during archaeological excavations in the Maya Area. To date, only one study of a collection of such containers successfully identified organic residues through coupled chromatography–mass spectrometry methods. That study identified traces of nicotine likely associated with tobacco. Here we present a more complete picture by analyzing a suite of possible complementary ingredients in tobacco mixtures across a collection of 14 miniature vessels. The collection includes four different vessel forms and allows for the comparison of specimens which had previously formed part of museum exhibitions with recently …

Plant And Microbial Responses To Repeated Cu(Oh)2 Nanopesticide Exposures Under Different Fertilization Levels In An Agro-Ecosystem, Marie Simonin, Benjamin P. Colman, Weiyi Tang, Jonathan D. Judy, Steven M. Anderson, Christina M. Bergemann, Jennifer D. Rocca, Jason M. Unrine, Nicolas Cassar, Emily S. Bernhardt

Plant and Soil Sciences Faculty Publications

The environmental fate and potential impacts of nanopesticides on agroecosystems under realistic agricultural conditions are poorly understood. As a result, the benefits and risks of these novel formulations compared to the conventional products are currently unclear. Here, we examined the effects of repeated realistic exposures of the Cu(OH)₂ nanopesticide, Kocide 3000, on simulated agricultural pastureland in an outdoor mesocosm experiment over 1 year. The Kocide applications were performed alongside three different mineral fertilization levels (Ambient, Low, and High) to assess the environmental impacts of this nanopesticide under low-input or conventional farming scenarios. The effects of Kocide over time were …

Cloning And Characterization Of A Pyrethroid Pesticide Decomposing Esterase Gene, Est3385, From Rhodopseudomonas Palustris Psb-S, Xiangwen Luo, Deyong Zhang, Xuguo Zhou, Jiao Du, Songbai Zhang, Yong Liu

Entomology Faculty Publications

Full length open reading frame of pyrethroid detoxification gene, Est3385, contains 963 nucleotides. This gene was identified and cloned based on the genome sequence of Rhodopseudomonas palustris PSB-S available at the GneBank. The predicted amino acid sequence of Est3385 shared moderate identities (30–46%) with the known homologous esterases. Phylogenetic analysis revealed that Est3385 was a member in the esterase family I. Recombinant Est3385 was heterologous expressed in E. coli, purified and characterized for its substrate specificity, kinetics and stability under various conditions. The optimal temperature and pH for Est3385 were 35 °C and 6.0, respectively. This enzyme could …

Phaeophleospora Vochysiae Savi & Glienke Sp. Nov. Isolated From Vochysia Divergens Found In The Pantanal, Brazil, Produces Bioactive Secondary Metabolites, Daiani C. Savi, Khaled A. Shaaban, Francielly M. W. Gos, Larissa V. Ponomareva, Jon S. Thorson, Chirlei Glienke, Jürgen Rohr

Pharmaceutical Sciences Faculty Publications

Microorganisms associated with plants are highly diverse and can produce a large number of secondary metabolites, with antimicrobial, anti-parasitic and cytotoxic activities. We are particularly interested in exploring endophytes from medicinal plants found in the Pantanal, a unique and widely unexplored wetland in Brazil. In a bio-prospecting study, strains LGMF1213 and LGMF1215 were isolated as endophytes from Vochysia divergens, and by morphological and molecular phylogenetic analyses were characterized as Phaeophleospora vochysiae sp. nov. The chemical assessment of this species reveals three major compounds with high biological activity, cercoscosporin (1), isocercosporin (2) and the new compound 3-(sec-butyl)-6-ethyl-4,5-dihydroxy-2-methoxy-6-methylcyclohex-2-enone (3). …

A New Toolset For Protein Expression And Subcellular Localization Studies In Citrus And Its Application To Citrus Tristeza Virus Proteins, Amit Levy, Choaa El-Mochtar, Chunxia Wang, Michael M. Goodin, Vladimir Orbovic

Plant Pathology Faculty Publications

Background

Transient gene expression is a powerful tool to study gene function in plants. In citrus, Agrobacterium transformation is the method of choice for transient expression studies, but this method does not work efficiently with many gene constructs, and there is a need for a more robust transient expression system in citrus leaves. Biolistic particle delivery is an alternative to Agrobacterium transformation, and in some plants, such as Arabidopsis, gives higher transformation rates in leaf tissues than Agrobacterium.

Results

Here we describe an improved method for gene expression in epidermal cells of citrus leaves, using the Bio-Rad Helios gene-gun. …

Microbial Interactions In The Phyllosphere Increase Plant Performance Under Herbivore Biotic Stress, Muhammad Saleem, Nicole Meckes, Zahida H. Pervaiz, Milton B. Traw

Plant and Soil Sciences Faculty Publications

The phyllosphere supports a tremendous diversity of microbes and other organisms. However, little is known about the colonization and survival of pathogenic and beneficial bacteria alone or together in the phyllosphere across the whole plant life-cycle under herbivory, which hinders our ability to understand the role of phyllosphere bacteria on plant performance. We addressed these questions in experiments using four genetically and biogeographically diverse accessions of Arabidopsis thaliana, three ecologically important bacterial strains (Pseudomonas syringae DC3000, Xanthomonas campestris, both pathogens, and Bacillus cereus, plant beneficial) under common garden conditions that included fungus gnats (Bradysia spp.). …

Regulation Of Pyrroloquinoline Quinone-Dependent Glucose Dehydrogenase Activity In The Model Rhizosphere-Dwelling Bacterium Pseudomonas Putida Kt2440, Ran An, Luke Moe

Plant and Soil Sciences Faculty Publications

Soil-dwelling microbes solubilize mineral phosphates by secreting gluconic acid, which is produced from glucose by a periplasmic glucose dehydrogenase (GDH) that requires pyrroloquinoline quinone (PQQ) as a redox coenzyme. While GDH-dependent phosphate solubilization has been observed in numerous bacteria, little is known concerning the mechanism by which this process is regulated. Here we use the model rhizosphere-dwelling bacterium Pseudomonas putida KT2440 to explore GDH activity and PQQ synthesis, as well as gene expression of the GDH-encoding gene (gcd) and PQQ biosynthesis genes (pqq operon) while under different growth conditions. We also use reverse transcription-PCR to identify transcripts …

Stability Of Antibiotic Resistance Patterns In Agricultural Pastures: Lessons From Kentucky, Usa, Sloane Ritchey, Siva Gandhapudi, Mark Coyne

Plant and Soil Sciences Faculty Publications

No abstract provided.

Novel Model For Multispecies Biofilms That Uses Rigid Gas-Permeable Lenses, Rebecca Peyyala, Sreenatha S. Kirakodu, Jeffrey L. Ebersole, Karen F. Novak

Center for Oral Health Research Faculty Publications

Oral biofilms comprise complex multispecies consortia aided by specific inter- and intraspecies interactions occurring among commensals and pathogenic bacterial species. Oral biofilms are primary initiating factors of periodontal disease, although complex multifactorial biological influences, including host cell responses, contribute to the individual outcome of the disease. To provide a system to study initial stages of interaction between oral biofilms and the host cells that contribute to the disease process, we developed a novel in vitro model system to grow biofilms on rigid gas-permeable contact lenses (RGPLs), which enable oxygen to permeate through the lens material. Bacterial species belonging to early- …

Soil Organic Matter Fractions And Aggregate Distribution In Response To Tall Fescue Stands, I. P. Handayani, Mark S. Coyne, R. S. Tokosh

Plant and Soil Sciences Faculty Publications

The study was conducted to evaluate the influences of tall fescue management on soil organic matter fractions and macro- and microaggregate distribution. Soil samples were collected from four paired adjacent fields consisting of five years of tall fescue mono and poly stands in Western Kentucky. Soil samples from 0 to 15 cm and 15 to 30 cm soil depths were analyzed for soil organic C and N, particulate organic matter C (POM-C) and N (POM-N), macro- and micro aggregate distribution and C-associated with macro- and micro- aggregates. Significant effects were observed between stands for all the properties, except total C, …

Multimeric Hemicellulases Facilitate Biomass Conversion, Zhanmin Fan, Kurt Wagschal, Wei Chen, Michael D. Montross, Charles C. Lee, Ling Yuan

Plant and Soil Sciences Faculty Publications

Two highly active trifunctional hemicellulases were constructed by linking the catalytic portion of a xylanase with an arabinofuranosidase and a xylosidase, using either flexible peptide linkers or linkers containing a cellulose-binding domain. The multifunctional enzymes retain the parental enzyme properties and exhibit synergistic effects in hydrolysis of natural xylans and corn stover.

Soil Microbial Community Response To Hexavalent Chromium In Planted And Unplanted Soil, Ioannis Ipsilantis, Mark S. Coyne

Plant and Soil Sciences Faculty Publications

Theories suggest that rapid microbial growth rates lead to quicker development of metal resistance. We tested these theories by adding hexavalent chromium [Cr(VI)] to soil, sowing Indian mustard (Brassica juncea), and comparing rhizosphere and bulk soil microbial community responses. Four weeks after the initial Cr(VI) application we measured Cr concentration, microbial biomass by fumigation extraction and soil extract ATP, tolerance to Cr and growth rates with tritiated thymidine incorporation, and performed community substrate use analysis with BIOLOG GN plates. Exchangeable Cr(VI) levels were very low, and therefore we assumed the Cr(VI) impact was transient. Microbial biomass was reduced …

Effects Of Commercial Diazinon And Imidacloprid On Microbial Urease Activity In Soil And Sod, C. W. Ingram, Mark S. Coyne, David W. Williams

Plant and Soil Sciences Faculty Publications

Diazinon [O,O-diethyl O-2-isopropyl-6-methyl(pyrimidine-4-yl) phosphorothioate] and imidacloprid [1-(1-[6-chloro-3-pyridinyl]methyl)-N-nitro-2-imidazolidinimine] are applied to lawns for insect control simultaneously with nitrogenous fertilizers such as urea, but their potential effect on urease activity and nitrogen availability in turfgrass management has not been evaluated. Urease activity in enzyme assays, washed cell assays, and soil slurries was examined as a function of insecticide concentration. Intact cores from field sites were used to assess the effect of insecticide application on urease activity in creeping bentgrass (Agrostis palustris Huds.) and bluegrass (Poa pratensis L.) sod. Bacterial urease fromBacillus pasteurii and plant …

Fertilizer, Tillage, And Dairy Manure Contributions To Nitrate And Herbicide Leaching, C. S. Stoddard, John H. Grove, Mark S. Coyne, William O. Thom

Plant and Soil Sciences Faculty Publications

Few studies have examined the water quality impact of manure use in no-tillage systems. A lysimeter study in continuous corn (Zea mays L.) was performed on Maury silt loam (fine, mixed, semiactive, mesic Typic Paleudalf) to evaluate the effect(s) of tillage (no-till [NT] and chisel-disk [CD]), nitrogen fertilizer rate (0 and 168 kg N ha⁻¹), and dairy manure application timing (none, spring, fall, or fall plus spring) on NO₃–N, atrazine (2-chloro-4-ethylamino-6-isopropylamino-s-triazine), and alachlor [2-chloro-2′-6′-diethyl-N-(methoxymethyl)acetanilide] concentrations in leachate collected at a 90-cm depth. Herbicides were highest immediately after application, declining to less …

Mortality Of Escherichia Coli O157:H7 In Two Soils With Different Physical And Chemical Properties, D. N. Mubiru, Mark S. Coyne, John H. Grove

Plant and Soil Sciences Faculty Publications

Wild and domesticated animals can harbor a pathogenic Escherichia coli strain designated as O157:H7. Potential health problems could occur if strain O157:H7 is a more robust survivor in defecated waste than commonly used indicator bacteria. A laboratory study was conducted to assess E. coli O157:H7 survival relative to a nonpathogenie E. coli strain in two soils with different physical and chemical characteristics. Bacteria in the inoculated soils were enumerated on a weekly basis for 8 wk using a most probable number (MPN) technique. First-order decay models were used to describe bacteria mortality in the soils. Decay series were described slightly …

Fecal Bacteria Survival And Infiltration Through A Shallow Agricultural Soil: Timing And Tillage Effects, C. S. Stoddard, Mark S. Coyne, John H. Grove

Plant and Soil Sciences Faculty Publications

Human and livestock exposure to fecal pathogens via contaminated surface or groundwater is an important water quality concern for soils receiving animal wastes. The effects of manure application timing (spring or fall application) and soil management (no-tillage or conservation tillage) on fecal bacteria infiltration through shallow karst soils in central Kentucky (the Bluegrass region) have not been evaluated. We performed a field experiment to measure fecal coliforms and fecal streptococci in leachate from dairy manure-amended no-tillage and conservation tillage soils. Manure significantly increased fecal bacteria in leachate compared with unmanured treatments. After manure application, the leachate that collected in zero-tension …

Fecal Coliform Transport Through Intact Soil Blocks Amended With Poultry Manure, S. W. Mcmurry, Mark S. Coyne, E. Perfect

Plant and Soil Sciences Faculty Publications

Poultry production in Kentucky increased almost 200% between 1991 and 1995. Their waste is typically land applied, and fecal pathogen runoff and infiltration may cause nonpoint source groundwater pollution. We looked at the preferential flow of fecal coliforms through undisturbed soil blocks since fecal bacteria typically infiltrate the soil profile to contaminate groundwater. Poultry manure was uniformly distributed on top of sod-covered or tilled (upper 12.5 cm) soil blocks and the blocks were irrigated. Drainage was collected in 100 uniformly spaced cells beneath each block and analyzed for fecal coliform content and drainage volume. The spatial distribution of drainage and …

Effect Of Sediment Particle Size And Temperature On Fecal Bacteria Mortality Rates And The Fecal Coliform/Fecal Streptococci Ratio, J. M. Howell, Mark S. Coyne, P. L. Cornelius

Plant and Soil Sciences Faculty Publications

Extended survival of fecal bacteria in sediment can obscure the source and extent of fecal contamination in agricultural settings. The variability in fecal coliform/fecal streptococci (FC/FS) ratios with time and discrepancies between observable fecal sources and measured FC/FS ratios in shallow surface water from agricultural watersheds may be explained by examining FC and FS mortality rates in response to ambient temperature and sediment particle size. We measured FC and FS mortality rates at three different temperatures and in three feces-amended sediments with different particle size in a laboratory study. In controlled conditions, using physiological saline to reduce cell death by …

Nitrous Oxide Loss From Poultry Manure-Amended Soil After Rain, Mark S. Coyne, A. Villalba, Robert L. Blevins

Plant and Soil Sciences Faculty Publications

Land application of poultry wastes in Kentucky will increase as the broiler industry grows. If poultry manure stimulates N₂O loss from soil it will reduce the fertilizer N value of this waste. In contrast, stimulated N₂O loss in grass filter strips receiving the runoff from manured fields could help reduce contamination of surface water by NO⁻₃. Our objectives were to determine (i) if poultry manure stimulated N₂O loss in soil after rainfall and (ii) if there was an edge-of-field effect on N₂O loss in grass filters intercepting runoff from …

Fecal Bacteria In Agricultural Waters Of The Bluegrass Region Of Kentucky, J. M. Howell, Mark S. Coyne, P. L. Cornelius

Plant and Soil Sciences Faculty Publications

Agricultural runoff influenced by nonpoint pollution frequently exceeds the USEPA standards for bacterial contamination of primary contact water (200 fecal coliforms/100 mL). Few studies have evaluated the effect of cattle (Bos taurus) grazing on fecal contamination of ground water in the karst topography of central Kentucky. Our objectives were to: (i) observe the extent and pattern of fecal bacteria in agricultural waters from two central Kentucky watersheds; (ii) determine if monthly sampling accurately assessed the extent and variability of fecal contamination; and (iii) assess the fecal coliform/fecal streptococci ratio (FC/FS) as an indicator of fecal bacteria source. Springs, …

Nitrous Oxide Flux From Poultry-Manured Erosion Plots And Grass Filters After Simulated Rain, Mark S. Coyne, R. A. Gilfillen, Robert L. Blevins

Plant and Soil Sciences Faculty Publications

Adding carbon-rich materials to fields, like manure, may enhance denitrification. Grass filters, which are used to trap surface runoff from these fields, may also provide a carbon-rich environment that favors water infiltration and denitrification. Nitrous oxide (N₂O) may be evolved these settings. It is a radiatively important trace gas and intermediate in the denitrification pathway and several other microbial processes. We measured N₂O flux, after simulated rain, using a soil cover technique in poultry-manured plots and grass filters receiving their runoff. Intact soil cores were used to relate the N₂O flux to the denitrification …

Frequency Of Mug Negative Escherichia Coli In Kentucky Groundwater Samples, Mark S. Coyne, J. C. Shuler

Plant and Soil Sciences Faculty Publications

MUG negative Escherichia coli are a small fraction (2.5%) of the total E. coli in Kentucky groundwater samples. It is unlikely that they alone will cause a significant potential to underestimate fecal contamination using MUG as the primary criterion for that assessment. An unresolved question is how effectively MUG-based, defined-substrate tests address false negative water samples containing MUG positive E. coli.

Sunlight Induced Movement Of Planktonic Organisms And Their Relationships To Water Movements, John Calkins, Ed Colley, Jay Wallingford, Carol Hulsey, Kathy Lohr, Marc Bolling

KWRRI Research Reports

Previous studies have shown the planktonic organisms of various natural water bodies to be quite sensitive to the levels of solar ultraviolet radiation incident at the water surface in summer. It has been assumed that the majority of the phytoplankton are simply carried about in the mass of the water which they inhabit with little control of their position or sunlight exposure. Photosynthetic microorganisms must strike a delicate balance with regard to solar radiation; they must obtain enough visible light for adequate photosynthesis while avoiding an excessive exposure to the injurious solar UV-B radiation.

It is proposed that the circulation …