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Efficient And Stable Expression Of Gfp Through Wheat Streak Mosaic Virus-Based Vectors In Cereal Hosts Using A Range Of Cleavage Sites: Formation Of Dense Fluorescent Aggregates For Sensitive Virus Tracking, Satyanarayana Tatineni, Anthony J. Mcmechan, Gary L. Hein, Roy French

Department of Plant Pathology: Faculty Publications

A series of Wheat streak mosaic virus (WSMV)-based expression vectors were developed by engineering a cycle 3 GFP (GFP) cistron between P1 and HC-Pro cistrons with several catalytic/cleavage peptides at the Cterminus of GFP. WSMV-GFP vectors with the Foot-and-mouth disease virus 1D/2A or 2A catalytic peptides cleaved GFP from HC-Pro but expressed GFP inefficiently. WSMV-GFP vectors with homologous NIa-Pro heptapeptide cleavage sites did not release GFP from HC-Pro, but efficiently expressed GFP as dense fluorescent aggregates. However, insertion of one or two spacer amino acids on either side of NIb/CP heptapeptide cleavage site or deletion in HC-Pro cistron improved processing …

Another Really, Really Big Virus, James L. Van Etten

Department of Plant Pathology: Faculty Publications

Viruses with genomes larger than 300 kb and up to 1.2 Mb, which encode hundreds of proteins, are being discovered and characterized with increasing frequency. Most, but not all, of these large viruses (often referred to as giruses) infect protists that live in aqueous environments. Bioinformatic analyses of metagenomes of aqueous samples indicate that large DNA viruses are quite common in nature and await discovery. One issue that is perhaps not appreciated by the virology community is that large viruses, even those classified in the same family, can differ significantly in morphology, lifestyle, and gene complement. This brief commentary, which …

Motus, Morphology, And Biodiversity Estimation: A Case Study Using Nematodes Of The Suborder Criconematina And A Conserved 18s Dna Barcode, Thomas O. Powers, Timothy Harris, Rebecca Higgins, Peter Mullin, Lisa Sutton, Kirsten Powers

Department of Plant Pathology: Faculty Publications

DNA barcodes are increasingly used to provide an estimate of biodiversity for small, cryptic organisms like nematodes. Nucleotide sequences generated by the barcoding process are often grouped, based on similarity, into molecular operational taxonomic units (MOTUs). In order to get a better understanding of the taxonomic resolution of a 3’ 592-bp 18S rDNA barcode, we have analyzed 100 MOTUs generated from 214 specimens in the nematode suborder Criconematina. Previous research has demonstrated that the primer set for this barcode reliably amplifies all nematodes in the Phylum Nematoda. Included among the Criconematina specimens were 25 morphologically described species representing 12 genera. …

Presence Of Fusarium Spp. In Air And Soil Associated With Sorghum Fields, Deanna L. Funnell-Harris, Jeffrey F. Pedersen

Department of Plant Pathology: Faculty Publications

Sorghum grain, valuable for feed, food, and bioenergy, can be colonized by several Fusarium spp.; therefore, it was of interest to identify possible sources of conidia. Analysis of air and soil samples provided evidence for the presence of propagules from Fusarium genotypes that may cause grain infections. Soil population estimates of members of the Gibberella fujikuroi species complex, that includes sorghum pathogens and other Fusarium spp., suggested that adequate inoculum for systemic infections was present. Conidia in air samples within two sorghum fields were collected by passive trapping for 2 years. Subsampled Fusarium isolates indicated that numbers of G. fujikuroi …

First Report Of Goss's Bacterial Wilt And Leaf Blight (Clavibacter Michiganensis Subsp. Nebraskensis) Of Corn In Texas, K. A. Korus, A. D. Timmerman, R. D. French-Monar, T. A. Jackson

Department of Plant Pathology: Faculty Publications

In September 2009, the University of Nebraska–Lincoln Plant and Pest Diagnostic Clinic received leaf samples of hybrid corn (Zea mays L.) displaying long, necrotic lesions with wavy margins. The lesions had discontinuous water-soaked spots that are indicative of Goss's bacterial wilt and leaf blight. The symptomatic leaves were submitted from Dallam County, located in the Texas Panhandle (northwest Texas). According to the USDA Farm Service Agency and the National Agricultural Statistics Service, in 2009 Dallam County had 54,025 ha planted to corn. This is approximately 19% of the total corn planted in the 26 counties in the Texas Panhandle …

Effects Of Integrating Cultivar Resistance And Fungicide Application On Fusarium Head Blight And Deoxynivalenol In Winter Wheat, Stephen N. Wegulo, William W. Bockus, John Hernandez Nopsa, Erick D. De Wolf, Kent M. Eskridge, Kamaranga H. S. Peiris, Floyd E. Dowell

Department of Plant Pathology: Faculty Publications

Fusarium head blight (FHB) or scab, incited by Fusarium graminearum, can cause significant economic losses in small grain production. Five field experiments were conducted from 2007 to 2009 to determine the effects on FHB and the associated mycotoxin deoxynivalenol (DON) of integrating winter wheat cultivar resistance and fungicide application. Other variables measured were yield and the percentage of Fusarium-damaged kernels (FDK). The fungicides prothioconazole + tebuconazole (formulated as Prosaro 421 SC) were applied at the rate of 0.475 liters/ha, or not applied, to three cultivars (experiments 1 to 3) or six cultivars (experiments 4 and 5) differing in their …

Characterization Of Three-Dimensional Spatial Aggregation And Association Patterns Of Brown Rot Symptoms Within Intensively Mapped Sour Cherry Trees, Sydney E. Everhart, Ashley Askew, Lynne Seymour, Imre J. Holb, Harald Scherm

Department of Plant Pathology: Faculty Publications

Characterization of spatial patterns of plant disease can provide insights into important epidemiological processes such as sources of inoculum, mechanisms of dissemination, and reproductive strategies of the pathogen population. While two-dimensional patterns of disease (among plants within fields) have been studied extensively, there is limited information on three-dimensional patterns within individual plant canopies. Reported here are the detailed mapping of different symptom types of brown rot (caused by Monilinia laxa) in individual sour cherry tree (Prunus cerasus) canopies, and the application of spatial statistics to the resulting data points to de-termine patterns of symptom aggregation and association. …

Floristic And Environmental Characteristics Of Sub-Boreal Peatlands Of Minnesota And Western Wisconsin, Kevin Douglas Clement

All Graduate Theses, Dissertations, and Other Capstone Projects

Peatland communities in the sub-boreal region of Minnesota and western Wisconsin were characterized by floristic composition, structure and environmental characteristics to determine the natural variation among peatland communities in the region and their shared associations with the boreal and temperate peatlands of North America. Floristic classification revealed five, distinct peatland community types, identified as Sphagnum Bogs, Herbaceous Fens, Forested Fens, Rich/Calcareous Fens and Phalaris anrundinacea dominated peatlands. Differences among sub-boreal peatlands largely reflected changes in the significant indicator and dominant plant species identified, which were found to exhibit high fidelities to a particular peatland community type. Floristic variations among sub-boreal …

Distributions And Abundances Of Microstegium Vimineum Along Forest Roadsides At The Grassy Hill Natural Area, Franklin County, Virginia, Gregory D. Turner

Virginia Journal of Science

In summer 2005, plots were surveyed along roads passing through forest habitats at the Grassy Hill Natural Area Preserve in Franklin County, Virginia to assess the distributions and abundances of Microstegium vimineum in transects located at increasing distances away from roadsides into forest interiors. Across plots, Microstegium was encountered almost exclusively in roadside transects, where abundances were relatively high. While forest composition and topographic features were similar across plots, percent canopy cover and leaf litter depth were greater in interior compared to roadside transects due to undisturbed tree canopies and ground cover located in interior plot areas. Results imply that …

Role Of The Sexual Cycle In Development Of Genotypic And Phenotypic Diversity In Gibberella Zeae, Sladana Bec

Theses and Dissertations--Plant Pathology

Gibberella zeae (anamorph Fusarium graminearum) is a homothallic ascomycete pathogen that is responsible for causing Fusarium head blight (FHB) of wheat and small grains. In addition to causing a reduction in yield, harvested grain is frequently contaminated with trichothecene mycotoxins that are harmful for human and animal health. Use of wheat varieties with resistance to FHB is an important strategy to lower its impact. In order to produce varieties with durable resistance, we must understand the origin and degree of genetic diversity present in the pathogen population. In my research, I focused my efforts on an investigation of the …

Saplings 2011, Sean M. Goins, Joseph Braner, Amy L. Myers, Charlie Jackson, Julia I. Chapman, Ryan W. Mcewan

Data Files: Drew Woods Permanent Plots

Sapling data (woody species >50 centimeters in height and <2.5 cm diameter at breast height) collected in 2011 at Drew Woods State Nature Preserve in Darke County, OH. Data key is provided in a supplemental file.

Overstory 2011, Sean M. Goins, Joseph Braner, Amy L. Myers, Charlie Jackson, Albert J. Burky, Julia I. Chapman, Ryan W. Mcewan

Data Files: Drew Woods Permanent Plots

Overstory data (woody species ≥2.5 centimeters in diameter at breast height) collected in 2011 at Drew Woods State Nature Preserve in Darke County, OH. Data key is provided in a supplemental file.

Role Of Lipids In Tombusvirus Replication, Monika Sharma

University of Kentucky Doctoral Dissertations

Positive-strand RNA virus group are the most abundant among viruses affecting plants and animals. To successfully achieve replication, these viruses usurp or co-opt host proteins. To facilitate the discovery of host factors involved in Tomato bushy stunt virus (TBSV), yeast has been developed as a surrogate model host. Genome-wide approaches covering 95% of yeast genes, has revealed approximately hundred factors that could affect virus replication. Among the identified host factors, there are fourteen yeast genes, which affect/regulate lipid metabolism of the host.

One of the identified host gene is ERG25, which is an important factor for sterol biosynthesis pathway, affecting …

Characterization Of G10h Promoter And Isolation Of Wrky Transcription Factors Involved In Catharanthus Terpenoid Indole Alkaloid Biosynthesis Pathway, Nitima Suttpanta

University of Kentucky Doctoral Dissertations

Catharanthus roseus produces a large array of terpenoid indole alkaloids (TIAs) that are an important source of natural or semi-synthetic anticancer drugs. Biosynthesis of TIAs is tissue-specific and induced by certain phytohormones and fungal elicitors, indicating the involvement of a complex transcriptional control network. However, the transcriptional regulation of the TIA pathway is poorly understood. This study reports the isolation and characterization of the G10H promoter and two WRKY transcription factors regulating TIA biosynthesis.

Geraniol 10-hydroxylase (G10H) controls the first committed step in the biosynthesis of terpenoid indole alkaloids (TIA). The C. roseus G10H promoter sequence was isolated by a …

Estimation Of Surface Soil Organic Matter Using A Ground-Based Active Sensor And Aerial Imagery, D. F. Roberts, V. I. Adamchuk, J. F. Shanahan, R. B. Ferguson, J. S. Schepers

Department of Agronomy and Horticulture: Faculty Publications

Active canopy sensors are currently being studied as a tool to assess crop N status and direct in-season N applications. The objective of this study was to use a variety of strategies to evaluate the capability of an active sensor and a wide-band aerial image to estimate surface soil organic matter (OM). Grid soil samples, active sensor reflectance and bare soil aerial images were obtained from six fields in central Nebraska before the 2007 and 2008 growing seasons. Six different strategies to predict OM were developed and tested by dividing samples randomly into calibration and validation datasets. Strategies included uniform, …

Soil Fertility And Crop Yields In Long-Term Organic And Conventional Cropping Systems In Eastern Nebraska, Samuel E. Wortman, Tomie D. Galusha, Stephen C. Mason, Charles A. Francis

Department of Agronomy and Horticulture: Faculty Publications

Organic agriculture aims to build soil quality and provide long-term benefits to people and the environment; however, organic practices may reduce crop yields. This long-term study near Mead, NE was conducted to determine differences in soil fertility and crop yields among conventional and organic cropping systems between 1996 and 2007. The conventional system (CR) consisted of corn (Zea mays L.) or sorghum (Sorghum bicolor (L.) Moench)–soybean (Glycine max (L.) Merr.)–sorghum or corn–soybean, whereas the diversified conventional system (DIR) consisted of corn or sorghum– sorghum or corn–soybean–winter wheat (wheat, Triticum aestivum L.). The animal manure-based organic system (OAM) …

Stacking Resistance Alleles From Wild And Domestic Soybean Sources Improves Soybean Cyst Nematode Resistance, Myungsik Kim, D. L. Hyten, Terry L. Niblack, Brian W. Diers

Department of Agronomy and Horticulture: Faculty Publications

The soybean cyst nematode (SCN; Heterodera glycines Ichinohe) is the most economically important soybean [Glycine max (L.) Merr.] pathogen in the United States. Field SCN populations are adapting to the narrowly based SCN resistance currently deployed in soybean cultivars. The objective of our research was to measure the effects of combinations of SCN resistance genes or quantitative trait loci (QTL) from the wild soybean (Glycine soja Siebold & Zucc.) PI 468916 and the domesticated soybean accessions PI 88788 and PI 437654. Two populations were developed to test the combinations of QTL and genes. Both populations segregated for the …

Moving Micronutrients From The Soil To The Seeds: Genes And Physiological Processes From A Biofortification Perspective, Brian M. Waters, Renuka P. Sankaran

Department of Agronomy and Horticulture: Faculty Publications

The micronutrients iron (Fe), zinc (Zn), and copper (Cu) are essential for plants and the humans and animals that consume plants. Increasing the micronutrient density of staple crops, or biofortification, will greatly improve human nutrition on a global scale. This review discusses the processes and genes needed to translocate micronutrients through the plant to the developing seeds, and potential strategies for developing biofortified crops.

2011 Wildflower Of The Year: White Oak, Quercus Alba, W. John Hayden

Biology Faculty Publications

Emblematic of strength and longevity, white oaks grace the deciduous forests of eastern North America.

Restoration Of Tall Fescue Pastures To Native Warm Season Grasslands: Does A Fungal Endophyte Symbiosis Play A Role In Restoration Success?, Sarah Lynn Hall

University of Kentucky Doctoral Dissertations

Tall fescue, a cool-season grass native to Europe, central Asia, and northern Africa, has been widely distributed throughout the U.S. for use as turf and forage. Following its widespread planting, its ability to associate with a toxic fungal endophyte, Neotyphodium coenophialum, was discovered. Research has linked this fescue-endophyte association with increased biotic and abiotic stress resistance in endophyte-infected (E+) versus endophyte-free (E-) plants, and these differences may affect the ability of land managers to eradicate tall fescue and restore native grasslands. I conducted three studies to examine whether E+ tall fescue plants respond differently to management than E- plants, …

T-Phylloplanin And Cis-Abienol, Two Natural Products From Tobacco Have Broad Spectrum, Anti-Fungal Activities, Brian Christopher King

University of Kentucky Doctoral Dissertations

Tobacco phylloplanins (T-phylloplanin) are a group of closely-related glycoproteins that are formed and disposed at the interface between the plant aerial surface (the phylloplane) and the atmosphere. They are synthesized in short procumbent trichomes and are secreted to aerial surfaces where they are thought to serve the plant as a first line of defense against fungal pathogens. Here it is shown using in vitro and in planta assays that tobacco and sunflower phylloplanins have broad-spectrum antifungal activities against spores - and also hyphae for two species - of several true fungi. Field tests show that T-phylloplanin reduces diseases caused by …

Can Resource-Use Traits Predict Native Vs. Exotic Plant Success In Carbon Amended Soils?, Robert J. Steers, Jennifer L. Funk, Edith B. Allen

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Productivity in desert ecosystems is primarily limited by water followed by nitrogen availability. In the deserts of southern California, nitrogen additions have increased invasive annual plant abundance. Similar findings from other ecosystems have led to a general acceptance that invasive plants, especially annual grasses, are nitrophilous. Consequently, reductions of soil nitrogen via carbon amendments have been conducted by many researchers in a variety of ecosystems in order to disproportionately lower invasive species abundance, but with mixed success. Recent studies suggest that resource-use traits may predict the efficacy of such resource manipulations; however, this theory remains largely untested. We report findings …

Elucidating The Biochemical Wizardry Of Triterpene Metabolism In Botroycoccus Braunii, Thomas Daniel Niehaus

Theses and Dissertations--Plant and Soil Sciences

B. braunii is a green alga that has attracted attention as a potential renewable fuel source due to its high oil content and the archeological record of its unique contribution to oil and coal shales. Three extant chemotypes of B. braunii have been described, namely race A, race B, and race L, which accumulate alkadienes and alkatrienes, botryococcene and squalene and their methylated derivatives, and lycopadiene, respectively. The methylated triterpenes, particularly botryococcenes, produced by race B can be efficiently converted to high quality combustible fuels and other petrochemicals; however, botryococcene biosynthesis has remained enigmatic.

It has been suggested that botryococcene …

Molecular And Chemical Dissection Of Cellulose Biosynthesis In Plants, Darby M. Harris

Theses and Dissertations--Plant and Soil Sciences

Plant cell walls are complex structures that must not only constrain cellular turgor pressure but also allow for structural modification during the dynamic processes of cell division and anisotropic expansion. Cell walls are composed of highly glycosylated proteins and polysaccharides, including pectin, hemicellulose and cellulose. The primary cell wall polysaccharide is cellulose, a polymer composed of high molecular weight !- 1,4-glucan chains. Although cellulose is the most abundant biopolymer on Earth, there is still a lot to learn about its biosynthesis and regulation. This research began by applying a variety of analytical techniques in an attempt to understand differences in …

Control Of Glucosylceramide Production And Morphogenesis By The Bar1 Ceramide Synthase In Fusarium Graminearum, William B. Rittenour, Ming Chen, Edgar B. Cahoon, Steven D. Harris

Department of Plant Pathology: Faculty Publications

The contribution of plasma membrane proteins to the virulence of plant pathogenic fungi is poorly understood. Accordingly, the objective of this study was to characterize the acyl-CoA dependent ceramide synthase Bar1 (previously implicated in plasma membrane organization) in the wheat pathogen Fusarium graminearum. The role of Bar1 in mediating cell membrane organization was confirmed as DBAR1 mutants failed to display a distinct sterol-rich domain at the hyphal tip. The DBAR1 mutants were non-pathogenic when inoculated onto wheat heads, and their in vitro growth also was severely perturbed. DBAR1 mutants were incapable of producing perithecia (sexual fruiting structures) and only produced …

The Sugar Sensor, Trehalose-6-Phosphate Synthase (Tps1), Regulates Primary And Secondary Metabolism During Infection By The Rice Blast Fungus: Will Magnaporthe Oryzae’S “Sweet Tooth” Become Its “Achilles’ Heel”?, Jesse Fernandez, Richard Wilson

Department of Plant Pathology: Faculty Publications

Rice blast disease is considered one of the most serious diseases of cultivated rice and is mediated by the causal agent, Magnaporthe oryzae. During infection, dome-shaped fungal cells, called appressoria, form on the surface of the leaf and generate turgor through the accumulation of glycerol. This enormous pressure is directed down onto a thin penetration hypha emerging from the base of the cell, forcing it through the surface of the rice leaf and allowing fungal colonization of the plant interior. The non-reducing disaccharide, trehalose, is present in conidia of M. oryzae and is mobilized during appressorium formation. The first …

Mechanisms Of Contact-Mediated Killing Of Yeast Cells On Dry Metallic Copper Surfaces, Davide Quaranta, Travis Krans, Christophe Espirito Santo, Christian Elowsky, Dylan W. Domaille, Christopher J. Chang, Gregor Grass

Department of Plant Pathology: Faculty Publications

Surfaces made of copper or its alloys have strong antimicrobial properties against a wide variety of microorganisms. However, the molecular mode of action responsible for the antimicrobial efficacy of metallic copper is not known. Here, we show that dry copper surfaces inactivate Candida albicans and Saccharomyces cerevisiae within minutes in a process called contact-mediated killing. Cellular copper ion homeostasis systems influenced the kinetics of contact-mediated killing in both organisms. Deregulated copper ion uptake through a hyperactive S. cerevisiae Ctr1p (ScCtr1p) copper uptake transporter in Saccharomyces resulted in faster inactivation of mutant cells than of wild-type cells. Similarly, lack of the …

Switchgrass Contains Two Cinnamyl Alcohol Dehydrogenases Involved In Lignin Formation, Aaron J. Saathoff, Christian M. Tobias, Scott E. Sattler, Eric J. Haas, Paul Twigg, Gautam Sarath

Department of Agronomy and Horticulture: Faculty Publications

Lignin content of switchgrass (Panicum virgatum L.), a bioenergy species, is a critical determinant of biomass quality since it can negatively impact conversion of biomass into liquid fuels via biochemical platforms. Cinnamyl alcohol dehydrogenase (CAD) is a key enzyme in lignin biosynthesis. Here, we have shown that cv. Kanlow switchgrass contains at least two closely related CAD genes (PviCAD1 and PviCAD2) that code for proteins containing highly conserved domains and residues that identify them as bona fide CADs. Both recombinant proteins displayed substrate kinetics consistent with their presumed role in cell wall lignification. Proteomic and immunoblotting detected …

The Composition And Origins Of Genomic Variation Among Individuals Of The Soybean Reference Cultivar Williams 82, William J. Haun, D. L. Hyten, Wayne W. Xu, Daniel J. Gerhardt, Thomas J. Albert, Todd Richmond, Jeffrey A. Jeddeloh, Gaofeng Jia, Nathan M. Springer, Carroll P. Vance, Robert M. Stupar

Department of Agronomy and Horticulture: Faculty Publications

Soybean (Glycine max) is a self-pollinating species that has relatively low nucleotide polymorphism rates compared with other crop species. Despite the low rate of nucleotide polymorphisms, a wide range of heritable phenotypic variation exists. There is even evidence for heritable phenotypic variation among individuals within some cultivars. Williams 82, the soybean cultivar used to produce the reference genome sequence, was derived from backcrossing a Phytophthora root rot resistance locus from the donor parent Kingwa into the recurrent parent Williams. To explore the genetic basis of intracultivar variation, we investigated the nucleotide, structural, and gene content variation of different …

Genome-Wide Association Analysis Identifies Candidate Genes Associated With Iron Deficiency Chlorosis In Soybean, Sujan Mamidi, Shireen Chikara, R. Jay Goos, D. L. Hyten, Deepti Annam, Samira Mafi Moghaddam, Rian K. Lee, P. B. Cregan, Phillip E. Mcclean

Department of Agronomy and Horticulture: Faculty Publications

Iron deficiency chlorosis (IDC) is a significant yield-limiting problem in several major soybean [Glycine max (L.) Merr.] production regions in the United States. Soybean plants display a variety of symptoms that range from a slight yellowing of the leaf to interveinal chlorosis, to stunted growth that reduces yield. The objective of this analysis was to employ single nucleotide polymorphism (SNP)-based genome-wide association mapping to uncover genomic regions associated with IDC tolerance. Two populations [2005 (n = 143) and 2006 (n = 141)] were evaluated in replicated, multilocation IDC trials. After controlling for population structure and individual relatedness, and selecting …